Boron-containing composition as well as preparation method and application thereof

Abstract

The invention discloses a boron-containing composition as well as a preparation method and application thereof. The invention provides a boron-containing composition. The boron-containing composition comprises 50-90 parts by weight of a matrix, 5-35 parts by weight of a boric acid compound and / or a derivative thereof and 1-30 parts by weight of an enzyme. The boron-containing composition provided by the invention can be used for preparing carbohydrates and derivatives, and particularly can improve the ratio of a product to a substrate, reduce the generation of byproducts and improve the product yield when being used for preparing micromolecular carbohydrates and derivatives. The boron-containing composition provided by the invention is insoluble, can be easily separated from a reaction liquid when being used as a catalyst for preparation of saccharides and derivatives, can be repeatedly used, is beneficial to reduction of later separation and purification cost, and has potential industrialization value.

Description

Technical Field

[0001] The present invention belongs to the technical field of sugar production, and relates to a boron-containing composition, a preparation method and use thereof, and specifically relates to a boron-containing composition and use thereof in the preparation of sugars and derivatives, and more specifically relates to a boron-containing composition catalyst and use thereof in the preparation of small molecule sugars and derivatives. Background Art

[0002] Carbohydrates are the most abundant organic compounds on earth and are widely present in biological cells. They play an important role in energy metabolism, maintaining cell integrity, molecular recognition, pathogen virulence, molecular defense, and signal information storage. Carbohydrates can be divided into monosaccharides, oligosaccharides, polysaccharides, and glycoconjugates according to their degree of polymerization. Monosaccharides are the simplest carbohydrates, which are single polyhydroxy aldehydes or polyhydroxy ketones. Most of the carbon atoms linked to hydroxyl groups in monosaccharides are chiral centers, thus producing many naturally occurring stereoisomers. Monosaccharides are divided into trisaccharides (including trisaccharides and acetose), tetrasaccharides (including butyrosinose and butyrosinose), pentoses (including pentoses and ketopentose), hexoses (including hexoses and hexoses), and heptoses according to the number of carbon atoms. Monosaccharides have a variety of applications in the pharmaceutical industry, biotechnology, and the food and beverage industry. Phosphate monosaccharides are widely present in various cells and are the main participants in many metabolic pathways. For example, D-glucose 1-phosphate, D-glucose 6-phosphate, D-fructose 6-phosphate and fructose 1,6-diphosphate are intermediates in the glycolysis pathway. Deoxy sugars are compounds in which one or more hydroxyl groups in a monosaccharide molecule are replaced by hydrogen atoms, and are widely distributed in plants, bacteria and animals. Disaccharides are the simplest oligosaccharides, which are formed by the condensation of two monosaccharides. Some disaccharides are widely present in nature, such as sucrose, maltose, cellobiose, trehalose, lactose, etc.

[0003] Monosaccharides and their derivatives can be prepared by isomerase catalysis (Susumu Ito, Features and applications of microbial sugar epimerases, Appl Microbiol Biotechnol (2009) 84: 1053–1060; Koen Beerens, Tom Desmet, Wim Soetaert, Enzymes for the biocatalytic production of rare sugars, J Ind Microbiol Biotechnol (2012) 39:823–834), for example, xylose isomerase catalyzes the conversion of D-glucose into D-fructose, L-arabinose isomerase catalyzes the conversion of D-galactose into D-tagatose, L-arabinose isomerase catalyzes the conversion of L-arabinose into L-ribulose, xylose isomerase catalyzes the conversion of D-xylose into D-xylulose, D-psicose 3-epimerase catalyzes the conversion of D-fructose into D-psicose, and fructose 4-epimerase catalyzes the conversion of D-fructose into D-tagatose. D-xylulose 4-epimerase catalyzes the conversion of D-xylulose to L-ribulose, D-glucose 6-phosphate isomerase catalyzes the conversion of D-glucose 6-phosphate to D-fructose 6-phosphate, D-psicose-6-phosphate 3-epimerase catalyzes the conversion of D-fructose 6-phosphate to D-psicose 6-phosphate, D-tagatose 6-phosphate 4-epimerase catalyzes the conversion of D-fructose 6-phosphate to D-tagatose 6-phosphate, and D-tagatose 3-epimerase catalyzes the epimerization of 6-deoxy-L-fructose to 6-deoxy-L-psicose, etc. Disaccharides can also be prepared by isomerase catalysis, for example, cellobiose epimerase catalyzes the conversion of lactose to lactulose. However, the preparation of carbohydrate compounds using isomerases generally has the problem that most substrates cannot be converted into products. Therefore, it is necessary to develop a new strategy for producing carbohydrate compounds, especially small molecule sugars, in a manner with a high product to substrate ratio. Summary of the invention

[0004] Problem that the invention aims to solve

[0005] In view of the above problems, the inventors of the present invention have conducted in-depth research and successfully developed a novel boron-containing composition, which can improve the ratio of product to substrate when used to prepare sugars (e.g., monosaccharides, disaccharides, polysaccharides) and derivatives (e.g., phosphate sugars, deoxy sugars), especially small molecule sugars (e.g., monosaccharides, disaccharides) and derivatives (e.g., phosphate sugars, deoxy sugars).

[0006] Solutions for solving problems

[0007] [1]. A boron-containing composition comprising 50-90 parts by weight of a substrate, 5-35 parts by weight of a boric acid compound and / or its derivatives and 1-30 parts by weight of an enzyme.

[0008] [2] The boron-containing composition according to [1], wherein the boron-containing composition is used as a catalyst;

[0009] Preferably, the boron-containing composition is used as a catalyst for the production of sugars and / or their derivatives.

[0010] [3] The boron-containing composition according to [2], wherein the sugar comprises one or more of a monosaccharide, a disaccharide, an oligosaccharide and a polysaccharide;

[0011] Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides;

[0012] More preferably, the sugar includes one or more of monosaccharides and disaccharides.

[0013] [4] The boron-containing composition according to [2] or [3], wherein the sugar derivative comprises one or more of phosphate sugar and deoxy sugar.

[0014] [5] A boron-containing composition according to any one of [1] to [4], wherein the matrix comprises any substance capable of loading a boric acid compound and / or its derivatives and / or an enzyme, preferably, the matrix comprises one or more of an organic synthetic polymer, an inorganic substance, a natural organic macromolecular compound, a magnetic particle, a metal organic framework material, a covalent organic framework material, an organosiloxane polymer layer and a macroporous resin.

[0015] [6] The boron-containing composition according to any one of [1] to [5], wherein the boric acid compound and / or its derivative is supported on the substrate;

[0016] Optionally, the enzyme is supported on the matrix.

[0017] [7] The boron-containing composition according to any one of [1] to [6], wherein

[0018] The boronic acid compound and / or its derivative and the enzyme are respectively loaded on the same substrate; and / or,

[0019] The boronic acid compound and / or its derivative and the enzyme are respectively loaded on different substrates; and / or,

[0020] The boronic acid compound and / or its derivative and the enzyme are simultaneously loaded on the same substrate; and / or,

[0021] The boric acid compound and / or its derivative is loaded on the matrix, and the enzyme is in a free form;

[0022] Optionally, the loading is achieved by adsorption, embedding, cross-linking, covalent binding or a combination thereof.

[0023] [8] The boron-containing composition according to any one of [1] to [7], wherein the boron-containing composition further comprises a linker to load the boronic acid compound and / or its derivative and / or the enzyme on the substrate;

[0024] Optionally, the linker is a cross-linker.

[0025] [9] The boron-containing composition according to any one of [1] to [8], wherein the enzyme comprises one or more enzymes from the isomerase group belonging to EC number EC5;

[0026] Optionally, the enzyme comprises one or more enzymes from the group of glycosyltransferases belonging to EC numbering EC 2.4.

[0027]

[10] The boron-containing composition according to any one of [1] to [9], wherein the enzyme comprises one, two, three or more enzymes having the same or different functions.

[0028]

[11] . The boron-containing composition according to any one of [1] to

[10] , wherein the boric acid compound and / or its derivative comprises one or more of boric acid, inorganic borates, organic boric acid and organic borates.

[0029]

[12] A boron-containing composition according to any one of [1] to

[11] , wherein the boron-containing composition is in the form of one or more of beads, spheres, particles, granules, gels, fibers, films, or surfaces.

[0030]

[13] A method for preparing a boron-containing composition as described in any one of [1] to

[12] , the method comprising the steps of mixing a boric acid compound and / or its derivative and / or an enzyme with a substrate, loading the substrate, and obtaining the boron-containing composition.

[0031]

[14] Any of the following uses of the boron-containing composition according to any one of [1] to

[12] or the boron-containing composition prepared by the preparation method according to

[13] :

[0032] (a) Use in the preparation of sugars and / or their derivatives;

[0033] (b) Use as a catalyst for preparing sugars and / or their derivatives;

[0034] Optionally, the sugar comprises one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides; the sugar derivative comprises one or more of phosphate sugars and deoxy sugars;

[0035] Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides;

[0036] More preferably, the sugar includes one or more of monosaccharides and disaccharides.

[0037]

[15] . A method for preparing sugar and / or its derivatives, comprising the steps of:

[0038] (i) preparing a reaction mixture comprising a substrate,

[0039] (ii) contacting the reaction mixture with a boron-containing composition under suitable reaction conditions;

[0040] The boron-containing composition is a boron-containing composition according to any one of [1] to

[12] or a boron-containing composition prepared by the preparation method according to

[13] .

[0041]

[16] The method according to

[15] , wherein the substrate comprises sugar and / or its derivatives;

[0042] Optionally, the sugar comprises one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides; the sugar derivative comprises one or more of phosphate sugars and deoxy sugars;

[0043] Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides;

[0044] More preferably, the sugar includes one or more of monosaccharides and disaccharides.

[0045]

[17] The use or method according to any one of

[14] to

[16] , wherein the boron-containing composition is used to prepare sugar and / or its derivatives under acidic conditions, neutral conditions or alkaline conditions,

[0046] Optionally, the acidic condition is a pH value lower than 7.0, the neutral condition is a pH value equal to 7.0, and the alkaline condition is a pH value higher than 7.0.

[0047]

[18] The use or method according to any one of

[14] to

[17] , wherein the boron-containing composition is used in solution to prepare sugar and / or its derivatives,

[0048] Optionally, the solution is selected from an aqueous solution, an organic solvent, an ionic liquid or a deep eutectic solvent, and / or,

[0049] The solution is selected from a buffer or a non-buffer;

[0050] Preferably, the buffer is selected from acetate buffer, citrate buffer, glycine buffer, phthalate buffer, phosphate buffer, 4-hydroxyethylpiperazineethanesulfonic acid buffer, 3-morpholinepropanesulfonic acid buffer, tris buffer, carbonate buffer or ammonia-ammonium chloride buffer.

[0051] Effects of the Invention

[0052] 1. The boron-containing composition of the present invention is used for the preparation of sugars and derivatives, especially small molecule sugars and derivatives, to increase the ratio of product to substrate, reduce the generation of by-products, and increase the product yield.

[0053] 2. The boron-containing composition of the present invention is used for the preparation of sugars and derivatives, especially for the preparation of monosaccharides and derivatives via phosphate monosaccharide compounds, which can reduce the amount of by-products generated and improve the yield of the product.

[0054] 3. The boric acid compound and / or its derivative in the boron-containing composition of the present invention is loaded on a matrix and is insoluble. When used for the preparation of sugars and derivatives, the catalyst and the product are easily separated from the reaction solution, which is beneficial to reduce the cost of subsequent separation and purification and has potential industrial value. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 Four exemplary forms of the boron-containing composition of the present invention are shown, wherein E (Enzyme) represents an enzyme, including a purified or partially purified enzyme, a microorganism expressing the enzyme, a culture of the microorganism, or a combination thereof.

[0056] Figure 2 The conversion between pentose sugars and their derivatives catalyzed by isomerase is shown.

[0057] Figure 3 The conversion between hexoses and derivatives catalyzed by isomerases is shown.

[0058] Figure 4 The HPLC test result graph in Example 1 is shown.

[0059] Figure 5 The HPLC test result graph in Example 2 is shown.

[0060] Figure 6 The HPLC detection result graph in Example 4 is shown. DETAILED DESCRIPTION

[0061] Various exemplary embodiments, features and aspects of the present invention will be described in detail below. The word "exemplary" used here means "used as an example, embodiment or illustrative". Any embodiment described here as "exemplary" is not necessarily interpreted as being superior or better than other embodiments.

[0062] In addition, in order to better illustrate the present invention, numerous specific details are provided in the following specific embodiments. It should be understood by those skilled in the art that the present invention can be implemented without certain specific details. In other examples, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the subject matter of the present invention.

[0063] Unless otherwise stated, the units used in this specification are all international standard units, and the numerical values ​​and numerical ranges appearing in the present invention should be understood to include the inevitable systematic errors in industrial production.

[0064] In this specification, the word "may" means both performing a certain process and not performing a certain process.

[0065] In this specification, the references to "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc., mean that the specific elements (e.g., features, structures, properties and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not exist in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0066] In this specification, the numerical range expressed using "a numerical value A to a numerical value B" means a range including the endpoints numerical values ​​A and B.

[0067] In this specification, when "normal temperature" or "room temperature" is used, the temperature may be 10-40°C.

[0068] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used in the text have the common meanings understood by those skilled in the art for the present invention. In the event of a conflict, the definitions in this specification shall prevail.

[0069] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0070] In the present invention, unless otherwise specified, all embodiments and preferred embodiments mentioned herein can be combined with each other to form a new technical solution.

[0071] In the present invention, unless otherwise specified, all technical features and preferred features mentioned herein can be combined with each other to form a new technical solution.

[0072] The use of "one", "a", "an" or similar expressions to describe the components and technical features of the present invention is merely for the convenience of expression and to provide a general meaning to the scope of the present invention. Therefore, such description should be understood to include one or at least one, and the singular also includes the plural, unless it is obvious that it refers to another meaning.

[0073] In the present invention, "multiple" and "multiple" refer to two (two kinds) or more than two (two kinds). "And / or" describes the association relationship of the associated objects, indicating that three relationships may exist. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. The character " / " generally indicates that the associated objects before and after are in an "or" relationship.

[0074] In this document, "or its combination" means "or any combination thereof", and "any one", "any one" means "any one", "any one" or "any one".

[0075] The terms "preferably" and "preferred" as used herein do not limit the scope of the claimed invention, or imply that certain features are critical, necessary, or even important to the structure or function of the claimed invention. Instead, these terms are merely intended to emphasize alternative or additional features that can or cannot be used in a particular embodiment of the invention.

[0076] In the present invention, if there is no special explanation, the "including" mentioned herein means open or closed. For example, the "including" may mean that other components not listed may also be included, or only the listed components may be included.

[0077] As used herein, the term "derivative" refers to a compound having a structure derived from a parent compound, whose structure is sufficiently similar to those disclosed herein, and based on the similarity, a person skilled in the art would expect to have the same or similar activity and utility as the claimed compound. Exemplary derivatives include salts, hydrates, phosphate compounds, deoxy compounds, etc. of the parent compound.

[0078] As used herein, the term "matrix" refers to an inert, functionalized, reusable, insoluble medium that provides physical stability (e.g., mechanical or binding stability). Boric acid compounds and / or derivatives or enzymes can be attached to them directly or via a connector (carrying an anchoring group) to form a boron-containing composition or boron-containing composition catalyst of the present invention. When the boron-containing composition or boron-containing composition catalyst is used for reaction, the boron-containing composition or boron-containing composition catalyst can be easily separated from excess reagents, soluble reaction products, by-products or solvents (by washing, filtering, centrifugation, etc.).

[0079] As used herein, the term "loading" refers to a process of combining a boric acid compound and / or its derivative or enzyme with a substrate. After the boric acid compound and / or its derivative or enzyme is combined with the substrate to form the boron-containing composition or boron-containing composition catalyst of the present invention, the boron-containing composition or boron-containing composition catalyst will exist in the form of insoluble matter.

[0080] In the present invention, the term "saccharide" or "sugar" is intended to cover all carbohydrates (sugars), including naturally occurring and synthetic and semi-synthetic carbohydrates. The term covers monosaccharides (i.e., carbohydrates that cannot be hydrolyzed into simpler sugars), disaccharides (i.e., compounds having two monosaccharide units (parts) linked together by glycosidic bonds), oligosaccharides (i.e., compounds having 3 to 10 monosaccharide units linked together by glycosidic bonds, in branched or unbranched chains or rings (optionally with carbohydrate side chains)), and polysaccharides (i.e., compounds having more than 10 monosaccharide units linked together by glycosidic bonds, in branched or unbranched chains or rings (optionally with carbohydrate side chains)).

[0081] In the present invention, the term "small molecule sugar" refers to sugars having a degree of polymerization DP of less than 10. In particular, in the present invention, small molecule sugars include monosaccharides and disaccharides.

[0082] In the present invention, the term "monosaccharide" refers to a sugar that cannot be hydrolyzed into simpler sugars. The monosaccharide can have a D- or L- configuration and can be an aldose or a ketose. Examples of monosaccharides include pentoses and hexoses. Pentose includes aldopentose (referring to a sugar containing 5 carbon atoms and containing an aldehyde functional group) and ketopentose (referring to a sugar containing 5 carbon atoms and containing a keto functional group). Examples of pentoses include Figure 2 Those listed, for example, ribose, arabinose, xylose, lyxose, ribulose, xylulose. Hexoses include aldohexoses (referring to sugars containing 6 carbon atoms and containing an aldehyde functional group) and ketohexoses (referring to sugars containing 6 carbon atoms and containing a keto functional group). Examples of hexoses are Figure 3 Those listed, for example, allose, altrose, glucose, mannose, gulose, idose, galactose, talose, psicose, fructose, sorbose, tagatose.

[0083] In the present invention, the term "monosaccharide phosphate", "monosaccharide phosphate" or "phosphorylated monosaccharide" refers to a compound formed by replacing one or more hydroxyl groups in a monosaccharide with a phosphate group. In particular, it refers to a compound formed by replacing the hydroxyl group at position 6 in a monosaccharide with a phosphate group. Some examples of monosaccharides phosphate include Figure 3 and Figure 4 Those listed, for example, allose 6-phosphate, altrose 6-phosphate, glucose 6-phosphate, mannose 6-phosphate, gulose 6-phosphate, idose 6-phosphate, galactose 6-phosphate, talose 6-phosphate, psicose 6-phosphate, fructose 6-phosphate, sorbose 6-phosphate, tagatose 6-phosphate, ribose 5-phosphate, ribulose 5-phosphate, xylulose 5-phosphate.

[0084] In the present invention, the term "deoxy sugar" refers to a compound formed when one or more hydroxyl groups of a monosaccharide molecule are replaced by a hydrogen atom. In particular, it refers to a compound formed when the hydroxyl group at position 1 or 6 of a monosaccharide is replaced by a hydrogen atom. Some examples of 6-deoxy sugars include Figure 3 and Figure 4 Those listed. Examples of 1-deoxy sugars include 1-deoxy-L-fructose, 1-deoxy-L-psicose, 1-deoxy-L-tagatose, 1-deoxy-L-sorbose. Examples of 6-deoxy sugars include 6-deoxy-L-mannose (L-rhamnose), 6-deoxy-L-galactose (L-fucose), 6-deoxy-D-galactose (D-fucose), 6-deoxy-D-glucose (iso-rhamnose).

[0085] In the present invention, the term "disaccharide" or "disaccharide" refers to a compound in which two monosaccharide units are linked by a glycosidic bond. Examples of disaccharides include sucrose, maltose, cellobiose, lactose, lactulose, trehalose, turanoose, trehalulose, isomaltose, isomaltulose, kojibiose, nigerose, sophorose, laminaribiose, gentiobiose, melibiose, melibiose (melbiose), allolactose, rutinose, rutinulose, acaciabiose, and neohesperidose.

[0086] As used herein, the term "suitable reaction conditions" refers to those conditions in a reaction solution catalyzed by a boron-containing composition or a boron-containing composition catalyst, such as ranges of catalyst loading, substrate loading, temperature, pH, buffer, metal ions, etc. Some exemplary "suitable reaction conditions" are provided herein.

[0087] EC number or EC number is a classification system for enzymes created by the Enzyme Commission, based on the chemical reaction catalyzed by each enzyme. This classification system also gives a suggested name to each enzyme, so it is also called the Enzyme Commission Nomenclature.

[0088] <Boron-containing composition>

[0089] A first aspect of the present invention provides a boron-containing composition, the boron-containing composition comprising:

[0090] (a) 50-90 parts by weight of a matrix;

[0091] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0092] (c) 1-30 parts by weight of an enzyme.

[0093] Preferably, the boron composition is used as a catalyst. Therefore, the present invention provides a boron-containing composition catalyst, which comprises:

[0094] (a) 50-90 parts by weight of a matrix;

[0095] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0096] (c) 1-30 parts by weight of an enzyme.

[0097] More preferably, the boron composition is used as a catalyst for producing sugars and / or their derivatives. Therefore, the present invention provides a boron-containing composition catalyst for producing sugars (e.g., monosaccharides, disaccharides, polysaccharides) and derivatives (e.g., phosphate sugars, deoxy sugars), the boron-containing composition catalyst comprising:

[0098] (a) 50-90 parts by weight of a matrix;

[0099] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0100] (c) 1-30 parts by weight of an enzyme.

[0101] Further preferably, the present invention provides a boron-containing composition catalyst for producing small molecule sugars (e.g., monosaccharides, disaccharides) and derivatives (e.g., phosphate sugars, deoxy sugars), the boron-containing composition catalyst comprising:

[0102] (a) 50-90 parts by weight of a matrix;

[0103] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0104] (c) 1-30 parts by weight of an enzyme.

[0105] More preferably, the present invention provides a boron-containing composition catalyst for increasing the ratio of product to substrate when producing small molecule sugars (e.g., monosaccharides, disaccharides) and derivatives (e.g., phosphate sugars, deoxy sugars), the boron-containing composition catalyst comprising:

[0106] (a) 50-90 parts by weight of a matrix;

[0107] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0108] (c) 1-30 parts by weight of an enzyme.

[0109] In some embodiments of the present invention, the sugar comprises one or more of a monosaccharide, a disaccharide, an oligosaccharide, and a polysaccharide.

[0110] In some preferred embodiments of the present invention, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides.

[0111] In some more preferred embodiments of the present invention, the sugar includes one or more of monosaccharides and disaccharides.

[0112] In some embodiments of the present invention, the sugar derivatives include one or more of phosphate sugars and deoxy sugars.

[0113] In some exemplary embodiments, the monosaccharide includes fructose, xylulose, psicose, tagatose, and ribulose, but is not limited thereto.

[0114] In some exemplary embodiments, the phosphate sugar is a phosphate monosaccharide, which includes fructose 6-phosphate and psicose 6-phosphate, but is not limited thereto.

[0115] In some exemplary embodiments, the deoxysugar includes L-rhamnulose (6-deoxyL-fructose), 6-deoxyL-tagatose, 6-deoxyL-psicose, 1-deoxyL-psicose, but is not limited thereto.

[0116] In some exemplary embodiments, the disaccharide includes turanose and lactulose, but is not limited thereto.

[0117] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing lactulose and / or its derivatives, the boron-containing composition catalyst comprising:

[0118] (a) 50-90 parts by weight of a matrix;

[0119] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0120] (c) 1-30 parts by weight of an enzyme.

[0121] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing fructose and / or its derivatives, the boron-containing composition catalyst comprising:

[0122] (a) 50-90 parts by weight of a matrix;

[0123] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0124] (c) 1-30 parts by weight of an enzyme.

[0125] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing psicose and / or its derivatives, wherein the boron-containing composition catalyst comprises:

[0126] (a) 50-90 parts by weight of a matrix;

[0127] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0128] (c) 1-30 parts by weight of an enzyme.

[0129] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing xylulose and / or its derivatives, the boron-containing composition catalyst comprising:

[0130] (a) 50-90 parts by weight of a matrix;

[0131] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0132] (c) 1-30 parts by weight of an enzyme.

[0133] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing tagatose and / or its derivatives, the boron-containing composition catalyst comprising:

[0134] (a) 50-90 parts by weight of a matrix;

[0135] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0136] (c) 1-30 parts by weight of an enzyme.

[0137] In some exemplary embodiments, the present invention provides a boron-containing composition catalyst for producing turanoose and / or its derivatives, the boron-containing composition catalyst comprising:

[0138] (a) 50-90 parts by weight of a matrix;

[0139] (b) 5-35 parts by weight of a boric acid compound and / or a derivative thereof;

[0140] (c) 1-30 parts by weight of an enzyme.

[0141] In some optional embodiments, the boron-containing composition / boron-containing composition catalyst includes 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90 (or any value between any adjacent values) parts by weight of the matrix.

[0142] In some optional embodiments, the boron-containing composition / boron-containing composition catalyst includes 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 (or any value between any adjacent values) parts by weight of a boric acid compound and / or its derivative.

[0143] In some optional embodiments, the boron-containing composition / boron-containing composition catalyst includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 (or any value between any adjacent values) weight parts of enzyme.

[0144] It should be noted that the enzyme used in the boron-containing composition / boron-containing composition catalyst of the present invention can increase or reduce the enzyme activity unit used according to actual needs, so that the enzyme remains within the weight range of the present invention. Simple conversion can be carried out according to actual demand between the activity unit and the weight of the enzyme. For example, the activity of the corresponding functional enzyme used is low, and in order to reduce the reaction time and keep the reaction rate, the consumption of the component enzyme can be increased. Those skilled in the art can select its concrete enzyme activity unit according to actual needs, within the weight range of enzyme of the present invention.

[0145] In some embodiments, the component (b) boronic acid compound and / or its derivative is supported on the component (a) matrix. Optionally, the component (c) enzyme is supported on the component (a) matrix.

[0146] In some embodiments of the present invention, the boron-containing composition or boron-containing composition catalyst may be in the form of beads, spheres, particles, granules, gels, fibers, films, or surfaces, but is not limited thereto.

[0147] Component (a) Matrix

[0148] In the present invention, component (a) matrix can be any material that can load component (b) boric acid compound and / or its derivative and / or component (c) enzyme. In some embodiments of the present invention, described component (a) matrix can be organic synthetic polymer, for example polystyrene, polyethylene, polypropylene, polyfluoroethylene, polyoxyethylene, polyacrylamide, polyurethane, polycarbonate, polyacrylic acid, polyacrylate, polyvinyl pyrrolidone, polyethylene glycol, polyvinyl alcohol, N-(2-hydroxypropyl)-methacrylamide, polydimethylsiloxane and its copolymer and graft or one or more of its combination. These organic synthetic polymers can also be functionalized with epoxy, amino or alkyl groups, and these functionalized polymers are also included in component (a) matrix of the present invention. In some embodiments of the present invention, the matrix of component (a) may be an inorganic substance, such as diatomaceous earth, bentonite, kaolin, montmorillonite, expanded perlite, silica gel, glass (e.g., controlled porosity glass, high boron glass), ceramics, zeolite, silicon oxide, silicon dioxide, metal or carbon-based materials (e.g., activated carbon, charcoal, graphene), etc. In some embodiments of the present invention, the matrix of component (a) may be a natural organic macromolecular compound, such as alginate (e.g., calcium alginate), chitosan, collagen, carrageenan, gelatin, cellulose, starch, pectin or agarose, etc. In some embodiments of the present invention, the matrix of component (a) may include or be composed of magnetic particles, such as magnetic nanoparticles, magnetic ferrosoferric oxide. In some embodiments of the present invention, the matrix of component (a) may be a metal organic framework (MOF) material, such as zeolite imidazolate framework-90 (ZIF-90). In some embodiments of the present invention, the component (a) matrix can be a covalent organic framework (COF) material, such as a vinyl-linked covalent organic framework material, an imide-linked covalent organic framework material, a borate-linked covalent organic framework material, a boron-oxygen six-membered ring-linked covalent organic framework material, an imine-linked covalent organic framework material, an azine-linked covalent organic framework material, a keto-enol-linked covalent organic framework material, a hydrazone-linked covalent organic framework material, or a triazine-linked covalent organic framework material. In some embodiments of the present invention, the component (a) matrix can be an organosiloxane polymer layer, such as a (3-aminopropyl) trimethoxysilane (APTMS) monomer, a trimethoxy (propyl) silane (TMPS) monomer, or a combination thereof. In some specific embodiments of the present invention, the component (a) matrix can be a resin (e.g., a macroporous resin), such as an epoxy resin (epoxy macroporous resin) or an amino resin (amino macroporous resin).

[0149] In the present invention, component (a) substrate can be in various forms, such as beads, balls, particles, granules, gels, fibers, membranes (such as hollow fiber membranes, mixed matrix membranes) or surfaces. The surface can be planar, substantially planar or non-planar. In the present invention, component (a) substrate can be porous or non-porous, and can have expansion or non-expansion characteristics. For example, component (a) substrate can be a porous carbon material, a porous silica material.

[0150] Component (b) Boric acid compound and / or its derivative

[0151] In the present invention, component (b) boric acid compound and / or its derivatives include but are not limited to boric acid (H 3 BO 3 ), inorganic borates, organic boric acids and / or organic borates.

[0152] In the present invention, the boric acid (H 3 BO 3 ), inorganic borates, organic boronic acids and / or organic borates may be used as commercially available products containing the above substances.

[0153] In some embodiments of the present invention, the inorganic borate may be a salt in which the hydrogen atom of boric acid is replaced by a metal element, for example, sodium borate, magnesium borate, zinc borate, calcium borate, barium borate. Sodium borate is a general term for metal salts containing boron and sodium bonded to oxygen, for example, anhydrous borax (Na 2 B 4 O 7 ), borax pentahydrate (borax trihydrate, Na 2 B 4 O 7 ·5H 2 O), borax decahydrate (Na 2 B 4 O 7 10H 2 O), etc. Magnesium borate is a general term for metal salts containing boron and magnesium bonded to oxygen, for example, magnesium orthoborate (Mg 3 (BO 3 ) 2 ), magnesium diborate (magnesium pyroborate, Mg 2 B 2 O 5 or 2MgO·B 2 O 3 ), magnesium metaborate (MgO·B 2 O 3 ), trimagnesium tetraborate (Mg 3 B 4 O 9 or 3MgO·2B 2 O 3), pentamagnesium tetraborate (Mg 5 B 4 O 11 ), magnesium hexaborate (MgB 6 O 10 ), Mg 2 [B 2 O 4 (OH)](OH) etc. In addition, examples of magnesium borate include 2MgO·3B 2 O 3 ·nH 2 O (where n is a positive integer), MgO·4B 2 O 3 ·3H 2 O or MgO·6B 2 O 3 18H 2 O etc., but are not limited thereto. Zinc borate is a general term for metal salts containing boron and zinc bonded to oxygen, such as zinc orthoborate (Zn 3 (BO 3 ) 2 )、Zn(BO 2 ) 2 ), basic zinc borate (ZnB 4 O 7 ·2ZnO), 3.5 hydrated zinc borate (2ZnO·3B 2 O 3 3.5H 2 O), zinc borate heptahydrate (2ZnO·3B 2 O 3 7H 2 O), zinc borate pentahydrate (2ZnO·3B 2 O 3 ·5H 2 O), zinc borate dihydrate (ZnO·B 2 O 3 ·2H 2 O), etc. Calcium borate is a general term for metal salts containing boron and calcium bonded to oxygen, such as calcium tetraborate, calcium metaborate, etc. Barium borate is a general term for metal salts containing boron and barium bonded to oxygen, such as barium borate monohydrate, barium metaborate, etc.

[0154] In some embodiments of the present invention, the organic boronic acid corresponds to formula (I): RB(OH) 2 ,

[0155] Wherein R is selected from alkyl, alkenyl, aryl or heteroaryl, which may or may not be substituted by at least one substituent selected from the following: halogen group, substituted alkyl or alkenyl group, oxo group, amino and substituted amino group, nitro group, aldehyde group, carboxylic acid and ester, ketone, sulfone, sulfonic acid group.

[0156] In some specific embodiments, when the R group is an alkyl or alkenyl group containing 1-15 carbon atoms, the R group preferably contains 1-10 carbon atoms, more preferably 1-6 carbon atoms.

[0157] In other specific embodiments, when the R group is a cycloalkyl group containing 3 to 6 carbon atoms, the R group is preferably selected from cyclohexyl or cyclopentyl.

[0158] In some specific embodiments, when the R group is an aryl group containing 5 to 15 carbon atoms, the R group preferably contains 5 to 10 carbon atoms, more preferably 5 to 6 carbon atoms. Further preferably, the group R is selected from phenyl and naphthyl.

[0159] In other specific embodiments, when the R group is an aryl group substituted by one or more substituents, the substituents are selected from alkenyl, amino, carboxyl, and acyl.

[0160] In some preferred embodiments, the organic boronic acid can be selected from cyclohexylboronic acid, phenylboronic acid, naphthylboronic acid, aminophenylboronic acid (2-aminophenylboronic acid, 3-aminophenylboronic acid, 4-aminophenylboronic acid), carboxyphenylboronic acid (2-carboxyphenylboronic acid, 3-carboxyphenylboronic acid, 4-carboxyphenylboronic acid), 3-formylphenylboronic acid, 3-acrylamidophenylboronic acid, 4-vinylphenylboronic acid, etc.

[0161] In some embodiments of the present invention, the organic borate may be selected from aminophenyl borate (2-aminophenylborate hydrochloride, 3-aminophenylboronic acid hemisulfate, 3-aminophenylborate hydrochloride), carboxyphenyl borate (sodium 2-carboxyphenylborate), and the like.

[0162] Component (c) Enzyme

[0163] In the present invention, the component (c) enzyme includes an enzyme that catalyzes an isomerization reaction, specifically including any one or more of all enzymes belonging to the "EC5 (isomerase)" group in the EC number. In some embodiments, the component (c) enzyme also includes any one or more of the enzymes from the EC number belonging to the EC2.4 glycosyltransferase group.

[0164] In some optional embodiments, the component (c) enzyme includes one or more of all enzymes of the EC5.1 (racemases and epimerases), EC5.2 (cis-trans isomerases), EC5.3 (intramolecular isomerases), EC5.4 (mutases), EC5.5 (intramolecular lyases), and EC5.6 (other isomerases) groups.

[0165] In some preferred embodiments, isomerase refers to any enzyme that can convert a sugar (e.g., monosaccharide, disaccharide, polysaccharide) and / or its derivative (e.g., phosphate sugar, deoxy sugar) molecule from one isomer to another isomer. Preferably, isomerase refers to any enzyme that can convert a small molecule sugar (e.g., monosaccharide or disaccharide) and derivative (e.g., phosphate monosaccharide or deoxy sugar) molecule from one isomer to another isomer. In the present invention, epimerase refers to an enzyme that moves the position of -OH bonded to C in a monosaccharide molecule to the opposite side. Therefore, epimerase is an enzyme belonging to isomerase.

[0166] Isomerases can promote intramolecular rearrangements, in which bonds are broken and formed, or they can catalyze conformational changes. For example, aldose-ketose isomerase catalyzes the movement of hydrogen within the molecule, promotes the redox reaction between C1 and C2, and catalyzes the mutual conversion of aldose and ketose. All proteins that have isomerase activity in organisms such as microorganisms, plants, and animals are also included. Even newly discovered enzymes and enzymes classified as other isomerases are included as long as they have an isomerization effect, especially a catalytic effect of isomerizing small molecule sugars (e.g., monosaccharides or disaccharides) and derivatives (e.g., phosphate monosaccharides or deoxy sugars).

[0167] The component (c) enzyme that can be used in the present invention is a wild-type enzyme or an enzyme that has undergone certain genetic modifications, and the genetic modifications have improved some properties of the isomerase, such as activity, substrate specificity, thermal stability, etc. Enzymes with desired properties derived from wild-type enzymes by genetic engineering techniques are called "mutants", "mutant derivatives" or "mutants from the corresponding wild-type enzymes". In the present invention, the isomerase used to implement the present invention includes "mutants" with improved properties.

[0168] The substrates suitable for the enzyme of component (c) of the present invention include, but are not limited to, sugars, phosphate sugars, and deoxy sugars. Preferably, the substrates of the enzyme of component (c) are monosaccharides, disaccharides, phosphate monosaccharides, 6-deoxy sugars, and 1-deoxy sugars.

[0169] In some embodiments of the present invention, xylose isomerase (EC5.3.1.5) refers to an enzyme that catalyzes the reversible isomerization of 3S / 4R ketose to 2R / 3S / 4R aldose and the reversible isomerization of 3R / 4S ketose to 2S / 3R / 4S aldose. Isomerization reactions catalyzed by xylose isomerase include but are not limited to the isomerization of D-glucose to D-fructose, the isomerization of D-xylose to D-xylulose, the isomerization of D-sorbose to D-idose and the isomerization of L-sorbose to L-idose. In the present invention, the above-mentioned "enzyme that isomerizes D-glucose to D-fructose" includes, for example, L-rhamnose isomerase of a certain microorganism that acts on D-glucose and converts it into D-fructose. The case where proteins contained in organisms such as all microorganisms, plants, and animals have the function of converting D-glucose into D-fructose is also included. The isomerization of other sugars and / or their derivatives has the same meaning.

[0170] In some embodiments of the present invention, mannose isomerase (EC5.3.1.7) refers to an enzyme that catalyzes the reversible isomerization of 3S / 4R ketose to 2S / 3S / 4R aldose. Isomerization reactions catalyzed by mannose isomerase include, but are not limited to, the isomerization of D-mannose to D-fructose, the isomerization of D-xylulose to D-lyxose, and the isomerization of L-sorbose to L-gulose.

[0171] In some embodiments of the present invention, L-arabinose isomerase (EC5.3.1.4) refers to an enzyme that catalyzes the reversible isomerization of 3S / 4S ketose to 2R / 3S / 4S aldose. Isomerization reactions catalyzed by L-arabinose isomerase include, but are not limited to, the isomerization of D-tagatose to D-galactose, the isomerization of L-psicose to L-altrose, and the isomerization of L-ribulose to L-arabinose.

[0172] In some embodiments of the present invention, D-arabinose isomerase (EC5.3.1.3) refers to an enzyme that catalyzes the reversible isomerization of 3R / 4R ketose to 2S / 3R / 4R aldose. Isomerization reactions catalyzed by D-arabinose isomerase include but are not limited to the isomerization of D-psicose to D-altrose, the isomerization of L-tagatose to L-galactose and D-ribulose to D-arabinose, and the isomerization of 6-deoxy-L-galactose to 6-deoxy-L-tagatose.

[0173] In some embodiments of the invention, L-rhamnose isomerase (EC 5.3.1.14) refers to an enzyme that catalyzes the reversible isomerization of 3R / 4S ketose to 2R / 3R / 4S aldose and the reversible isomerization of 3R / 4R ketose to 2R / 3R / 4R aldose. Isomerization reactions catalyzed by L-rhamnose isomerase include, but are not limited to, the isomerization of D-sorbose to D-gulose, the isomerization of D-psicose to D-allose, the isomerization of L-fructose to L-mannose and L-tagatose to L-talose, the isomerization of L-rhamnose (6-deoxy-L-mannose) to L-rhamnulose (6-deoxy-L-fructose), and the isomerization of 6-deoxy-L-psicose to 6-deoxy-L-allose.

[0174] In some embodiments of the present invention, D-lyxose isomerase (EC 5.3.1.15) refers to an enzyme that catalyzes the reversible isomerization of 3S / 4R ketose to 2S / 3S / 4R aldose and the reversible isomerization of 3S / 4S ketose to 2S / 3S / 4S aldose. Isomerization reactions catalyzed by D-lysine isomerase include, but are not limited to, the isomerization of D-mannose to D-fructose, the isomerization of D-xylulose to D-lyxose, the isomerization of L-psicose to L-allose, the isomerization of L-sorbose to L-gulose, the isomerization of L-ribulose to L-ribose, and the isomerization of L-tagatose to L-talose.

[0175] In some embodiments of the present invention, ribose isomerase (EC5.3.1.20) refers to an enzyme that catalyzes the reversible isomerization of 3R / 4R ketose to 2R / 3R / 4R aldose. Isomerization reactions catalyzed by ribose isomerase include but are not limited to the isomerization of D-ribose to D-ribulose, the isomerization of L-tagatose to L-talose, and the isomerization of D-psicose to D-allose.

[0176] In some embodiments of the present invention, D-psicose 3-epimerase (EC 5.1.3.30) refers to an enzyme that catalyzes the epimerization reaction of D-fructose at the C-3 position. The epimerization reaction catalyzed by D-psicose 3-epimerase includes, but is not limited to, the epimerization of D-fructose to D-psicose. In the present invention, the above-mentioned "enzyme that isomerizes D-fructose to D-psicose", for example, D-tagatose 3-epimerase of a certain microorganism that acts on D-fructose and converts it to D-psicose, is also included. The case where proteins produced by organisms such as all microorganisms, plants, and animals convert D-fructose to D-psicose is also included. The epimerization of other sugars and / or their derivatives also has the same meaning.

[0177] In some embodiments of the present invention, D-tagatose 3-epimerase (EC 5.1.3.31) refers to an enzyme that catalyzes the epimerization of various ketoses at the C3 position. The epimerization reactions catalyzed by D-tagatose 3-epimerase include but are not limited to the epimerization of D-fructose to D-psicose, the epimerization of D-tagatose to D-sorbose, the epimerization of D-ribulose to D-xylulose, and the epimerization of D-ribulose to D-xylulose. ose), L-ribulose is epimerized to L-xylulose, 6-deoxy-L-fructose is epimerized to 6-deoxy-L-psicose, 6-deoxy-L-tagatose is epimerized to 6-deoxy-L-sorbose, 1-deoxy-L-fructose is epimerized to 1-deoxy-L-psicose, and 1-deoxy-L-tagatose is epimerized to 1-deoxy-L-sorbose.

[0178] In some embodiments of the present invention, D-tagatose 4-epimerase can catalyze the epimerization of D-fructose to D-tagatose.

[0179] In some embodiments of the present invention, D-xylulose 4-epimerase can catalyze the epimerization of D-xylulose to L-ribulose.

[0180] In some embodiments of the present invention, ribose-5-phosphate isomerase (EC 5.3.1.6) refers to an enzyme that catalyzes the reversible isomerization of 3R / 4R phosphorylated ketose to 2R / 3R / 4R phosphorylated aldose. Isomerization reactions catalyzed by ribose-5-phosphate isomerase include, but are not limited to, catalyzing the isomerization of D-ribulose-5-phosphate to D-ribose-5-phosphate, the isomerization of D-ribulose to D-ribose, the isomerization of L-tagatose to L-talose, and the isomerization of D-psicose to D-allose. Ribose-5-phosphate isomerase generally shows specificity for D-ribose phosphate; however, due to structural similarities, the enzyme also shows broad substrate specificity for various other phosphorylated monosaccharides.

[0181] In some embodiments of the present invention, D-galactose 6-phosphate isomerase (EC 5.3.1.26) refers to an enzyme that catalyzes the reversible isomerization of 3S / 4S phosphorylated ketose to 2R / 3S / 4S phosphorylated aldose. Isomerization reactions catalyzed by D-galactose 6-phosphate include, but are not limited to, the isomerization of D-tagatose 6-phosphate to D-galactose 6-phosphate, the isomerization of D-tagatose to D-galactose, the isomerization of L-psicose to L-allose, and the isomerization of L-ribulose to L-arabinose. D-galactose 6-phosphate generally exhibits very close specificity for D-galactose phosphate; however, due to structural similarities, the enzyme also exhibits broad substrate specificity for a variety of other phosphorylated monosaccharides.

[0182] In some embodiments of the invention, D-glucose 6-phosphate isomerase (EC 5.3.1.9) is an enzyme that catalyzes the isomerization of D-glucose 6-phosphate to D-fructose 6-phosphate.

[0183] In some embodiments of the invention, mannose 6-phosphate isomerase (EC 5.3.1.8) is an enzyme that catalyzes the isomerization of D-mannose 6-phosphate to D-fructose 6-phosphate (F6P).

[0184] In some embodiments of the present invention, the D-psicose-6-phosphate 3-epimerase is an enzyme that catalyzes the epimerization of D-fructose 6-phosphate to D-psicose 6-phosphate.

[0185] In some embodiments of the present invention, the D-tagatose 6-phosphate 4-epimerase is an enzyme that catalyzes the epimerization of D-fructose 6-phosphate to D-tagatose 6-phosphate.

[0186] In some embodiments of the invention, L-ribulose-5-phosphate-4-epimerase (EC 5.1.3.4) is an enzyme that catalyzes the isomerization of L-ribulose 5-phosphate to D-xylulose 5-phosphate.

[0187] In some embodiments of the present invention, cellobiose-2-epimerase (EC 5.1.3.11) refers to an enzyme that catalyzes the interconversion between the reducing end glucosyl and mannosyl of a disaccharide linked by a β-1,4 glycosidic bond. The epimerization reaction catalyzed by cellobiose-2-epimerase includes, but is not limited to, catalyzing the epimerization of cellobiose to 4-O-β-D-pyranoglucopyranosyl-D-mannose, the isomerization of cellobiose to 4-O-β-D-pyranoglucopyranosyl-D-fructose, the epimerization of D-glucose to D-mannose, the isomerization of D-glucose to D-fructose, the isomerization of lactose to lactulose, and the epimerization of lactose to epilactose.

[0188] In some embodiments of the invention, amylosucrase (EC 2.4.1.4) can catalyze the isomerization of sucrose to turanose.

[0189] In some embodiments of the invention, sucrose glucosylmutase (EC 5.4.99.11) can catalyze the isomerization of sucrose to isomaltulose and trehalulose.

[0190] In some embodiments of the present invention, trehalose synthase can catalyze the isomerization of sucrose into trehalulose, and can also catalyze the isomerization of maltobiose into trehalose.

[0191] The component (c) enzyme of the present invention can be used in various forms, including but not limited to purified or partially purified enzymes, microorganisms and / or microbial cells expressing the enzymes, cultures of the microorganisms, protein extracts of the microorganisms, protein extracts of cultures of the microorganisms, or combinations thereof.

[0192] In some embodiments, the component (c) enzyme used in the boron-containing composition can be selected according to the substrate used and the target product. In some embodiments, the enzyme comprises one, two, three or more enzymes having the same or different functions.

[0193] In some exemplary embodiments, when the boron-containing composition is used to catalyze the preparation of monosaccharides, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one monosaccharide into other monosaccharides.

[0194] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-fructose from D-glucose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes D-glucose to D-fructose.

[0195] In the present invention, the isomerase that isomerizes D-glucose to D-fructose includes but is not limited to xylose isomerase (EC5.3.1.5).

[0196] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-xylulose from D-xylose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-xylose to D-xylulose.

[0197] In the present invention, the isomerase that isomerizes D-xylose to D-xylulose includes but is not limited to xylose isomerase (EC5.3.1.5).

[0198] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-psicose from D-fructose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an epimerase that isomerizes D-fructose to D-psicose.

[0199] In the present invention, the epimerase that isomerizes D-fructose to D-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31) and D-psicose 3-epimerase (EC 5.1.3.30).

[0200] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-tagatose from D-galactose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes D-galactose to D-tagatose.

[0201] In the present invention, the isomerase that isomerizes D-galactose to D-tagatose includes but is not limited to L-arabinose isomerase (EC 5.3.1.4) and D-galactose 6-phosphate isomerase (EC 5.3.1.26).

[0202] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-psicose from D-glucose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose to D-fructose and an epimerase that isomerizes D-fructose to D-psicose.

[0203] In the present invention, the isomerase that isomerizes D-glucose into D-fructose includes but is not limited to xylose isomerase (EC5.3.1.5); the epimerase that isomerizes D-fructose into D-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31) and D-psicose 3-epimerase (EC 5.1.3.30).

[0204] In some exemplary embodiments, when the boron-containing composition catalyzes the production of L-ribulose from D-xylose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-xylose to D-xylulose and an epimerase that isomerizes D-xylulose to L-ribulose.

[0205] In the present invention, the isomerase that isomerizes D-xylose to D-xylulose includes but is not limited to xylose isomerase (EC5.3.1.5); the epimerase that isomerizes D-xylulose to L-ribulose includes but is not limited to D-xylulose 4-epimerase.

[0206] In some exemplary embodiments, when the boron-containing composition catalyzes the preparation of phosphate monosaccharides, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes phosphate monosaccharides into other phosphate monosaccharides.

[0207] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-glucose 6-phosphate from D-fructose 6-phosphate, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate.

[0208] In the present invention, the isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate includes but is not limited to D-glucose 6-phosphate isomerase (EC 5.3.1.9).

[0209] In the present invention, the enzyme of component (c) in the boron-containing composition or the boron-containing composition catalyst may further include phosphoglucomutase to convert glucose 1-phosphate into glucose 6-phosphate.

[0210] In the present invention, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst may also include an enzyme that converts sugar into glucose 1-phosphate, such as α-glucan phosphorylase, maltose phosphorylase, sucrose phosphorylase, cellodextrin phosphorylase, cellobiose phosphorylase and / or cellulose phosphorylase, and mixtures thereof.

[0211] In some exemplary embodiments, when the boron-containing composition catalyzes the production of D-fructose 6-phosphate from sugar (starch and / or its derivatives, cellulose and / or its derivatives, fructose, glucose and / or sucrose), the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an enzyme that converts sugar into glucose 1-phosphate, a phosphoglucomutase, and an isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate.

[0212] In some exemplary embodiments, when the boron-containing composition catalyzes the preparation of D-psicose 6-phosphate from D-glucose 6-phosphate, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate and an isomerase that epimerizes D-fructose 6-phosphate to D-psicose 6-phosphate.

[0213] In the present invention, the isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate includes but is not limited to D-glucose 6-phosphate isomerase (EC 5.3.1.9); the isomerase that epimerizes D-fructose 6-phosphate to D-psicose 6-phosphate includes but is not limited to D-psicose-6-phosphate 3-position epimerase.

[0214] In some exemplary embodiments, when the boron-containing composition catalyzes the production of deoxy sugars, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one deoxy sugar to other deoxy sugars.

[0215] In some exemplary embodiments, when the boron-containing composition catalyzes the preparation of L-rhamnose (6-deoxyL-mannose) from L-rhamnulose (6-deoxyL-fructose), the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes L-rhamnose (6-deoxyL-mannose) to L-rhamnulose (6-deoxyL-fructose).

[0216] In the present invention, the isomerase that isomerizes L-rhamnose (6-deoxy L-mannose) to L-rhamnulose (6-deoxy L-fructose) includes but is not limited to L-rhamnose isomerase (EC5.3.1.14).

[0217] In some exemplary embodiments, when the boron-containing composition catalyzes the production of 6-deoxyL-tagatose from 6-deoxyL-galactose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes 6-deoxyL-galactose to 6-deoxyL-tagatose.

[0218] In the present invention, the isomerase that isomerizes 6-deoxyL-galactose to 6-deoxyL-tagatose includes but is not limited to D-arabinose isomerase (EC5.3.1.3).

[0219] In some exemplary embodiments, when the boron-containing composition catalyzes the production of 6-deoxyL-psicose from 6-deoxyL-fructose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes 6-deoxyL-fructose to 6-deoxyL-psicose.

[0220] In the present invention, the isomerase that epimerizes 6-deoxyL-fructose to 6-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0221] In some exemplary embodiments, when the boron-containing composition catalyzes 1-deoxyL-fructose to produce 1-deoxyL-psicose, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes 1-deoxyL-fructose to 1-deoxyL-psicose.

[0222] In the present invention, the isomerase that epimerizes 1-deoxyL-fructose to 1-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0223] In some exemplary embodiments, when the boron-containing composition catalyzes the preparation of 6-deoxyL-psicose from L-rhamnose (6-deoxyL-mannose), the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes L-rhamnose (6-deoxyL-mannose) to L-rhamnulose (6-deoxyL-fructose) and an isomerase that isomerizes 6-deoxyL-fructose to 6-deoxyL-psicose.

[0224] In the present invention, the isomerase that isomerizes L-rhamnose (6-deoxyL-mannose) to L-rhamnulose (6-deoxyL-fructose) includes but is not limited to L-rhamnose isomerase (EC5.3.1.14); the isomerase that epimerizes 6-deoxyL-fructose to 6-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0225] In some exemplary embodiments, when the boron-containing composition catalyzes the preparation of disaccharides, the component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes disaccharides into other disaccharides.

[0226] In some exemplary embodiments, when the boron-containing composition catalyzes lactose to produce lactulose, the enzyme of component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes lactose into lactulose.

[0227] In the present invention, the isomerase that isomerizes lactose into lactulose includes but is not limited to cellobiose-2-epimerase (EC 5.1.3.11).

[0228] In some exemplary embodiments, when the boron-containing composition catalyzes the production of turanoose from sucrose, the enzyme of component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes sucrose to turanoose.

[0229] In the present invention, the isomerase that isomerizes sucrose to turanoose includes but is not limited to amylosucrase (EC 2.4.1.4).

[0230] Other components

[0231] In the present invention, the boron-containing composition may further include a connector. The connector carries an anchoring group to attach the component (b) boric acid compound and / or its derivative and / or the component (c) enzyme to the component (a) matrix to achieve the loading of the component (b) boric acid compound and / or its derivative and / or the component (c) enzyme on the component (a) matrix. In some embodiments of the present invention, the connector may be a cross-linking agent. Examples of the cross-linking agent can be selected from glutaraldehyde, epichlorohydrin, polyfunctional aziridine, N,N-methylenebisacrylamide, bifunctional carbodiimide (e.g., dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, N-hydroxysulfosuccinimide), ethylene glycol bis(succinimidyl succinate), ethylene glycol bis(sulfosuccinimidyl succinate), tris-(succinimidyl)aminotriacetate, dimethyl pipecolate, dimethyl suberoyl, dimethyl 3,3'-dithiobispropionimide, trishydroxymethylphosphine, NHS-phosphine, NHS-PEG-azide, NHS-azide, genipin, dopamine, polydopamine or a combination thereof.

[0232] Load mode

[0233] In some embodiments of the present invention, the component (b) boronic acid compound and / or its derivative and the component (c) enzyme are respectively loaded on the same component (a) matrix, such as Figure 1 In some embodiments of the present invention, the component (b) boronic acid compound and / or its derivative and the component (c) enzyme are respectively loaded on different component (a) matrices, such as Figure 1In some embodiments of the present invention, the component (b) boronic acid compound and / or its derivative and the component (c) enzyme are simultaneously loaded on the same component (a) matrix, such as Figure 1 In some embodiments of the present invention, the component (b) boronic acid compound is loaded on the component (a) matrix, and the component (c) enzyme can be in any free form, such as Figure 1 As shown in D in FIG. In the present invention, the loading can be achieved by adsorption, embedding, cross-linking, covalent bonding or a combination thereof.

[0234] In the present invention, in order to prepare different carbohydrate compounds from inexpensive substrates by isomerization, optionally, one functional enzyme is loaded on the component (a) matrix, two functional enzymes are loaded on the component (a) matrix, three functional enzymes are loaded on the component (a) matrix, or more functional enzymes are loaded on the component (a) matrix.

[0235] <Method for preparing boron-containing composition>

[0236] The second aspect of the present invention provides a method for preparing the boron-containing composition of the first aspect of the present invention, the method comprising the steps of:

[0237] The component (b) boric acid compound and / or its derivative and / or the component (c) enzyme containing free species are mixed with the substrate (a) and loaded to obtain the boron-containing composition of the first aspect of the present invention.

[0238] In some embodiments of the present invention, the preparation method of the boron-containing composition further comprises the step of adding a linker. In some embodiments of the present invention, the linker can be a cross-linking agent. The example of the cross-linking agent can be selected from glutaraldehyde, epichlorohydrin, polyfunctional aziridine, N, N-methylenebisacrylamide, bifunctional carbodiimide (e.g., dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, N-hydroxysuccinimide, N-hydroxysulfosuccinimide), ethylene glycol bis (succinimidyl succinate), ethylene glycol bis (sulfosuccinimidyl succinate), tris-(succinimidyl) aminotriacetate, dimethyl pipecolate, dimethyl suberimidate, dimethyl 3,3'-dithiobispropionimidate, trimethylol phosphorus, NHS-phosphine, NHS-PEG-azide, NHS-azide, genipin, dopamine, polydopamine or a combination thereof.

[0239] <Use of boron-containing composition in the preparation of sugar and / or its derivatives / as a catalyst for the preparation of sugar and / or its derivatives>

[0240] The third aspect of the present invention provides use of the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention in preparing sugar and / or its derivatives.

[0241] The third aspect of the present invention further provides use of the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention as a catalyst for preparing sugars and / or their derivatives.

[0242] The boron-containing composition of the present invention can be used under acidic conditions, neutral conditions or alkaline conditions (for example, for preparing sugars and / or their derivatives). In some specific embodiments of the present invention, the acidic condition can be a range of pH values ​​below 7.0, the neutral condition is a pH value equal to 7.0, and the alkaline condition is a range of pH values ​​above 7.0; in some specific embodiments, the pH value is between 7.0 and 9.0; in some more specific embodiments, the pH value is between 7.0 and 8.5; illustratively, the pH value can be 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4 or 8.5.

[0243] The boron-containing composition of the present invention can be used in a solution (for example, for preparing sugar and / or its derivatives). In some specific embodiments, the boron-containing composition of the present invention can be used in any solution that can provide the above-mentioned suitable pH (for example, for preparing sugar and / or its derivatives), for example, it can be selected from aqueous solutions, organic solvents, ionic liquids, deep eutectic solvents, etc., and for example, the solution can be a buffer solution or a non-buffer solution. Exemplarily, the buffer can be an acetate buffer (e.g., acetic acid-sodium acetate buffer), a citric acid buffer (e.g., citric acid-sodium citrate buffer), a glycine buffer (e.g., glycine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, etc.), a phthalate buffer (e.g., potassium hydrogen phthalate-sodium hydroxide buffer), a phosphate buffer (e.g., disodium hydrogen phosphate-sodium dihydrogen phosphate buffer, dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer, potassium dihydrogen phosphate-sodium hydroxide buffer, etc.), 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) buffer, 3-morpholinepropanesulfonic acid (MOPS) buffer, tris(hydroxymethylaminomethane) (Tris) buffer, a carbonate buffer (e.g., sodium carbonate-sodium bicarbonate buffer), an ammonia-ammonium chloride buffer, and the like.

[0244] In some embodiments of the present invention, the sugar comprises one or more of a monosaccharide, a disaccharide, an oligosaccharide, and a polysaccharide.

[0245] In some preferred embodiments of the present invention, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides.

[0246] In some preferred embodiments of the present invention, the sugar includes one or more of monosaccharides and disaccharides.

[0247] In some embodiments of the present invention, the sugar derivatives include one or more of phosphate sugars and deoxy sugars.

[0248] In some embodiments of the present invention, the boron-containing composition is used to catalyze the preparation of monosaccharides. For example, the boron-containing composition can be used to catalyze glucose to produce fructose, to catalyze fructose to produce psicose, to catalyze xylose to produce xylulose, to catalyze D-xylulose to produce L-ribulose, and to catalyze galactose to produce tagatose. For example, the boron-containing composition can also be used to catalyze glucose to produce psicose via fructose. For example, the boron-containing composition can also be used to catalyze D-xylose to produce L-ribulose via D-xylulose.

[0249] In some embodiments of the present invention, the boron-containing composition is used to prepare phosphate monosaccharides. For example, the boron-containing composition can be used to catalyze glucose 6-phosphate to produce fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce psicose 6-phosphate via fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce tagatose 6-phosphate via fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce mannose 6-phosphate via fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce galactose 6-phosphate via fructose 6-phosphate and tagatose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce talose 6-phosphate via fructose 6-phosphate and tagatose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce sorbose 6-phosphate via fructose 6-phosphate and tagatose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce idose 6-phosphate via fructose 6-phosphate, tagatose 6-phosphate and sorbose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce gulose 6-phosphate via fructose 6-phosphate, tagatose 6-phosphate and sorbose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce allose 6-phosphate via fructose 6-phosphate and psicose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce altrose 6-phosphate via fructose 6-phosphate and psicose 6-phosphate.

[0250] In some embodiments of the present invention, the boron-containing composition is used to catalyze glucose 6-phosphate to prepare monosaccharides and derivatives via phosphate monosaccharides. For example, the boron-containing composition can be used to catalyze glucose 6-phosphate to prepare fructose via fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to prepare psicose via fructose 6-phosphate and psicose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to prepare tagatose via fructose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to prepare mannose via fructose 6-phosphate and mannose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to prepare galactose via fructose 6-phosphate, tagatose 6-phosphate and galactose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to prepare talose via fructose 6-phosphate, tagatose 6-phosphate and talose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce sorbose via fructose 6-phosphate, tagatose 6-phosphate and sorbose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce idose via fructose 6-phosphate, tagatose 6-phosphate, sorbose 6-phosphate and idose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce gulose via fructose 6-phosphate, tagatose 6-phosphate, sorbose 6-phosphate and gulose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce allose via fructose 6-phosphate, psicose 6-phosphate and allose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce altrose via fructose 6-phosphate, psicose 6-phosphate and altrose 6-phosphate. For example, the boron-containing composition can also be used to catalyze glucose 6-phosphate to produce fructose 1,6-diphosphate via fructose 6-phosphate.

[0251] In the present invention, the glucose 6-phosphate can be prepared from glucose 1-phosphate.

[0252] In the present invention, the glucose 1-phosphate can be prepared from saccharides (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) through one or more steps of reaction, wherein the one or more steps of reaction are catalyzed by at least one enzyme, a microorganism expressing the enzyme and / or a culture of a microorganism expressing the enzyme, and the saccharide can be selected from, but not limited to, starch and / or its derivatives, cellulose and / or its derivatives, fructose, glucose and / or sucrose. According to the method of the present disclosure, the one or more enzymes used in the step of converting saccharides into glucose 1-phosphate can be α-glucan phosphorylase, maltose phosphorylase, sucrose phosphorylase, cellodextrin phosphorylase, cellobiose phosphorylase and / or cellulose phosphorylase, and mixtures thereof, and / or microorganisms expressing the above one or more enzymes and mixtures thereof, and / or cultures of microorganisms expressing the above one or more enzymes and mixtures thereof.

[0253] When the carbohydrate is starch and / or its derivatives, the starch and / or its derivatives can be selected from, including but not limited to, amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, maltose, glucose and mixtures thereof. In certain embodiments of the present disclosure, when the carbohydrate is starch and / or its derivatives, including but not limited to amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, the enzyme for converting the carbohydrate into glucose 1-phosphate comprises α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase.

[0254] In the present invention, when the glucose 1-phosphate is prepared from carbohydrates (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) through one or more steps of reaction, the starch derivative can be prepared by enzymatic hydrolysis of starch and / or acid hydrolysis of starch. In certain embodiments of the present disclosure, in order to increase the yield of the product, starch can be converted to dextrin using starch branching enzymes such as isoamylase (IA), microorganisms expressing isoamylase and / or cultures of microorganisms expressing isoamylase. In certain embodiments of the present disclosure, in order to increase the yield of the product, starch can also be converted to dextrin using starch branching enzymes such as pullulanase (PA), microorganisms expressing pullulanase and / or cultures of microorganisms expressing pullulanase.

[0255] In some other embodiments of the present disclosure, in order to increase the yield of the product, the degradation products of starch and / or its derivatives, glucose, maltose and maltotriose, can be converted into longer maltooligosaccharides using 4-glucanotransferase (4GT), microorganisms expressing 4-glucanotransferase and / or cultures of microorganisms expressing 4-glucanotransferase, wherein the longer maltooligosaccharides can be converted into glucose 1-phosphate using α-glucan phosphorylase, microorganisms expressing α-glucan phosphorylase and / or cultures of microorganisms expressing α-glucan phosphorylase. In some further embodiments of the present disclosure, in order to increase the yield of the product, the degradation product of starch and / or its derivatives, glucose, can be converted into glucose 6-phosphate by adding polyphosphate using polyphosphate glucokinase (PPGK), microorganisms expressing polyphosphate glucokinase and / or cultures of microorganisms expressing polyphosphate glucokinase.

[0256] When the carbohydrate is maltose and / or maltooligosaccharide, the enzyme for converting the carbohydrate into β-glucose 1-phosphate comprises maltose phosphorylase, a microorganism expressing maltose phosphorylase, and / or a culture of a microorganism expressing maltose phosphorylase. The β-glucose 1-phosphate is further converted into glucose 6-phosphate by β-phosphoglucose mutase (β-PGM), a microorganism expressing β-phosphoglucose mutase, and / or a culture of a microorganism expressing β-phosphoglucose mutase.

[0257] When the carbohydrate is sucrose, the enzyme for converting the carbohydrate into glucose 1-phosphate comprises sucrose phosphorylase, a microorganism expressing sucrose phosphorylase and / or a culture of a microorganism expressing sucrose phosphorylase. In some other embodiments of the present disclosure, in order to increase the output of the product, the degradation product fructose of sucrose can be converted into glucose using glucose isomerase (GI), a microorganism expressing glucose isomerase and / or a culture of a microorganism expressing glucose isomerase. Glucose and polyphosphate are then catalyzed to generate glucose 1-phosphate by polyphosphate glucokinase.

[0258] When the sugar is cellulose and / or its derivatives, the cellulose and / or its derivatives can be selected from non-edible lignocellulosic materials (e.g., cellulose, hemicellulose and / or lignin and other minor components), pure cellulose (Avicel (microcrystalline cellulose), regenerated amorphous cellulose, bacterial cellulose, filter paper, etc.), partially hydrolyzed cellulose substrates (including water-insoluble cellodextrins with a degree of polymerization greater than 7, water-soluble cellodextrins with a degree of polymerization of 3-6, cellobiose, glucose and fructose). In certain embodiments of the present disclosure, when the sugar is cellulose and its derivatives, the enzymes used to convert the sugar into glucose 1-phosphate include cellodextrin phosphorylase (CDP), a microorganism expressing cellodextrin phosphorylase and / or a culture of a microorganism expressing cellodextrin phosphorylase, and cellobiose phosphorylase (CBP), a microorganism expressing cellobiose phosphorylase and / or a culture of a microorganism expressing cellobiose phosphorylase.

[0259] In the present invention, when the glucose 1-phosphate is prepared from carbohydrates (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) through one or more steps of reaction, the cellulose derivative can be prepared by cellulose, for example, solid cellulose is hydrolyzed into water-soluble cellodextrins and cellobiose by using endoglucanase, microorganisms expressing endoglucanase and / or cultures of microorganisms expressing endoglucanase and / or cellobiohydrolases, microorganisms expressing cellobiohydrolases and / or cultures of microorganisms expressing cellobiohydrolases. In some embodiments, cellulose can be pretreated before cellulose is hydrolyzed to generate glucose 1-phosphate to increase their reactivity and reduce the degree of polymerization of cellulose chains. The pretreatment method of the cellulose includes but is not limited to dilute acid pretreatment, lignocellulose fractionation based on cellulose solvent, ammonia fiber expansion, ammonia soaking, ionic liquid treatment, and partial hydrolysis by using concentrated acid (including hydrochloric acid, sulfuric acid, phosphoric acid and combinations thereof).

[0260] In certain embodiments of the present disclosure, in order to increase the yield of the product, cellobiose, a degradation product of cellulose and / or its derivatives, can be converted into glucose 1-phosphate and glucose using cellobiose phosphorylase, a microorganism expressing cellobiose phosphorylase, and / or a culture of a microorganism expressing cellobiose phosphorylase. In some further embodiments of the present disclosure, in order to increase the yield of the product, polyphosphate glucokinase (PPGK), a microorganism expressing polyphosphate glucokinase, and / or a culture of a microorganism expressing polyphosphate glucokinase can be used to convert glucose, a degradation product of cellulose and / or its derivatives, into glucose 6-phosphate by adding polyphosphate.

[0261] In some embodiments of the present invention, the boron-containing composition is used to catalyze the preparation of deoxy sugars, particularly 6-deoxy sugars. For example, the boron-containing composition can catalyze the isomerization of L-rhamnose (6-deoxy L-mannose) to L-rhamnulose (6-deoxy L-fructose). For example, the boron-containing composition can catalyze the epimerization of 6-deoxy L-fructose to 6-deoxy L-psicose. For example, the boron-containing composition can catalyze the epimerization of 1-deoxy L-fructose to 1-deoxy L-psicose. For example, the boron-containing composition can catalyze the isomerization of 6-deoxy L-galactose to 6-deoxy L-tagatose.

[0262] In some embodiments of the present invention, the boron-containing composition is used to prepare disaccharides. For example, the boron-containing composition can be used to catalyze lactose to produce lactulose. For example, the boron-containing composition can be used to catalyze sucrose to produce trehalulose. For example, the boron-containing composition can be used to catalyze sucrose to produce isomaltulose. For example, the boron-containing composition can be used to catalyze sucrose to produce turanoose.

[0263] Catalytic process using boron-containing compositions

[0264] The fourth aspect of the present invention provides a method for a reaction catalyzed by the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention, the method comprising the steps of:

[0265] (i) preparing a reaction mixture comprising a substrate,

[0266] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0267] In some embodiments of the present invention, a method for preparing sugar and / or its derivatives is provided, wherein the method uses the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention as a catalyst, and the method comprises the steps of:

[0268] (i) preparing a reaction mixture comprising a substrate,

[0269] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0270] In the present invention, the contacting may be carried out by passing the reaction mixture through a packed bed column of the boron composition or boron-containing composition catalyst; may be carried out in a moving bed reactor in which a packed bed of the boron composition or boron-containing composition catalyst is moved co-currently or counter-currently with the reaction mixture; may be carried out in a batch reactor, a continuous stirred tank reactor in which the boron composition or boron-containing composition catalyst is kept in solid form, optionally under stirring, or in any other type of reactor or reactors or combinations thereof in which the desired reaction can be carried out.

[0271] In some embodiments of the present invention, when a reaction catalyzed by the boron-containing composition or boron-containing composition catalyst of the present invention is carried out in a (stirred) tank reactor, the boron-containing composition or boron-containing composition catalyst can be subsequently separated or recovered from the reactants by filtration, centrifugation, etc. After separation, the boron-containing composition or boron-containing composition catalyst can be used again (recycled) in the process.

[0272] In some embodiments of the present invention, the reaction mixture may be contacted with other catalysts or enzymes while contacting the reaction mixture with the boron-containing composition or boron-containing composition catalyst under suitable reaction conditions. In other embodiments of the present invention, the product may be contacted with other catalysts or enzymes after contacting the reaction mixture with the boron-containing composition or boron-containing composition catalyst under suitable reaction conditions.

[0273] In the method for the reaction catalyzed by the boron-containing composition of the present invention, the reaction mixture is contacted with the boron-containing composition or the boron-containing composition catalyst at a certain temperature. The suitable temperature depends on factors such as the nature and amount of the boron-containing composition and the nature and amount of the substrate.

[0274] In some specific embodiments, the temperature is between 20°C and 95°C. In some more specific embodiments, the temperature is between 25°C and 95°C, 30°C and 95°C, 35°C and 95°C, 35°C and 90°C, 35°C and 85°C, 35°C and 80°C, or 40°C and 80°C. Exemplarily, the temperature is 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, or 80°C.

[0275] In the method of the boron-containing composition catalyzed reaction of the present invention, the reaction mixture is contacted with the boron-containing composition or the boron-containing composition catalyst at a certain pH. The suitable pH depends on factors such as the nature and amount of the boron-containing composition and the nature and amount of the substrate.

[0276] In the present invention, the suitable pH value may be an acidic condition, a neutral condition or an alkaline condition. In some specific embodiments of the present invention, the acidic condition may be a pH value in the range below 7.0, the neutral condition is a pH value equal to 7.0, and the alkaline condition is a pH value in the range above 7.0; in some specific embodiments, the pH value is between 7.0 and 9.0; in some more specific embodiments, the pH value is between 7.0 and 8.5; illustratively, the pH value may be 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4 or 8.5.

[0277] In the present invention, the suitable reaction conditions include a suitable solution, and the suitable solution depends on factors such as the nature and amount of the boron-containing composition and the nature and amount of the substrate. In the present invention, there is no particular limitation on the type of solution, for example, it can be any solution that can provide the above-mentioned suitable pH, for example, it can be selected from aqueous solutions, organic solvents, ionic liquids, deep eutectic solvents, etc., and for example, the solution can be a buffer solution or a non-buffer solution. Exemplarily, the buffer can be an acetate buffer (e.g., acetic acid-sodium acetate buffer), a citric acid buffer (e.g., citric acid-sodium citrate buffer), a glycine buffer (e.g., glycine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, etc.), a phthalate buffer (e.g., potassium hydrogen phthalate-sodium hydroxide buffer), a phosphate buffer (e.g., disodium hydrogen phosphate-sodium dihydrogen phosphate buffer, dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer, potassium dihydrogen phosphate-sodium hydroxide buffer, etc.), 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) buffer, 3-morpholinepropanesulfonic acid (MOPS) buffer, tris(hydroxymethylaminomethane) (Tris) buffer, a carbonate buffer (e.g., sodium carbonate-sodium bicarbonate buffer), an ammonia-ammonium chloride buffer, and the like.

[0278] In the method for the reaction catalyzed by the boron-containing composition of the present invention, the solution contains metal ions. For example, the metal ions may be magnesium ions, cobalt ions, and the like.

[0279] In the method for reaction catalyzed by the boron-containing composition of the present invention, in the reaction system (or preparation system, catalytic system), the amount of the boron-containing composition or the boron-containing composition catalyst is 0.1 g / L to 30 g / L, preferably 0.1 g / L to 25 g / L, 0.1 g / L to 20 g / L, 0.3 g / L to 20 g / L, 0.5 g / L to 20 g / L. Exemplarily, in the reaction system (or preparation system, catalytic system), the amount of the boron-containing composition or the boron-containing composition catalyst is 0.5 g / L, 1.0 g / L, 1.5 g / L, 2.0 g / L, 2.5 g / L, 3.0 g / L, 3.5 g / L, 4 g / L, 4.5 g / L, 5.0 g / L, 6.0 g / L, 7.0 g / L, 8.0 g / L, 9 g / L, 10 g / L, 12 g / L, 14 g / L, 16 g / L, 18 g / L, or 20 g / L.

[0280] In the method for the reaction catalyzed by the boron-containing composition of the present invention, in the reaction system (or preparation system, catalytic system), the amount of the substrate is 10-5000mM, preferably 10-4000mM, 10-3000mM, 10-2000mM, 20-2000mM, 30-2000mM, 40-2000mM, 50-2000mM. Exemplarily, in the reaction system (or preparation system, catalytic system), the amount of the substrate is 50mM, 55mM, 100mM, 200mM, 278mM, 300mM, 333mM, 500mM, 700mM, 900mM, 1000mM, 2000mM.

[0281] In the method of the boron-containing composition catalyzed reaction of the present invention, the reaction mixture is contacted with the boron-containing composition or the boron-containing composition catalyst for a period of time. The appropriate reaction time depends on factors such as the nature and amount of the boron-containing composition and the nature and amount of the substrate.

[0282] Preferably, the combination of temperature, pH and / or reaction time is effective to ensure a higher product to substrate ratio.

[0283] In some embodiments of the invention, the substrate comprises a sugar and / or a derivative thereof.

[0284] In some alternative embodiments, the sugar as the substrate includes one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides.

[0285] In some alternative embodiments, the derivative of the sugar as the substrate includes one or more of phosphate sugar and deoxy sugar.

[0286] Exemplarily, the substrate is a monosaccharide (e.g., glucose, xylose, fructose, galactose), a disaccharide (e.g., lactose, sucrose), a phosphate monosaccharide (e.g., glucose 6-phosphate), a 6-deoxy sugar (e.g., L-rhamnose / 6-deoxyL-mannose), or a 1-deoxy sugar (e.g., 1-deoxyL-fructose).

[0287] In some embodiments of the present invention, a method for catalytically preparing monosaccharides using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0288] (i) preparing a reaction mixture comprising a substrate;

[0289] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions;

[0290] The component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one monosaccharide into other monosaccharides.

[0291] In some specific examples of the present invention, a method for preparing D-fructose by catalyzing D-glucose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0292] (i) preparing a reaction mixture comprising D-glucose;

[0293] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0294] The enzyme as component (c) in the boron-containing composition or boron-containing composition catalyst comprises an isomerase that isomerizes D-glucose into D-fructose.

[0295] In the present invention, the isomerase that isomerizes D-glucose to D-fructose includes but is not limited to xylose isomerase (EC5.3.1.5).

[0296] In some specific examples of the present invention, a method for preparing D-xylulose by catalyzing D-xylose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0297] (i) preparing a reaction mixture comprising D-xylose;

[0298] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0299] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes D-xylose into D-xylulose.

[0300] In the present invention, the isomerase that isomerizes D-xylose to D-xylulose includes but is not limited to xylose isomerase (EC5.3.1.5).

[0301] In some specific examples of the present invention, a method for preparing D-psicose by catalyzing D-fructose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0302] (i) preparing a reaction mixture comprising D-fructose;

[0303] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0304] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst includes an epimerase that isomerizes D-fructose into D-psicose.

[0305] In the present invention, the epimerase that isomerizes D-fructose to D-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31) and D-psicose 3-epimerase (EC 5.1.3.30).

[0306] In some specific examples of the present invention, a method for preparing D-tagatose by catalyzing D-galactose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0307] (i) preparing a reaction mixture comprising D-galactose;

[0308] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0309] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes D-galactose into D-tagatose.

[0310] In the present invention, the isomerase that isomerizes D-galactose to D-tagatose includes but is not limited to L-arabinose isomerase (EC 5.3.1.4) and D-galactose 6-phosphate isomerase (EC 5.3.1.26).

[0311] In some specific examples of the present invention, a method for preparing D-psicose by catalyzing D-glucose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0312] (i) preparing a reaction mixture comprising D-glucose;

[0313] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0314] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose into D-fructose and an epimerase that isomerizes D-fructose into D-psicose.

[0315] In the present invention, the isomerase that isomerizes D-glucose into D-fructose includes but is not limited to xylose isomerase (EC5.3.1.5); the epimerase that isomerizes D-fructose into D-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31) and D-psicose 3-epimerase (EC 5.1.3.30).

[0316] In some specific examples of the present invention, a method for preparing L-ribulose from D-xylose by catalyzing the boron-containing composition prepared according to the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0317] (i) preparing a reaction mixture comprising D-xylose;

[0318] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0319] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-xylose into D-xylulose and an epimerase that isomerizes D-xylulose into L-ribulose.

[0320] In the present invention, the isomerase that isomerizes D-xylose to D-xylulose includes but is not limited to xylose isomerase (EC5.3.1.5); the epimerase that isomerizes D-xylulose to L-ribulose includes but is not limited to D-xylulose 4-epimerase.

[0321] In some embodiments of the present invention, a method for catalyzing the preparation of phosphate monosaccharides using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0322] (i) preparing a reaction mixture comprising a substrate;

[0323] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions;

[0324] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one phosphate monosaccharide into other phosphate monosaccharides.

[0325] In some specific examples of the present invention, a method for preparing D-fructose 6-phosphate by catalyzing D-glucose 6-phosphate using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0326] (i) preparing a reaction mixture comprising D-glucose 6-phosphate;

[0327] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0328] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose 6-phosphate into D-fructose 6-phosphate.

[0329] In the present invention, the isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate includes but is not limited to D-glucose 6-phosphate isomerase (EC 5.3.1.9).

[0330] In the present invention, the method for preparing D-fructose 6-phosphate by catalyzing D-glucose 6-phosphate with the boron-containing composition may also include the step of converting glucose 1-phosphate into glucose 6-phosphate by utilizing phosphoglucomutase, a microorganism expressing phosphoglucomutase and / or a culture of a microorganism expressing phosphoglucomutase.

[0331] In the present invention, the method for preparing D-fructose 6-phosphate by catalyzing D-glucose 6-phosphate with the boron-containing composition may further include the step of converting saccharides (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) into glucose 1-phosphate, wherein the step is catalyzed by at least one enzyme, a microorganism expressing the enzyme, and / or a culture of a microorganism expressing the enzyme, and the saccharide may be selected from, but not limited to, starch and / or its derivatives, cellulose and / or its derivatives, fructose, glucose and / or sucrose. According to the method disclosed herein, the one or more enzymes used in the step of converting saccharides into glucose 1-phosphate may be α-glucan phosphorylase, maltose phosphorylase, sucrose phosphorylase, cellodextrin phosphorylase, cellobiose phosphorylase and / or cellulose phosphorylase, and mixtures thereof, and / or microorganisms expressing the above one or more enzymes and mixtures thereof, and / or cultures of microorganisms expressing the above one or more enzymes and mixtures thereof.

[0332] When the carbohydrate is starch and / or its derivatives, the starch and / or its derivatives can be selected from, including but not limited to, amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, maltose, glucose and mixtures thereof. In certain embodiments of the present disclosure, when the carbohydrate is starch and / or its derivatives, including but not limited to amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, the enzyme for converting the carbohydrate into glucose 1-phosphate comprises α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase.

[0333] Some methods of the present disclosure may further include the step of converting starch into starch derivatives, wherein the starch derivatives are prepared by enzymatic hydrolysis of starch and / or acid hydrolysis of starch. In certain embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting starch into dextrins using a starch branching enzyme such as isoamylase (IA), a microorganism expressing isoamylase, and / or a culture of a microorganism expressing isoamylase. In certain embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the more excellent method for preparing fructose 6-phosphate may further include converting starch into dextrins using a starch branching enzyme such as pullulanase (PA), a microorganism expressing pullulanase, and / or a culture of a microorganism expressing pullulanase.

[0334] In some other embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with the more excellent boron-containing composition may further include using 4-glucan transferase (4GT), a microorganism expressing 4-glucan transferase and / or a culture of a microorganism expressing 4-glucan transferase to convert the degradation products of starch and / or its derivatives, glucose, maltose and maltotriose, into longer maltooligosaccharides, wherein the longer maltooligosaccharides can be converted into glucose 1-phosphate using α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include using polyphosphate glucose kinase (PPGK), a microorganism expressing polyphosphate glucose kinase and / or a culture of a microorganism expressing polyphosphate glucose kinase to convert glucose, a degradation product of starch and / or its derivatives, into glucose 6-phosphate by adding polyphosphate.

[0335] When the carbohydrate is maltose and / or maltooligosaccharide, the enzyme for converting the carbohydrate into β-glucose 1-phosphate comprises maltose phosphorylase, a microorganism expressing maltose phosphorylase, and / or a culture of a microorganism expressing maltose phosphorylase. The β-glucose 1-phosphate is further converted into glucose 6-phosphate by β-phosphoglucose mutase (β-PGM), a microorganism expressing β-phosphoglucose mutase, and / or a culture of a microorganism expressing β-phosphoglucose mutase.

[0336] When the sugar is sucrose, the enzyme for converting the sugar into glucose 1-phosphate comprises sucrose phosphorylase, a microorganism expressing sucrose phosphorylase and / or a culture of a microorganism expressing sucrose phosphorylase. In some other embodiments of the present disclosure, in order to increase the output of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate from the more excellent boron-containing composition may further include converting the degradation product fructose of sucrose into glucose using glucose isomerase (GI), a microorganism expressing glucose isomerase and / or a culture of a microorganism expressing glucose isomerase. Glucose and polyphosphate are then catalyzed by polyphosphate glucokinase to generate glucose 1-phosphate.

[0337] When the sugar is cellulose and / or its derivatives, the cellulose and / or its derivatives can be selected from non-edible lignocellulosic materials (e.g., cellulose, hemicellulose and / or lignin and other minor components), pure cellulose (Avicel (microcrystalline cellulose), regenerated amorphous cellulose, bacterial cellulose, filter paper, etc.), partially hydrolyzed cellulose substrates (including water-insoluble cellodextrins with a degree of polymerization greater than 7, water-soluble cellodextrins with a degree of polymerization of 3-6, cellobiose, glucose and fructose). In certain embodiments of the present disclosure, when the sugar is cellulose and its derivatives, the enzymes used to convert the sugar into glucose 1-phosphate include cellodextrin phosphorylase (CDP), a microorganism expressing cellodextrin phosphorylase and / or a culture of a microorganism expressing cellodextrin phosphorylase, and cellobiose phosphorylase (CBP), a microorganism expressing cellobiose phosphorylase and / or a culture of a microorganism expressing cellobiose phosphorylase.

[0338] Some methods of the present disclosure may further include the step of converting cellulose into a cellulose derivative, for example, using endoglucanase, a microorganism expressing endoglucanase and / or a culture of a microorganism expressing endoglucanase and / or a cellobiohydrolase, a microorganism expressing cellobiohydrolase and / or a culture of a microorganism expressing cellobiohydrolase to hydrolyze solid cellulose into water-soluble cellodextrins and cellobiose. In some embodiments, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include pre-treating cellulose before cellulose is hydrolyzed to generate glucose 1-phosphate, to improve their reactivity and reduce the degree of polymerization of cellulose chains. The pretreatment method of the cellulose includes but is not limited to dilute acid pretreatment, lignocellulose fractionation based on cellulose solvent, ammonia fiber expansion, ammonia soaking, ionic liquid treatment, and partial hydrolysis by using concentrated acid (including hydrochloric acid, sulfuric acid, phosphoric acid and combinations thereof).

[0339] In certain embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting cellobiose, a degradation product of cellulose and / or its derivatives, into glucose 1-phosphate and glucose using cellobiose phosphorylase, a microorganism expressing cellobiose phosphorylase, and / or a culture of a microorganism expressing cellobiose phosphorylase. In some further embodiments of the present disclosure, in order to increase the yield of fructose 6-phosphate, the method for preparing fructose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting glucose, a degradation product of cellulose and / or its derivatives, into glucose 6-phosphate by adding polyphosphate using polyphosphate glucokinase (PPGK), a microorganism expressing polyphosphate glucokinase, and / or a culture of a microorganism expressing polyphosphate glucokinase.

[0340] In some specific examples of the present invention, a method for preparing D-psicose 6-phosphate by catalyzing D-glucose 6-phosphate using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0341] (i) preparing a reaction mixture comprising D-glucose 6-phosphate;

[0342] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0343] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate and an isomerase that epimerizes D-fructose 6-phosphate to D-psicose 6-phosphate.

[0344] In the present invention, the isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate includes but is not limited to D-glucose 6-phosphate isomerase (EC 5.3.1.9); the isomerase that epimerizes D-fructose 6-phosphate to D-psicose 6-phosphate includes but is not limited to D-psicose-6-phosphate 3-position epimerase.

[0345] In the present invention, the method for preparing D-psicose 6-phosphate by catalyzing D-glucose 6-phosphate with the boron-containing composition may further include the step of converting glucose 1-phosphate into glucose 6-phosphate by using phosphoglucomutase, a microorganism expressing phosphoglucomutase and / or a culture of a microorganism expressing phosphoglucomutase.

[0346] In the present invention, the method for preparing D-psicose 6-phosphate from D-glucose 6-phosphate by catalyzing the boron-containing composition may further include the step of converting saccharides (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) into glucose 1-phosphate, wherein the step is catalyzed by at least one enzyme, a microorganism expressing the enzyme, and / or a culture of a microorganism expressing the enzyme, and the saccharide may be selected from, but not limited to, starch and / or its derivatives, cellulose and / or its derivatives, fructose, glucose and / or sucrose. According to the method disclosed herein, the one or more enzymes used in the step of converting saccharides into glucose 1-phosphate may be α-glucan phosphorylase, maltose phosphorylase, sucrose phosphorylase, cellodextrin phosphorylase, cellobiose phosphorylase and / or cellulose phosphorylase, and mixtures thereof, and / or microorganisms expressing the above one or more enzymes and mixtures thereof, and / or cultures of microorganisms expressing the above one or more enzymes and mixtures thereof.

[0347] When the carbohydrate is starch and / or its derivatives, the starch and / or its derivatives can be selected from, including but not limited to, amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, maltose, glucose and mixtures thereof. In certain embodiments of the present disclosure, when the carbohydrate is starch and / or its derivatives, including but not limited to amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, the enzyme for converting the carbohydrate into glucose 1-phosphate comprises α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase.

[0348] Some methods of the present disclosure may further include the step of converting starch into starch derivatives, wherein the starch derivatives are prepared by enzymatic hydrolysis of starch and / or acid hydrolysis of starch. In certain embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting starch into dextrins using a starch branching enzyme such as isoamylase (IA), a microorganism expressing isoamylase, and / or a culture of a microorganism expressing isoamylase. In certain embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting starch into dextrins using a starch branching enzyme such as pullulanase (PA), a microorganism expressing pullulanase, and / or a culture of a microorganism expressing pullulanase.

[0349] In some other embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include using 4-glucan transferase (4GT), a microorganism expressing 4-glucan transferase, and / or a culture of a microorganism expressing 4-glucan transferase to convert the degradation products of starch and / or its derivatives, glucose, maltose, and maltotriose, into longer maltooligosaccharides, wherein the longer maltooligosaccharides can be converted into glucose 1-phosphate using α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase, and / or a culture of a microorganism expressing α-glucan phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include using polyphosphate glucokinase (PPGK), a microorganism expressing polyphosphate glucokinase and / or a culture of a microorganism expressing polyphosphate glucokinase to convert glucose, a degradation product of starch and / or its derivatives, into glucose 6-phosphate by adding polyphosphate.

[0350] When the carbohydrate is maltose and / or maltooligosaccharide, the enzyme for converting the carbohydrate into β-glucose 1-phosphate comprises maltose phosphorylase, a microorganism expressing maltose phosphorylase, and / or a culture of a microorganism expressing maltose phosphorylase. The β-glucose 1-phosphate is further converted into glucose 6-phosphate by β-phosphoglucose mutase (β-PGM), a microorganism expressing β-phosphoglucose mutase, and / or a culture of a microorganism expressing β-phosphoglucose mutase.

[0351] When the sugar is sucrose, the enzyme for converting the sugar into glucose 1-phosphate comprises sucrose phosphorylase, a microorganism expressing sucrose phosphorylase, and / or a culture of a microorganism expressing sucrose phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting the degradation product of sucrose, fructose, into glucose using glucose isomerase (GI), a microorganism expressing glucose isomerase, and / or a culture of a microorganism expressing glucose isomerase. Glucose and polyphosphate are then catalyzed by polyphosphate glucokinase to generate glucose 1-phosphate.

[0352] When the sugar is cellulose and / or its derivatives, the cellulose and / or its derivatives can be selected from non-edible lignocellulosic materials (e.g., cellulose, hemicellulose and / or lignin and other minor components), pure cellulose (Avicel (microcrystalline cellulose), regenerated amorphous cellulose, bacterial cellulose, filter paper, etc.), partially hydrolyzed cellulose substrates (including water-insoluble cellodextrins with a degree of polymerization greater than 7, water-soluble cellodextrins with a degree of polymerization of 3-6, cellobiose, glucose and fructose). In certain embodiments of the present disclosure, when the sugar is cellulose and its derivatives, the enzymes used to convert the sugar into glucose 1-phosphate include cellodextrin phosphorylase (CDP), a microorganism expressing cellodextrin phosphorylase and / or a culture of a microorganism expressing cellodextrin phosphorylase, and cellobiose phosphorylase (CBP), a microorganism expressing cellobiose phosphorylase and / or a culture of a microorganism expressing cellobiose phosphorylase.

[0353] Some methods of the present disclosure may further include the step of converting cellulose into a cellulose derivative, for example, using endoglucanase, a microorganism expressing endoglucanase and / or a culture of a microorganism expressing endoglucanase and / or a cellobiohydrolase, a microorganism expressing cellobiohydrolase and / or a culture of a microorganism expressing cellobiohydrolase to hydrolyze solid cellulose into water-soluble cellodextrins and cellobiose. In some embodiments, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include pre-treating cellulose to increase their reactivity and reduce the degree of polymerization of cellulose chains before cellulose is hydrolyzed to generate glucose 1-phosphate. The pretreatment method of the cellulose includes but is not limited to dilute acid pretreatment, lignocellulose fractionation based on cellulose solvent, ammonia fiber expansion, ammonia soaking, ionic liquid treatment, and partial hydrolysis by using concentrated acid (including hydrochloric acid, sulfuric acid, phosphoric acid and combinations thereof).

[0354] In certain embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting cellobiose, a degradation product of cellulose and / or its derivatives, into glucose 1-phosphate and glucose using cellobiose phosphorylase, a microorganism expressing cellobiose phosphorylase, and / or a culture of a microorganism expressing cellobiose phosphorylase. In still other embodiments of the present disclosure, in order to increase the yield of psicose 6-phosphate, the method for preparing psicose 6-phosphate by catalyzing glucose 6-phosphate with a more excellent boron-containing composition may further include converting glucose, a degradation product of cellulose and / or its derivatives, into glucose 6-phosphate by adding polyphosphate using polyphosphate glucokinase (PPGK), a microorganism expressing polyphosphate glucokinase, and / or a culture of a microorganism expressing polyphosphate glucokinase.

[0355] In some embodiments of the present invention, a method for preparing monosaccharides from glucose 6-phosphate via phosphate monosaccharides is provided using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention, the method comprising the steps of:

[0356] (i) preparing a reaction mixture comprising glucose 6-phosphate;

[0357] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst and a dephosphoenzyme or a phosphatase under suitable reaction conditions;

[0358] The component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes glucose 6-phosphate into other monosaccharide 6-phosphates.

[0359] In some specific examples of the present invention, a method for preparing D-fructose by catalyzing D-glucose 6-phosphate via phosphate monosaccharides using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0360] (i) preparing a reaction mixture comprising D-glucose 6-phosphate;

[0361] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst and a dephosphoenzyme or a phosphatase under suitable reaction conditions;

[0362] The enzyme component (c) in the boron-containing composition or boron-containing composition catalyst includes an isomerase that isomerizes D-glucose 6-phosphate to D-fructose 6-phosphate, and the dephosphorylase or phosphatase is D-fructose 6-phosphate phosphatase.

[0363] In some specific examples of the present invention, a method for preparing D-psicose by catalyzing D-glucose 6-phosphate via phosphate monosaccharides using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0364] (i) preparing a reaction mixture comprising glucose 6-phosphate;

[0365] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst and a dephosphoenzyme or a phosphatase under suitable reaction conditions;

[0366] The component (c) enzyme in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate and an isomerase that epimerizes D-fructose 6-phosphate to D-psicose 6-phosphate, and the dephosphorylase or phosphatase is D-psicose 6-phosphate phosphatase.

[0367] In some specific examples of the present invention, a method for preparing fructose-1,6-diphosphate by catalyzing D-glucose-6-phosphate via phosphate monosaccharide using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0368] (i) preparing a reaction mixture comprising glucose 6-phosphate;

[0369] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst and phosphofructokinase under suitable reaction conditions;

[0370] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes glucose 6-phosphate into fructose 6-phosphate.

[0371] In the present invention, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via monosaccharide phosphates using the boron-containing composition may also include the step of converting glucose 1-phosphate into glucose 6-phosphate by utilizing phosphoglucomutase, a microorganism expressing phosphoglucomutase, and / or a culture of a microorganism expressing phosphoglucomutase.

[0372] In the present invention, the method for preparing monosaccharides and derivatives from glucose 6-phosphate by catalyzing the boron-containing composition via monosaccharide phosphate may further include the step of converting saccharides (such as polysaccharides, oligosaccharides, disaccharides, monosaccharides) into glucose 1-phosphate, wherein the step is catalyzed by at least one enzyme, a microorganism expressing the enzyme, and / or a culture of a microorganism expressing the enzyme, and the saccharide may be selected from, but not limited to, starch and / or its derivatives, cellulose and / or its derivatives, fructose, glucose and / or sucrose. According to the method disclosed herein, the one or more enzymes used in the step of converting saccharides into glucose 1-phosphate may be α-glucan phosphorylase, maltose phosphorylase, sucrose phosphorylase, cellodextrin phosphorylase, cellobiose phosphorylase and / or cellulose phosphorylase, and mixtures thereof, and / or microorganisms expressing the above one or more enzymes and mixtures thereof, and / or cultures of microorganisms expressing the above one or more enzymes and mixtures thereof.

[0373] When the carbohydrate is starch and / or its derivatives, the starch and / or its derivatives can be selected from, including but not limited to, amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, maltose, glucose and mixtures thereof. In certain embodiments of the present disclosure, when the carbohydrate is starch and / or its derivatives, including but not limited to amylose, amylopectin, soluble starch, amylopectin, maltodextrin, maltooligosaccharide, the enzyme for converting the carbohydrate into glucose 1-phosphate comprises α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase.

[0374] Some methods of the present disclosure may further include the step of converting starch into starch derivatives, wherein the starch derivatives are prepared by enzymatic hydrolysis of starch and / or acid hydrolysis of starch. In certain embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives from glucose 6-phosphate via phosphate monosaccharides by the more excellent boron-containing composition may further include converting starch into dextrins by using starch branching enzymes such as isoamylase (IA), microorganisms expressing isoamylases and / or cultures of microorganisms expressing isoamylases. In certain embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives from glucose 6-phosphate via phosphate monosaccharides by the more excellent boron-containing composition may further include converting starch into dextrins by using starch branching enzymes such as pullulanase (PA), microorganisms expressing pullulanase and / or cultures of microorganisms expressing pullulanase.

[0375] In some other embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via phosphate monosaccharides using the more excellent boron-containing composition may further include using 4-glucan transferase (4GT), a microorganism expressing 4-glucan transferase and / or a culture of a microorganism expressing 4-glucan transferase to convert the degradation products of starch and / or its derivatives, glucose, maltose and maltotriose, into longer maltooligosaccharides, wherein the longer maltooligosaccharides can be converted into glucose 1-phosphate using α-glucan phosphorylase, a microorganism expressing α-glucan phosphorylase and / or a culture of a microorganism expressing α-glucan phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via phosphate monosaccharides using a more excellent boron-containing composition may further include using polyphosphate glucose kinase (PPGK), a microorganism expressing polyphosphate glucose kinase and / or a culture of a microorganism expressing polyphosphate glucose kinase to convert glucose, a degradation product of starch and / or its derivatives, into glucose 6-phosphate by adding polyphosphate.

[0376] When the carbohydrate is maltose and / or maltooligosaccharide, the enzyme for converting the carbohydrate into β-glucose 1-phosphate comprises maltose phosphorylase, a microorganism expressing maltose phosphorylase, and / or a culture of a microorganism expressing maltose phosphorylase. The β-glucose 1-phosphate is further converted into glucose 6-phosphate by β-phosphoglucose mutase (β-PGM), a microorganism expressing β-phosphoglucose mutase, and / or a culture of a microorganism expressing β-phosphoglucose mutase.

[0377] When the sugar is sucrose, the enzyme for converting the sugar into glucose 1-phosphate includes sucrose phosphorylase, a microorganism expressing sucrose phosphorylase, and / or a culture of a microorganism expressing sucrose phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives from the more excellent boron-containing composition catalyzing glucose 6-phosphate via phosphate monosaccharides may further include utilizing glucose isomerase (GI), a microorganism expressing glucose isomerase, and / or a culture of a microorganism expressing glucose isomerase to convert the degradation product fructose of sucrose into glucose. Glucose and polyphosphate are then catalyzed by polyphosphate glucokinase to generate glucose 1-phosphate.

[0378] When the sugar is cellulose and / or its derivatives, the cellulose and / or its derivatives can be selected from non-edible lignocellulosic materials (e.g., cellulose, hemicellulose and / or lignin and other minor components), pure cellulose (Avicel (microcrystalline cellulose), regenerated amorphous cellulose, bacterial cellulose, filter paper, etc.), partially hydrolyzed cellulose substrates (including water-insoluble cellodextrins with a degree of polymerization greater than 7, water-soluble cellodextrins with a degree of polymerization of 3-6, cellobiose, glucose and fructose). In certain embodiments of the present disclosure, when the sugar is cellulose and its derivatives, the enzymes used to convert the sugar into glucose 1-phosphate include cellodextrin phosphorylase (CDP), a microorganism expressing cellodextrin phosphorylase and / or a culture of a microorganism expressing cellodextrin phosphorylase, and cellobiose phosphorylase (CBP), a microorganism expressing cellobiose phosphorylase and / or a culture of a microorganism expressing cellobiose phosphorylase.

[0379] Some methods of the present disclosure may further include the step of converting cellulose into cellulose derivatives, for example, using endoglucanase, a microorganism expressing endoglucanase and / or a culture of a microorganism expressing endoglucanase and / or a cellobiohydrolase, a microorganism expressing cellobiohydrolase and / or a culture of a microorganism expressing cellobiohydrolase to hydrolyze solid cellulose into water-soluble cellodextrins and cellobiose. In some embodiments, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via phosphate monosaccharides by the more excellent boron-containing composition may further include pre-treating cellulose to increase their reactivity and reduce the degree of polymerization of cellulose chains before cellulose hydrolysis to generate glucose 1-phosphate. The pretreatment method of the cellulose includes but is not limited to dilute acid pretreatment, lignocellulose fractionation based on cellulose solvent, ammonia fiber expansion, ammonia soaking, ionic liquid treatment, and partial hydrolysis by using concentrated acid (including hydrochloric acid, sulfuric acid, phosphoric acid and combinations thereof).

[0380] In certain embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via phosphate monosaccharides by the more excellent boron-containing composition may further include converting the degradation product of cellulose and / or its derivatives, cellobiose, into glucose 1-phosphate and glucose by using cellobiose phosphorylase, a microorganism expressing cellobiose phosphorylase, and / or a culture of a microorganism expressing cellobiose phosphorylase. In some other embodiments of the present disclosure, in order to increase the yield of monosaccharides and derivatives, the method for preparing monosaccharides and derivatives by catalyzing glucose 6-phosphate via phosphate monosaccharides by the more excellent boron-containing composition may further include converting the degradation product of cellulose and / or its derivatives, glucose, into glucose 6-phosphate by adding polyphosphate using polyphosphate glucokinase (PPGK), a microorganism expressing polyphosphate glucokinase, and / or a culture of a microorganism expressing polyphosphate glucokinase.

[0381] In some embodiments of the present invention, a method for catalytically preparing deoxy sugar using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0382] (i) preparing a reaction mixture comprising a substrate;

[0383] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions;

[0384] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one deoxysugar into other deoxysugars.

[0385] In some specific examples of the present invention, a method for preparing L-rhamnulose (6-deoxyL-fructose) by catalyzing L-rhamnose (6-deoxyL-mannose) using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0386] (i) preparing a reaction mixture comprising L-rhamnose;

[0387] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0388] The enzyme component (c) in the boron-containing composition or boron-containing composition catalyst includes an isomerase that isomerizes L-rhamnose (6-deoxy L-mannose) to L-rhamnulose (6-deoxy L-fructose).

[0389] In the present invention, the isomerase that isomerizes L-rhamnose (6-deoxy L-mannose) to L-rhamnulose (6-deoxy L-fructose) includes but is not limited to L-rhamnose isomerase (EC5.3.1.14).

[0390] In some specific examples of the present invention, a method for preparing 6-deoxy-L-tagatose by catalyzing 6-deoxy-L-galactose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0391] (i) preparing a reaction mixture comprising 6-deoxyL-galactose;

[0392] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0393] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes 6-deoxyL-galactose into 6-deoxyL-tagatose.

[0394] In the present invention, the isomerase that isomerizes 6-deoxyL-galactose to 6-deoxyL-tagatose includes but is not limited to D-arabinose isomerase (EC5.3.1.3).

[0395] In some specific examples of the present invention, a method for preparing 6-deoxy-L-psicose by catalyzing 6-deoxy-L-fructose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0396] (i) preparing a reaction mixture comprising 6-deoxyL-fructose;

[0397] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0398] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes 6-deoxyL-fructose into 6-deoxyL-psicose.

[0399] In the present invention, the isomerase that epimerizes 6-deoxyL-fructose to 6-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0400] In some specific examples of the present invention, a method for preparing 1-deoxy-L-psicose by catalyzing 1-deoxy-L-fructose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0401] (i) preparing a reaction mixture comprising 1-deoxyL-fructose;

[0402] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0403] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes 1-deoxy-L-fructose into 1-deoxy-L-psicose.

[0404] In the present invention, the isomerase that epimerizes 1-deoxyL-fructose to 1-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0405] In some specific examples of the present invention, a method for preparing 6-deoxy-L-psicose by catalyzing L-rhamnose (6-deoxy-L-mannose) using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0406] (i) preparing a reaction mixture comprising L-rhamnose;

[0407] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0408] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes an isomerase that isomerizes L-rhamnose (6-deoxyL-mannose) into L-rhamnulose (6-deoxyL-fructose) and an isomerase that isomerizes 6-deoxyL-fructose into 6-deoxyL-psicose.

[0409] In the present invention, the isomerase that isomerizes L-rhamnose (6-deoxyL-mannose) to L-rhamnulose (6-deoxyL-fructose) includes but is not limited to L-rhamnose isomerase (EC5.3.1.14); the isomerase that epimerizes 6-deoxyL-fructose to 6-deoxyL-psicose includes but is not limited to D-tagatose 3-epimerase (EC 5.1.3.31).

[0410] In some embodiments of the present invention, a method for catalyzing the preparation of disaccharides using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0411] (i) preparing a reaction mixture comprising a substrate;

[0412] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions;

[0413] The enzyme component (c) in the boron-containing composition or the boron-containing composition catalyst includes at least one isomerase that isomerizes one disaccharide into other disaccharides.

[0414] In some specific examples of the present invention, a method for preparing lactulose by catalyzing lactose using the boron-containing composition of the first aspect of the present invention or the boron-containing composition prepared by the second aspect of the present invention is provided, the method comprising the steps of:

[0415] (i) preparing a reaction mixture comprising lactose;

[0416] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions;

[0417] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes lactose into lactulose.

[0418] In the present invention, lactose refers to a lactose-containing component, including α-lactose monohydrate, β-lactose monohydrate, anhydrous α-lactose, anhydrous β-lactose and amorphous lactose.

[0419] In the present invention, the isomerase that isomerizes lactose into lactulose includes but is not limited to cellobiose-2-epimerase (EC 5.1.3.11).

[0420] In some specific examples of the present invention, a method for preparing turanoose from sucrose by catalyzing the boron-containing composition prepared according to the first aspect of the present invention or the boron-containing composition prepared according to the second aspect of the present invention is provided, the method comprising the steps of:

[0421] (i) preparing a reaction mixture comprising sucrose;

[0422] (ii) contacting the reaction mixture with a boron-containing composition or a boron-containing composition catalyst under suitable reaction conditions.

[0423] The enzyme as component (c) in the boron-containing composition or the boron-containing composition catalyst comprises an isomerase that isomerizes sucrose into turanoose.

[0424] In the present invention, the isomerase that isomerizes sucrose to turanoose includes but is not limited to amylosucrase (EC 2.4.1.4).

[0425] Example

[0426] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0427] Materials and Methods

[0428] Drugs and materials:

[0429] Unless otherwise noted, all drugs were of analytical grade or higher and were purchased from Sigma-Aldrich (St. Louis, MO, USA) or Sinopharm (Shanghai, China). E. coli TOP10 and DH5α (Thermo Fisher Scientific, Waltham, MA, USA) were used for DNA manipulation and plasmid amplification. E. coli BL21 (DE3) (Invitrogen Life Sciences, Carlsbad, CA, USA) was used for recombinant protein expression.

[0430] Preparation and purification of plasmids and recombinant proteins

[0431] A person skilled in the art can prepare recombinant engineered bacteria using the known techniques in the art according to their actual needs, and can refer to Molecular Cloning Experiment Guide (3rd Edition) (Science Press). The vectors, plasmids, and hosts used in the experiment are all conventional vectors, plasmids, and host series used for expression of bacteria (e.g., E. coli), such as PET series vectors, plasmids, and BL21 series host bacteria; the culture medium used is a culture medium for conventional bacteria (e.g., E. coli) engineered bacteria, such as LB culture medium; the culture method used is a conventional culture method for bacteria (e.g., E. coli) engineered bacteria.

[0432] Exemplarily, the preparation method of the pET plasmid carrying the corresponding protein encoding gene is as follows.

[0433] The genes of proteins with corresponding functions are amplified from their corresponding genomes or obtained by gene synthesis. They are inserted into pET series plasmids through simple cloning technology based on extended overlap extension PCR (POE-PCR) (You, C., X.-Z. Zhang and Y.-HP Zhang (2012). "Simple Cloning: direct transformation of PCR product (DNA multimer) to Escherichia coli and Bacillus subtilis." Appl. Environ. Microbiol. 78: 1593-1595.), and plasmids containing nucleotide sequences encoding corresponding target proteins are constructed.

[0434] Transform the plasmid containing the nucleotide sequence encoding the corresponding target protein into E. coli BL21 (DE3) competent cells, pick a single colony from overnight culture and add it to a 5mL LB test tube containing the corresponding resistance, and culture it overnight at 37℃ constant temperature shaker 200rpm. Inoculate it into a triangular flask containing the corresponding resistance and 200mL LB medium according to the inoculum of 1% (v / v), and expand the culture in a 37℃ constant temperature shaker 200rpm. When OD 600 When the concentration reaches 0.8-1.0, add the inducer IPTG to a final concentration of 0.1mM, and culture at 200rpm in a 16°C constant temperature shaker for 16-18h to express the target protein and obtain a cell culture fluid. The cell culture fluid is centrifuged at 4°C to collect cells, and the collected cells are washed once with a 0.9% (w / v) sodium chloride solution. The obtained cells can be used as a source of component c enzyme after heat treatment.

[0435] The cell pellet washed once with the sodium chloride solution is resuspended in a 50 mM HEPES (pH 7.0) buffer solution and lysed by ultrasonication to obtain a cell lysate, and after centrifugation, the target protein in the supernatant is purified. The obtained protein can also be used as a source of the enzyme of component c.

[0436] The component (c) enzyme included in the boron-containing composition of the present invention is usually represented by the reaction (i.e., by specificity and function) based on their catalysis. The enzyme used in the following examples can be easily found in journal literature or related databases (NCBI, KEGG, BRENDA, EMBL, etc.) through the information in Table 1.

[0437] Table 1

[0438]

[0439]

[0440] In each of the following examples, the enzyme activity units used can be increased or decreased according to actual needs so that the component (c) enzyme is kept within the weight range of the present invention. The activity units and weights of the enzyme can be simply converted according to actual needs. For example, if the activity of the corresponding functional enzyme used is low, the amount of the component (c) enzyme can be increased in order to reduce the reaction time and maintain the reaction rate.

[0441] Example 1 A composition containing zinc orthoborate, isomerase for isomerizing D-glucose to D-fructose, and alginic acid to catalyze D-Glucose to D-Fructose

[0442] A composition containing zinc orthoborate comprises 50 parts by weight of calcium alginate, 10 parts by weight of diatomaceous earth, 35 parts by weight of zinc orthoborate, and 5 parts by weight of isomerase for isomerizing D-glucose into D-fructose. The preparation method thereof is as follows:

[0443] 35 mg of zinc orthoborate and 10 mg of diatomaceous earth were dispersed in 10 mL of 5 g / L sodium alginate solution, 5 mg of isomerase for isomerizing D-glucose to D-fructose was added, and the mixture was evenly mixed. The solution was ultrasonically degassed and added dropwise to a 4 g / L calcium chloride solution. After incubation for 6 hours, the calcium alginate gel balls were filtered and washed with deionized water for 3 times to obtain a composition containing zinc orthoborate and isomerase for isomerizing D-glucose to D-fructose.

[0444] The boron-containing composition is used to catalyze D-glucose to prepare D-fructose, and the method is as follows:

[0445] In a 1mL preparation system, 100mM HEPES buffer (pH 7.5), 10mM magnesium sulfate, 278mM D-glucose, 5.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 55°C. The amount of D-fructose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was BioRad 87H, the mobile phase was 5mM sulfuric acid, the flow rate was 0.6mL / min, the column temperature was 60°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-fructose produced was 164mM and the conversion rate was 59.0%.

[0446] Comparative Example 1

[0447] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.5), 10 mM magnesium sulfate, 278 mM D-glucose, and 0.25 g / L D-glucose isomerase are reacted at 55°C. The amount of D-fructose generated is detected by high performance liquid chromatography. The detection method is as shown in Example 1. HPLC detection shows that the amount of D-fructose generated is 135.6 mM and the conversion rate is 48.8%.

[0448] HPLC test results are as follows Figure 4 The experimental results show that, compared with Comparative Example 1, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-fructose to the substrate D-glucose in Example 1 is significantly increased.

[0449] Example 2 A method comprising aminophenylboronic acid, an isomerase for isomerizing D-xylose to D-xylulose, an amino resin, and a magnetic Nanoparticle composition catalyzes the production of D-xylulose from D-xylose

[0450] A composition containing aminophenylboronic acid comprises 6 parts by weight of aminophenylboronic acid, 36 parts by weight of amino resin, 48 parts by weight of magnetic nanoparticles, and 9 parts by weight of isomerase for isomerizing D-xylose to D-xylulose. The preparation method thereof is as follows:

[0451] 3.6 g of amino resin was dispersed in 50 mL of 25% glutaraldehyde solution, stirred at room temperature for 24 h, filtered to collect the resin, washed 3 times with deionized water, and then the glutaraldehyde-modified resin was collected. Then, the glutaraldehyde-modified resin was dispersed in a solution containing 0.6 g of 3-aminophenylboronic acid, incubated for 24 h, filtered to collect the resin, and washed with deionized water to obtain a resin component containing aminophenylboronic acid.

[0452] Prepare 50mM Tris-HCl (pH 7.5) solution, take 100ml Tris-HCl solution, add 200mg dopamine solution to fully dissolve, then disperse 4.8g magnetic ferroferric oxide nanoparticles in the above solution, and stir mechanically for 24h. Collect ferroferric oxide by magnet, and collect ferroferric oxide after repeated washing with deionized water. Add 250mL 5g / L D-xylose isomerase to D-xylulose isomerase to polydopamine-modified ferroferric oxide nanoparticles and react for 6h. The residual enzyme concentration in the supernatant was determined by Bradford method, and the weight ratio of enzyme to ferroferric oxide was calculated by subtraction method to be 9:48.

[0453] The resin component containing aminophenylboronic acid and the ferroferric oxide component containing isomerase for isomerizing D-galactose into D-tagatose are mixed according to a certain ratio to obtain a composition containing aminophenylboronic acid and isomerase for isomerizing D-galactose into D-tagatose.

[0454] The boron-containing composition is used to catalyze D-xylose to prepare D-xylulose, and the method is as follows:

[0455] In a 1mL preparation system, 100mM HEPES buffer (pH 8.0), 10mM magnesium sulfate, 333mM D-xylose, 1.5g / L of the boron-containing composition prepared above were reacted at a constant temperature of 55°C. The amount of D-xylulose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was BioRad 87H, the mobile phase was 5mM sulfuric acid, the flow rate was 0.6mL / min, the column temperature was 60°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-xylulose produced was 172mM and the conversion rate was 51.7%.

[0456] Comparative Example 2

[0457] In 1 mL of the preparation system, 100 mM HEPES buffer (pH 8.0), 10 mM magnesium sulfate, 333 mM D-xylose, 0.135 g / L D-xylose isomerase isomerized to D-xylulose, and the reaction is carried out at a constant temperature of 55°C. The amount of D-xylulose produced is detected by high performance liquid chromatography. The detection method is shown in Example 2. HPLC detection shows that the amount of D-xylulose produced is 7.6 mM and the conversion rate is 22.8%.

[0458] HPLC test results are as follows Figure 5 The experimental results show that, compared with Comparative Example 2, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-xylulose to the substrate D-xylose in Example 2 is significantly increased.

[0459] Example 3 Catalysis of a composition containing boric acid, isomerase for isomerization of D-galactose to D-tagatose, and polyvinyl alcohol Preparation of D-Tagatose from D-Galactose

[0460] A boric acid-containing composition comprises 70 parts by weight of polyvinyl alcohol, 20 parts by weight of boric acid, and 10 parts by weight of isomerase for isomerizing D-galactose to D-tagatose. The preparation method thereof is as follows:

[0461] Dissolve polyvinyl alcohol in water to prepare a solution, then add a saturated boric acid solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to boric acid being 2.5:1, stir well and let stand for 12 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing boric acid.

[0462] 20 g of polyvinyl alcohol was dissolved in 1 L of water, and then 10 g of isomerase for isomerizing D-galactose to D-tagatose was added to the polyvinyl alcohol solution, and the mixture was stirred for 30 min. Then, the aqueous solvent was evaporated at room temperature to obtain a polyvinyl alcohol film containing isomerase for isomerizing D-galactose to D-tagatose.

[0463] The polyvinyl alcohol component containing boric acid and the polyvinyl alcohol film containing isomerase for isomerizing D-galactose to D-tagatose are mixed according to a certain ratio to obtain a composition containing boric acid and isomerase for isomerizing D-galactose to D-tagatose.

[0464] The boron-containing composition is used to catalyze D-galactose to prepare D-tagatose, and the method is as follows:

[0465] In a 1mL preparation system, 100mM HEPES buffer (pH 7.2), 10mM magnesium sulfate, 278mM D-galactose, 5.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 65°C. The amount of D-tagatose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-tagatose produced was 172.8mM and the conversion rate was 62.2%.

[0466] Comparative Example 3

[0467] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.2), 10 mM magnesium sulfate, 278 mM D-galactose, 0.5 g / L D-galactose isomerase isomerized to D-tagatose, and the reaction is carried out at a constant temperature of 65 ° C. The amount of D-tagatose produced is detected by high performance liquid chromatography. The detection method is shown in Example 3. HPLC detection shows that the amount of D-tagatose produced is 8.4 mM and the conversion rate is 30.2%.

[0468] The experimental results show that compared with Comparative Example 3, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-tagatose to the substrate D-galactose in Example 3 is significantly increased.

[0469] Example 4 A method for isomerizing D-fructose into D-alcohol containing carboxyl phenylboronic acid, magnetic ferroferric oxide, silicon oxide Combination of ketose epimerases catalyzes the production of D-psicose from D-fructose

[0470] A composition containing carboxyphenylboronic acid comprises 5 parts by weight of carboxyphenylboronic acid, 57 parts by weight of magnetic ferrosoferric oxide, 30 parts by weight of inorganic silicon oxide particles, and 8 parts by weight of epimerase for isomerizing D-fructose into D-psicose. The preparation method thereof is as follows:

[0471] Prepare 50mM Tris-HCl (pH 8.5) solution, take 100mL Tris-HCl solution, add 200mg dopamine solution to fully dissolve, then disperse 5.2g magnetic ferroferric oxide nanoparticles in the above solution, and stir mechanically for 12h. Collect ferroferric oxide by magnet, and collect ferroferric oxide after repeated washing with deionized water. Add 100mL anhydrous methanol to the ferroferric oxide nanoparticles modified with polydopamine, and then add 1g carboxyphenylboric acid and react for 24h. Finally, collect ferroferric oxide modified with carboxyphenylboric acid by magnet to obtain ferroferric oxide component containing carboxyphenylboric acid.

[0472] The epimerase for isomerizing D-fructose to D-psicose was dissolved in 20 mM phosphate buffer (pH 7.0) to a final concentration of 1 g / L. 8 mL of the enzyme solution was added with 2 mL of 1 g / L polyethyleneimine with a molecular weight of 1800, and then 298 mg of tetraethyl orthosilicate (TEOS), and the mixture was stirred for 20 min. After centrifugation and washing, the precipitate was collected to obtain a silicon oxide component containing the epimerase for isomerizing D-fructose to D-psicose.

[0473] The ferrositol component containing carboxyphenylboronic acid and the silicon oxide component containing the epimerase for isomerizing D-fructose into D-psicose are mixed according to a certain ratio to obtain a composition containing carboxyphenylboronic acid and the epimerase for isomerizing D-fructose into D-psicose.

[0474] The boron-containing composition is used to catalyze D-fructose to prepare D-psicose, and the method is as follows:

[0475] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.2), 10 mM magnesium sulfate, 1 mM cobalt chloride, 278 mM D-fructose, and 2.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 60°C. The amount of D-psicose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5 mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-psicose produced was 180.6 mM and the conversion rate was 64.9%.

[0476] Comparative Example 4

[0477] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.2), 10 mM magnesium sulfate, 1 mM cobalt chloride, 278 mM D-fructose, and 0.16 g / L epimerase for isomerizing D-fructose to D-psicose were reacted at a constant temperature of 60°C. The amount of D-psicose produced was detected by high performance liquid chromatography. The detection method is shown in Example 4. HPLC detection showed that the amount of D-psicose produced was 82.8 mM and the conversion rate was 29.8%.

[0478] HPLC test results are as follows Figure 6 The experimental results show that compared with Comparative Example 4, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-psicose to the substrate D-fructose in Example 4 is significantly increased.

[0479] Example 5 A method comprising magnesium metaborate, isomerase for converting D-glucose into D-fructose, and isomerase for converting D-fructose into D-allo Preparation of D-psicose from D-glucose catalyzed by a combination of ketose epimerase and calcium alginate

[0480] A composition containing magnesium metaborate comprises 50 parts by weight of calcium alginate, 5 parts by weight of silicon oxide, 25 parts by weight of magnesium metaborate, 13 parts by weight of isomerase for isomerizing D-glucose into D-fructose, and 7 parts by weight of epimerase for isomerizing D-fructose into D-psicose. The preparation method thereof is as follows:

[0481] 25 mg of magnesium metaborate and 5 mg of silicon oxide are dispersed in 10 mL of 5 g / L sodium alginate solution, 13 mg of isomerase from D-glucose isomerizes to D-fructose and 7 mg of epimerase from D-fructose isomerizes to D-psicose are added and mixed evenly, the above solution is ultrasonically degassed, and added dropwise to 4 g / L calcium chloride solution. After incubation for 6 hours, the calcium alginate gel balls are filtered, and washed with deionized water 3 times to obtain a composition containing magnesium metaborate, isomerase from D-glucose isomerizes to D-fructose, and epimerase from D-fructose isomerizes to D-psicose.

[0482] The boron-containing composition is used to catalyze D-glucose to prepare D-psicose, and the method is as follows:

[0483] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.5), 10 mM magnesium sulfate, 1 mM cobalt chloride, 278 mM D-glucose, and 6.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 60°C. The amount of D-psicose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5 mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-psicose produced was 125 mM and the conversion rate was 45.0%.

[0484] Comparative Example 5

[0485] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.5), 10 mM magnesium sulfate, 1 mM cobalt chloride, 278 mM D-glucose, 0.78 g / L D-glucose isomerase for isomerization to D-fructose, and 0.42 g / L D-fructose isomerase for isomerization to D-psicose were reacted at 60°C. The amount of D-psicose produced was detected by high performance liquid chromatography. The liquid phase detection conditions are as shown in Example 5. HPLC detection showed that the amount of D-psicose produced was 42.2 mM and the conversion rate was 15.2%.

[0486] The experimental results show that compared with Comparative Example 5, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-psicose to the substrate D-glucose in Example 5 is significantly increased.

[0487] Example 6 A method comprising calcium metaborate, isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate, calcium alginate, Chitosan composition catalyzes glucose 6-phosphate to produce fructose 6-phosphate

[0488] A composition containing calcium metaborate comprises 40 parts by weight of calcium alginate, 30 parts by weight of chitosan, 25 parts by weight of calcium metaborate, and 5 parts by weight of isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate. The preparation method thereof is as follows:

[0489] Disperse 25 mg of calcium metaborate in 10 mL of 4 g / L sodium alginate solution, mix well, degas the solution by ultrasonication, add dropwise into 3 g / L calcium chloride solution, incubate for 6 h, filter the calcium alginate gel balls, wash with deionized water 3 times, and obtain a calcium alginate component containing calcium metaborate.

[0490] Chitosan is dissolved in an acetic acid solution to prepare a 1.6% chitosan solution, and isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate is dissolved in the chitosan solution to make the concentration of isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate 2%. After ultrasonic degassing, the mixed solution is added dropwise to a 2% NaOH solution to form gel balls, which are repeatedly washed with deionized water to obtain a chitosan component containing isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate.

[0491] The calcium alginate component containing calcium metaborate and the chitosan component containing isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate are mixed to obtain a composition containing calcium borate and isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate.

[0492] The boron-containing composition is used to catalyze glucose 6-phosphate to prepare fructose 6-phosphate, and the method is as follows:

[0493] In a 1mL preparation system, 100mM HEPES buffer (pH 7.5), 10mM magnesium sulfate, 50mM glucose 6-phosphate, 3.0g / L of the boron-containing composition prepared above were reacted at 70°C. After the reaction, phosphatase was added to remove phosphorus from the phosphate compound, and the amount of D-fructose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-fructose produced was 4.5g / L, and the amount of fructose 6-phosphate produced was calculated to be 24mM, and the conversion rate was 48%.

[0494] Comparative Example 6

[0495] In a 1mL preparation system, 100mM HEPES buffer (pH 7.5), 10mM magnesium sulfate, 50mM glucose 6-phosphate, 0.15g / L glucose 6-phosphate isomerase isomerized to fructose 6-phosphate, and the reaction is carried out at 70°C. After the reaction, phosphatase is added to remove phosphorus from the phosphate compound, and the amount of D-fructose generated is detected by high performance liquid chromatography. The liquid phase detection conditions are shown in Example 6. HPLC detection shows that the amount of D-fructose generated is 2.2g / L, and the amount of fructose 6-phosphate generated is calculated to be 12.2mM, and the conversion rate is 24.4%.

[0496] The experimental results show that compared with Comparative Example 6, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product fructose 6-phosphate to the substrate glucose 6-phosphate in Example 6 is significantly increased.

[0497] Example 7 A method comprising calcium metaborate, α-glucan phosphorylase, glucose phosphoglucomutase, glucose 6-phosphate Preparation of fructose 6-phosphate from maltodextrin catalyzed by a composition of isomerase, calcium alginate and diatomaceous earth for isomerizing fructose 6-phosphate

[0498] A composition containing calcium metaborate comprises 50 parts by weight of calcium alginate, 10 parts by weight of diatomaceous earth, 20 parts by weight of calcium metaborate, 10 parts by weight of α-glucan phosphorylase, 5 parts by weight of glucose phosphoglucomutase, and 5 parts by weight of isomerase for isomerizing glucose 6 phosphate to fructose 6 phosphate. The preparation method thereof is as follows:

[0499] 20 mg of calcium metaborate and 10 mg of diatomaceous earth were dispersed in 10 mL of 6 g / L calcium alginate solution, 10 mg of α-glucan phosphorylase, 5 mg of glucose phosphomutase, and 5 mg of glucose 6-phosphate isomerase to fructose 6-phosphate were added, and the mixture was evenly mixed. The above solution was ultrasonically degassed, and the mixture was dropwise added into a 5 g / L calcium chloride solution. After incubation for 6 hours, the calcium alginate gel balls were filtered, and the balls were washed with deionized water for 3 times to obtain a composition containing calcium metaborate and α-glucan phosphorylase, glucose phosphomutase, glucose 6-phosphate isomerase to fructose 6-phosphate, calcium alginate, and diatomaceous earth.

[0500] The boron-containing composition is used to catalyze dextrin to prepare fructose 6-phosphate in the following manner:

[0501] In a 1mL preparation system, 100mM sodium phosphate buffer (pH 7.2), 5mM magnesium sulfate, 55mM glucose equivalent maltodextrin, 5.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 70°C. After the reaction, phosphatase was added to remove phosphorus from the phosphate compound, and the amount of D-fructose generated was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the amount of D-fructose generated was 4.4g / L, and the amount of fructose 6-phosphate generated was calculated to be 24.4mM, and the conversion rate was 44.4%.

[0502] Comparative Example 7

[0503] In a 1mL preparation system, 100mM sodium phosphate buffer (pH 7.2), 5mM magnesium sulfate, 55mM glucose equivalent dextrin, 0.5g / L α-glucan phosphorylase, 0.25g / L glucose phosphomutase, 0.25g / L glucose 6-phosphate isomerase isomerase isomerized to fructose 6-phosphate, and the reaction is carried out at 70°C. After the reaction is completed, phosphatase is added to remove phosphorus from the phosphate compound, and the amount of D-fructose generated is detected by high performance liquid chromatography. The liquid phase detection conditions are shown in Example 7. HPLC detection shows that the amount of D-fructose generated is 1.98g / L, and the amount of fructose 6-phosphate generated is calculated to be 11mM, with a conversion rate of 20%.

[0504] The experimental results show that compared with Comparative Example 7, under the same enzyme dosage, the same reaction and detection conditions, the yield of the product fructose 6-phosphate in Example 7 is significantly increased.

[0505] Example 8 A method comprising sodium borate, glucose 6-phosphate isomerase for isomerizing fructose 6-phosphate, and fructose 6-phosphate isomerase Composition of epimerase constituting psicose 6-phosphate and calcium alginate catalyzes preparation of D-psicone from D-glucose 6-phosphate sugar

[0506] A sodium borate-containing composition comprises 60 parts by weight of calcium alginate, 3 parts by weight of activated carbon, 20 parts by weight of sodium borate, 3 parts by weight of isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate, and 14 parts by weight of epimerase for isomerizing fructose 6-phosphate to psicose 6-phosphate. The preparation method thereof is as follows:

[0507] 20 mg of sodium borate and 3 mg of activated carbon were dispersed in 10 mL of 6 g / L sodium alginate solution, 3 mg of isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate and 14 mg of epimerase that isomerizes fructose 6-phosphate to psicose 6-phosphate were added and mixed evenly, the above solution was ultrasonically degassed, and the solution was dropwise added into a 4 g / L calcium chloride solution. After incubation for 6 hours, the calcium alginate gel balls were filtered, and washed with deionized water for 3 times to obtain a composition containing sodium borate, isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate, and epimerase that isomerizes fructose 6-phosphate to psicose 6-phosphate.

[0508] The boron-containing composition is used to catalyze glucose 6-phosphate to prepare psicose, and the method is as follows:

[0509] In a 1mL preparation system, 100mM HEPES buffer (pH 7.2), 10mM magnesium sulfate, 1mM cobalt chloride, 50mM glucose 6-phosphate, 1.0g / L of the boron-containing composition prepared above, 1g / L psicose 6-phosphatase were reacted at a constant temperature of 50°C. The production of glucose, fructose, and psicose was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5mL / min, the column temperature was 80°C, and the differential refractive index detector was used. HPLC detection showed that the production of glucose was 1mM, the production of fructose was 2mM, and the production of psicose was 47mM.

[0510] Comparative Example 8

[0511] In 1 mL of the preparation system, 100 mM HEPES buffer (pH 7.2), 10 mM magnesium sulfate, 1 mM cobalt chloride, 50 mM glucose 6-phosphate, 0.03 g / L glucose 6-phosphate isomerase for isomerization to fructose 6-phosphate, 0.14 g / L fructose 6-phosphate isomerase for isomerization to psicose 6-phosphate, 1 g / L psicose 6-phosphatase were reacted at 50°C. The production of glucose, fructose and psicose was detected by high performance liquid chromatography. The liquid phase detection conditions are shown in Example 16. The HPLC detection results showed that the production of glucose was 6 mM, the production of fructose was 5 mM, and the production of psicose was 39 mM.

[0512] The experimental results show that compared with Comparative Example 8, under the same enzyme dosage, the same reaction and detection conditions, the production of the product psicose in Example 8 is significantly increased, and the production of by-products glucose and fructose is significantly reduced. The boron-containing composition helps to increase the product yield.

[0513] Example 9 A method comprising aminophenylboronic acid, an isomerase for isomerizing L-rhamnose to L-rhamnulose, and an isomerase for isomerizing L-rhamnulose to L-rhamnulose Composition of epimerase and organosilicon with the structure of 6-deoxy-L-psicose catalyzes the preparation of 6-deoxy-L-psicone from L-rhamnose sugar

[0514] A composition containing aminophenylboronic acid comprises 55 parts by weight of organosilicon, 15 parts by weight of aminophenylboronic acid, 15 parts by weight of isomerase cells for isomerizing L-rhamnose to L-rhamnulose and 15 parts by weight of epimerase cells for isomerizing L-rhamnulose to 6-deoxy-L-psicose. The preparation method thereof is as follows:

[0515] 15 mg of isomerase cells containing L-rhamnose isomerized to L-rhamnulose and 15 mg of epimerase cells containing L-rhamnulose isomerized to 6-deoxy-L-psicose were resuspended in 20 mM phosphate buffer (pH 7.0), 54 mg of TEOS and 1 mg of 3-aminopropyltriethoxysilane (APTES) were added thereto, and 2% glutaraldehyde solution was added to the above solution after stirring for 2 hours, and stirring was continued for 2 hours. The cells were collected by centrifugation, washed with deionized water 3 times, and then resuspended in 20 mM phosphate buffer (pH 7.0), 15 mg of aminophenylboronic acid was added, and the reaction was continued for 12 hours. After centrifugation and washing, a composition containing aminophenylboronic acid, isomerase for L-rhamnose isomerization to L-rhamnulose and epimerase for L-rhamnulose isomerization to 6-deoxy-L-psicose was obtained.

[0516] The boron-containing composition is used to catalyze L-rhamnose to prepare 6-deoxy-L-psicose, and the method is as follows:

[0517] In a 1 mL preparation system, 100 mM Tris-HCl buffer (pH 8.0), 10 mM magnesium sulfate, 100 mM L-rhamnose, and 5.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 40 ° C. The amount of 6-deoxy L-psicose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Waters Sugar-Pak I, the mobile phase was ultrapure water, the flow rate was 0.5 mL / min, the column temperature was 80 ° C, and the differential refractive index detector was used. HPLC detection showed that the amount of 6-deoxy L-psicose produced was 15 mM and the conversion rate was 15%.

[0518] Comparative Example 9

[0519] In a 1 mL preparation system, 100 mM Tris-HCl buffer (pH 8.0), 10 mM magnesium sulfate, 100 mM L-rhamnose, 0.75 g / L L-rhamnose isomerase cells for isomerizing to L-rhamnulose and 0.75 g / L L-rhamnulose isomerase cells for isomerizing to 6-deoxy-L-psicose were reacted at a constant temperature of 40°C. The amount of 6-deoxy-L-psicose produced was detected by high performance liquid chromatography. The liquid phase detection conditions are as shown in Example 17. The HPLC detection results showed that the amount of 6-deoxy-L-psicose produced was 8 mM and the conversion rate was 8%.

[0520] The experimental results show that compared with Comparative Example 9, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product 6-deoxy-L-psicose to the substrate L-rhamnose in Example 9 is significantly increased.

[0521] Example 10 A composition containing boric acid, an isomerase for isomerizing sucrose to turanose, and polyvinyl alcohol catalyzes the isomerization of sucrose to turanose Turano sugar

[0522] A boric acid-containing composition comprises 60 parts by weight of polyvinyl alcohol, 10 parts by weight of boric acid, and 30 parts by weight of isomerase for isomerizing sucrose to turanose. The preparation method thereof is as follows:

[0523] Dissolve polyvinyl alcohol in water to prepare a solution, then add a saturated boric acid solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to boric acid being 6:1, stir and mix thoroughly and let stand for 24 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing boric acid.

[0524] The polyvinyl alcohol component containing boric acid and the isomerase that isomerizes sucrose to turanoose are mixed according to a certain ratio to obtain a composition containing boric acid and the isomerase that isomerizes sucrose to turanoose.

[0525] The boron-containing composition is used to catalyze sucrose to prepare turanoose, and the method is as follows:

[0526] In a 1mL preparation system, 100mM Tris-HCl buffer (pH 7.5), 2M sucrose, and 10.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 45°C. The amount of turanoose produced was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Thermo Syncronis Amino, the mobile phase was acetonitrile and water (80:20, v / v), the flow rate was 0.8mL / min, the column temperature was 35°C, and the differential refractive index detector was used. HPLC detection showed that the amount of turanoose produced was 1.2M and the conversion rate was 60%.

[0527] Comparative Example 10

[0528] In a 1 mL preparation system, 100 mM Tris-HCl buffer (pH 7.5), 2 M sucrose, and 3.0 g / L sucrose isomerase for isomerization to turanosose were reacted at 45°C. The amount of turanosose produced was detected by high performance liquid chromatography. The liquid phase detection conditions are as shown in Example 10. The HPLC detection results showed that the amount of turanosose produced was 960 mM and the conversion rate was 48%.

[0529] The experimental results show that compared with Comparative Example 10, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product turanoose to the substrate sucrose in Example 10 is significantly increased.

[0530] Example 11 A combination containing zinc orthoborate, an isomerase for isomerizing lactose into lactulose, and calcium alginate Preparation of Lactulose from Lactose Catalyzed by Biocatalysis

[0531] A composition containing zinc orthoborate comprises 60 parts by weight of calcium alginate, 8 parts by weight of diatomaceous earth, 20 parts by weight of zinc orthoborate, and 12 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0532] 20 mg of zinc orthoborate and 8 mg of diatomaceous earth are dispersed in 10 mL of 6 g / L sodium alginate solution, 12 mg of lactose isomerase for isomerizing lactulose is added, and the mixture is evenly mixed. The solution is ultrasonically degassed, and the solution is dropped into a 4 g / L calcium chloride solution. After incubation for 6 hours, the calcium alginate gel balls are filtered, and the balls are washed with deionized water three times to obtain a composition containing zinc orthoborate and lactose isomerase for isomerizing lactulose.

[0533] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0534] In a 1mL preparation system, 100mM HEPES buffer (pH 7.5), 1M lactose, 5.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 60°C. The production of lactulose and ipilactose was detected by high performance liquid chromatography. The chromatographic column used for liquid chromatography was Hypersil GOLD Amino, the mobile phase was acetonitrile-phosphate buffer (1.15g of sodium dihydrogen phosphate was dissolved in 1L of water) (84:16, v / v), the flow rate was 1.2mL / min, the column temperature was 40°C, and the differential refractive index detector was used. HPLC detection showed that when the reaction reached 8h, the production of lactulose was 880mM, the production of ipilactose was 15mM, and the lactulose conversion rate was 88.0%.

[0535] Comparative Example 11

[0536] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.5), 1 M lactose, 0.6 g / L lactose isomerase for isomerization to lactulose were reacted at 60°C. The production of lactulose and epililactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that the production of lactulose was 560 mM and the production of epililactose was 120 mM after 8 h of reaction, and the conversion rate of lactulose was 56.0%.

[0537] The experimental results show that compared with Comparative Example 11, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product lactulose to the substrate lactose in Example 11 increased significantly, and the ratio of the byproduct epiratose to the substrate lactose decreased significantly.

[0538] Example 12 A method of isomerizing lactose into lactulose containing aminophenylboronic acid, an isomerase, an amino resin, and a magnetic Composition of nanoparticles catalyzes the production of lactulose from lactose

[0539] A composition containing aminophenylboronic acid comprises 6 parts by weight of aminophenylboronic acid, 36 parts by weight of amino resin, 48 parts by weight of magnetic nanoparticles, and 9 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0540] 3.6 g of amino resin was dispersed in 50 mL of 25% glutaraldehyde solution, stirred at room temperature for 24 h, filtered to collect the resin, washed 3 times with deionized water, and then the glutaraldehyde-modified resin was collected. Then, the glutaraldehyde-modified resin was dispersed in a solution containing 0.6 g of 3-aminophenylboronic acid, incubated for 24 h, filtered to collect the resin, and washed with deionized water to obtain a resin component containing aminophenylboronic acid.

[0541] Prepare 50mM Tris-HCl (pH 7.5) solution, take 100ml Tris-HCl solution, add 200mg dopamine solution to fully dissolve, then disperse 4.8g magnetic ferroferric oxide nanoparticles in the above solution, and stir mechanically for 24h. Collect ferroferric oxide by magnet, and collect ferroferric oxide after repeated washing with deionized water. Add 250mL 5g / L lactose isomerase for isomerization to lactulose to polydopamine-modified ferroferric oxide nanoparticles and react for 6h. The residual enzyme concentration in the supernatant was determined by Bradford method, and the weight ratio of enzyme to ferroferric oxide was calculated by subtraction method to be 9:48.

[0542] The resin component containing aminophenylboronic acid and the ferroferric oxide component containing the epimerase for isomerizing lactose into lactulose are mixed according to a certain ratio to obtain a composition containing aminophenylboronic acid and the epimerase for isomerizing lactose into lactulose.

[0543] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0544] In a 1 mL preparation system, 100 mM Tris-HCl buffer (pH 8.0), 2 M lactose, and 20.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 45 ° C. The production of lactulose and ipilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 2 hours, the production of lactulose was 1560 mM, the production of ipilactose was 28 mM, and the lactulose conversion rate was 78.0%.

[0545] Example 13 A combination containing borax, an isomerase for isomerizing lactose into lactulose, and polyvinyl alcohol Preparation of Lactulose from Lactose Catalyzed by Biocatalysis

[0546] A composition containing triborax comprises 70 parts by weight of polyvinyl alcohol, 20 parts by weight of triborax, and 10 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0547] Dissolve polyvinyl alcohol in water to prepare a solution, then add borax solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to boric acid is 2.5:1, stir and mix thoroughly and let stand for 12 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing boric acid.

[0548] 20 g of polyvinyl alcohol was dissolved in 1 L of water, and then 10 g of isomerase for isomerizing lactose into lactulose was added to the polyvinyl alcohol solution, and stirring was continued for 30 minutes. Then, the water solvent was evaporated at room temperature to obtain a polyvinyl alcohol film containing isomerase for isomerizing D-xylose into D-xylulose.

[0549] The polyvinyl alcohol component containing triborax and the polyvinyl alcohol film containing the epimerase for isomerizing lactose into lactulose are mixed according to a certain proportion to obtain a composition containing triborax and the epimerase for isomerizing lactose into lactulose.

[0550] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0551] In a 1 mL preparation system, 20 mM Tris-HCl buffer (pH 8.5), 1 M lactose, and 0.8 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 70°C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 24 hours, the production of lactulose was 850 mM, the production of epilactose was 13 mM, and the lactulose conversion rate was 85.0%.

[0552] Example 14 A composition catalyst containing boric acid, an epimerase for isomerizing lactose to lactulose, and polyvinyl alcohol Lactulose

[0553] A boric acid-containing composition comprises 60 parts by weight of polyvinyl alcohol, 10 parts by weight of boric acid, and 30 parts by weight of an isomerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0554] Dissolve polyvinyl alcohol in water to prepare a solution, then add a saturated boric acid solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to boric acid being 6:1, stir and mix thoroughly and let stand for 24 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing boric acid.

[0555] The polyvinyl alcohol component containing boric acid and the epimerase that isomerizes lactose into lactulose are mixed according to a certain ratio to obtain a composition containing boric acid and the epimerase that isomerizes lactose into lactulose.

[0556] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0557] In a 1 mL preparation system, 20 mM phosphate buffer (pH 7.0), 1 M lactose, and 1.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 80°C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted 48 hours, the production of lactulose was 865 mM, the production of epilactose was 10 mM, and the lactulose conversion rate was 86.5%.

[0558] Example 15 A combination of carboxyphenylboronic acid, an isomerase for isomerizing lactose into lactulose, and organosilicon Preparation of Lactulose from Lactose Catalyzed by Biocatalysis

[0559] A composition containing carboxyphenylboronic acid comprises 55 parts by weight of organosilicon, 15 parts by weight of carboxyphenylboronic acid, and 30 parts by weight of epimerase cells for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0560] 30 mg of cells containing the epimerase for isomerizing lactose into lactulose were resuspended in 20 mM phosphate buffer (pH 7.0), 54 mg of TEOS and 1 mg of 3-aminopropyltriethoxysilane (APTES) were added thereto, and 2% glutaraldehyde solution was added to the above solution after stirring for 2 hours, and stirring was continued for 2 hours. The cells were collected by centrifugation, washed with deionized water 3 times, and resuspended in 20 mM phosphate buffer (pH 7.0), 15 mg of carboxyphenylboronic acid was added, and the reaction was continued for 12 hours. After centrifugation and washing, a composition containing carboxyphenylboronic acid and the epimerase for isomerizing lactose into lactulose was obtained.

[0561] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0562] In a 1 mL preparation system, 20 mM phosphate buffer (pH 7.2), 2 M lactose, and 1.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 70°C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 36 hours, the production of lactulose was 1700 mM, the production of epilactose was 20 mM, and the lactulose conversion rate was 85.0%.

[0563] Example 16 A method of isomerizing lactose into lactulose containing magnesium metaborate, epimerase, chitosan and epoxy resin Lipid composition catalyzes lactose to produce lactulose

[0564] A composition containing magnesium metaborate comprises 20 parts by weight of chitosan, 60 parts by weight of epoxy resin, 17 parts by weight of magnesium borate and 3 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0565] 17 parts by weight of magnesium metaborate and 20 parts by weight of chitosan solution were mixed, and then the mixed solution was dripped into a 0.5 g / L sodium tripolyphosphate solution through a syringe to obtain a chitosan matrix containing magnesium metaborate.

[0566] Tris-HCl buffer (50mM pH 7.0) was added to 8g of epoxy resin, stirred for 30min, and then the resin was washed with deionized water to obtain an activated epoxy resin. The activated epoxy resin was then added to 1g / L of epimerase that catalyzes the isomerization of lactose to lactulose, placed on a shaker for 24h, and the enzyme was fixed on the epoxy resin. The protein content in the epoxy resin and enzyme supernatant was determined by the Bradford method, and the weight ratio of the enzyme to the epoxy resin was calculated by the subtraction method to be 1:20.

[0567] A chitosan matrix containing magnesium borate and an epoxy resin loaded with enzymes are mixed in proportion to obtain a composition containing magnesium metaborate, which contains 20 parts by weight of chitosan, 60 parts by weight of epoxy resin, 17 parts by weight of magnesium metaborate and 3 parts by weight of isomerase for isomerizing lactose into lactulose.

[0568] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0569] In a 1 mL preparation system, 50 mM phosphate buffer (pH 7.5), 0.5 M lactose, and 5.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 60°C. The production of lactulose and ipilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction reached 48 hours, the production of lactulose was 402 mM, the production of ipilactose was 5 mM, and the lactulose conversion rate was 80.4%.

[0570] Example 17 A method containing sodium borate, an isomerase for isomerizing lactose into lactulose, polyvinyl alcohol and chitosan Composition catalyzes lactose to produce lactulose

[0571] A sodium borate-containing composition comprises 8 parts by weight of sodium borate, 40 parts by weight of polyvinyl alcohol, 32 parts by weight of chitosan and 20 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0572] Dissolve polyvinyl alcohol in water to prepare a solution, then add a saturated sodium borate solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to sodium borate being 5:1, stir and mix thoroughly and let stand for 12 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing sodium borate.

[0573] Chitosan is dissolved in acetic acid solution to prepare a 1.6% chitosan solution, and the epimerase that isomerizes lactose into lactulose is dissolved in the chitosan solution so that the concentration of the epimerase that isomerizes lactose into lactulose is 1%. After ultrasonic degassing, the mixed solution is added dropwise to a 2% NaOH solution to form gel balls, which are repeatedly washed with deionized water to obtain a chitosan component containing the epimerase that isomerizes lactose into lactulose.

[0574] The polyvinyl alcohol component containing sodium borate and the chitosan component containing the epimerase that isomerizes lactose into lactulose are mixed to obtain a composition containing sodium borate and the epimerase that isomerizes lactose into lactulose.

[0575] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0576] In a 1 mL preparation system, 100 mM Tris-HCl buffer (pH 8.5), 0.5 M lactose, and 1.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 55 ° C. The production of lactulose and ipilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 12 hours, the production of lactulose was 425 mM, the production of ipilactose was 6 mM, and the lactulose conversion rate was 85%.

[0577] Example 18 A method of isomerizing lactose into lactulose containing carboxyphenylboronic acid, a diastereomerase, and a magnetic tetraoxide Preparation of lactulose from lactose catalyzed by a combination of tri-iron and silicon oxide

[0578] A composition containing carboxyphenylboronic acid comprises 5 parts by weight of carboxyphenylboronic acid, 57 parts by weight of magnetic ferrosoferric oxide, 30 parts by weight of inorganic silicon oxide particles, and 8 parts by weight of an isomerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0579] Prepare 50mM Tris-HCl (pH 8.5) solution, take 100mL Tris-HCl solution, add 200mg dopamine solution to fully dissolve, then disperse 5.2g magnetic ferroferric oxide nanoparticles in the above solution, and stir mechanically for 12h. Collect ferroferric oxide by magnet, and collect ferroferric oxide after repeated washing with deionized water. Add 100mL anhydrous methanol to the ferroferric oxide nanoparticles modified with polydopamine, and then add 1g carboxyphenylboric acid and react for 24h. Finally, collect ferroferric oxide modified with carboxyphenylboric acid by magnet to obtain ferroferric oxide component containing carboxyphenylboric acid.

[0580] The diastereomerase for isomerizing lactose to lactulose was dissolved in 20 mM phosphate buffer (pH 7.0) to a final concentration of 1 g / L. 8 mL of the above enzyme solution was added with 2 mL of 1 g / L polyethyleneimine with a molecular weight of 1800, and then 298 mg of tetraethyl orthosilicate (TEOS), and the mixture was stirred for 20 min. After centrifugation and washing, the precipitate was collected to obtain a silicon oxide component containing the diastereomerase for isomerizing lactose to lactulose.

[0581] The ferrosoferric oxide component containing carboxyphenylboronic acid and the silicon oxide component containing the epimerase for isomerizing lactose into lactulose are mixed according to a certain ratio to obtain a composition containing carboxyphenylboronic acid and the epimerase for isomerizing lactose into lactulose.

[0582] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0583] In a 1 mL preparation system, 100 mM HEPES buffer (pH 7.5), 0.3 M lactose, and 5.0 g / L of the boron-containing composition prepared above were reacted at a constant temperature of 65 ° C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction reached 8 hours, the production of lactulose was 258 mM, the production of epilactose was 4 mM, and the lactulose conversion rate was 86%.

[0584] Example 19 A combination containing calcium metaborate, an isomerase for isomerizing lactose into lactulose, and calcium alginate Preparation of Lactulose from Lactose Catalyzed by Biocatalysis

[0585] A composition containing calcium metaborate comprises 60 parts by weight of calcium alginate, 20 parts by weight of calcium metaborate, and 20 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0586] 20 mg of calcium metaborate was dispersed in 4 mL of 10 g / L sodium alginate solution. After ultrasonic degassing, the above solution was added dropwise to 4 g / L calcium chloride solution. After incubation for 4 hours, the calcium alginate gel spheres were filtered and washed with deionized water 3 times to obtain calcium metaborate-containing calcium alginate gel spheres.

[0587] 20 mg of lactose isomerase for isomerizing lactulose was dispersed in 2 mL of 10 g / L sodium alginate solution. After ultrasonic degassing, the above solution was added dropwise to 4 g / L calcium chloride solution. After incubation for 4 hours, the calcium alginate gel spheres were filtered and washed with deionized water 3 times to obtain calcium alginate gel spheres containing lactose isomerase for isomerizing lactulose.

[0588] Calcium alginate gel beads containing calcium borate and calcium alginate gel beads of epimerase for isomerizing lactose into lactulose are mixed according to a certain proportion to obtain a composition containing calcium metaborate and epimerase for isomerizing lactose into lactulose.

[0589] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0590] In a 1mL preparation system, 20mM Tris-HCL buffer (pH 8.5), 1M lactose, and 0.5g / L of the boron-containing composition prepared above were reacted at a constant temperature of 55°C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 24h, the production of lactulose was 830mM, the production of epilactose was 10mM, and the lactulose conversion rate was 83%.

[0591] Example 20 A combination containing borax, an isomerase for isomerizing lactose into lactulose, and polyvinyl alcohol Preparation of Lactulose from Lactose Catalyzed by Biocatalysis

[0592] A composition containing triborax comprises 55 parts by weight of polyvinyl alcohol, 35 parts by weight of triborax, and 10 parts by weight of epimerase for isomerizing lactose into lactulose. The preparation method thereof is as follows:

[0593] Polyvinyl alcohol is dissolved in water to prepare a dissolving solution, and the diastereomerase that isomerizes lactose into lactulose is dissolved in the polyvinyl alcohol solution. Then, a borax aqueous solution is added to the mixed solution, and the mass ratio of the polyvinyl alcohol, borax and the diastereomerase that isomerizes lactose into lactulose is 55:35:10. After being fully stirred and mixed, the mixture is allowed to stand for 24 hours. The precipitate of the mixed solution is taken and freeze-dried using a freeze dryer to obtain a composition containing borax and the diastereomerase that isomerizes lactose into lactulose.

[0594] The boron-containing composition is used to catalyze lactose to prepare lactulose, and the method is as follows:

[0595] In a 1mL preparation system, 100mM Tris-HCl buffer (pH 8.0), 1M lactose, and 2.0g / L of the boron-containing composition prepared above were reacted at a constant temperature of 65°C. The production of lactulose and epilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection showed that when the reaction lasted for 48h, the production of lactulose was 850mM, the production of epilactose was 12mM, and the lactulose conversion rate was 85%.

[0596] Example 21 A method comprising magnesium metaborate, an epimerase for isomerizing D-fructose to D-psicose, chitosan and Preparation of D-psicose from D-fructose catalyzed by epoxy resin composition

[0597] A composition containing magnesium metaborate comprises 20 parts by weight of chitosan, 60 parts by weight of epoxy resin, 17 parts by weight of magnesium metaborate and 3 parts by weight of epimerase for isomerizing D-fructose into D-psicose. The preparation method thereof is as follows:

[0598] 17 parts by weight of magnesium borate and 20 parts by weight of chitosan solution were mixed, and then the mixed solution was dripped into a 0.5 g / L sodium tripolyphosphate solution through a syringe to obtain a chitosan matrix containing magnesium metaborate.

[0599] Tris-HCl buffer (50mM pH 7.0) was added to 8g of epoxy resin, stirred for 30min, and then the resin was washed with deionized water to obtain an activated epoxy resin. The activated epoxy resin was then added to 1g / L epimerase that catalyzes the isomerization of D-fructose to D-psicose, placed on a shaker for 24h, and the enzyme was fixed on the epoxy resin. The protein content in the epoxy resin and the enzyme supernatant was determined by the Bradford method, and the weight ratio of the enzyme to the epoxy resin was calculated by the subtraction method to be 1:20.

[0600] The chitosan matrix containing magnesium metaborate and the epoxy resin loaded with enzyme are mixed in proportion to obtain a composition containing magnesium metaborate and epimerase for isomerizing D-fructose into D-psicose.

[0601] The boron-containing composition was used to catalyze the preparation of D-psicose from D-fructose. Compared with Comparative Example 4, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-psicose to the substrate D-fructose in Example 21 increased by 34.2%.

[0602] Example 22 A method containing sodium borate, isomerase for isomerizing D-glucose to D-fructose, polyvinyl alcohol and chitosan Composition catalyzes the production of D-fructose from D-glucose

[0603] A sodium borate-containing composition comprises 8 parts by weight of sodium borate, 40 parts by weight of polyvinyl alcohol, 32 parts by weight of chitosan and 20 parts by weight of isomerase for isomerizing D-glucose into D-fructose. The preparation method thereof is as follows:

[0604] Dissolve polyvinyl alcohol in water to prepare a solution, then add a saturated sodium borate solution to the polyvinyl alcohol solution, the mass ratio of polyvinyl alcohol to sodium borate being 5:1, stir and mix thoroughly and let stand for 12 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a polyvinyl alcohol component containing sodium borate.

[0605] Chitosan is dissolved in acetic acid solution to prepare a 1.6% chitosan solution, and isomerase for isomerizing D-glucose into D-fructose is dissolved in the chitosan solution to make the concentration of isomerase for isomerizing D-glucose into D-fructose be 1%. After ultrasonic degassing, the mixed solution is added dropwise to a 2% NaOH solution to form gel balls, which are repeatedly washed with deionized water to obtain a chitosan component containing isomerase for isomerizing D-glucose into D-fructose.

[0606] The polyvinyl alcohol component containing sodium borate and the chitosan component containing isomerase for isomerizing D-glucose into D-fructose are mixed to obtain a composition containing sodium borate and isomerase for isomerizing D-glucose into D-fructose.

[0607] The boron-containing composition was used to catalyze the preparation of D-fructose from D-glucose. Compared with Comparative Example 1, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-fructose to the substrate D-glucose in Example 22 increased by 13.2%.

[0608] Example 23 A method containing calcium metaborate, an isomerase for isomerizing D-glucose into D-fructose, and a D-fructose isomerization enzyme Preparation of D-psicose from D-glucose catalyzed by a combination of epimerase of D-psicose and calcium alginate

[0609] A composition containing calcium metaborate comprises 60 parts by weight of calcium alginate, 20 parts by weight of calcium metaborate, 10 parts by weight of isomerase for isomerizing D-glucose into D-fructose, and 10 parts by weight of epimerase for isomerizing D-fructose into D-psicose. The preparation method thereof is as follows:

[0610] 20 mg of calcium metaborate was dispersed in 4 mL of 10 g / L sodium alginate solution. After ultrasonic degassing, the above solution was added dropwise to 4 g / L calcium chloride solution. After incubation for 4 hours, the calcium alginate gel spheres were filtered and washed with deionized water 3 times to obtain calcium metaborate-containing calcium alginate gel spheres.

[0611] 10 mg of isomerase for isomerizing D-glucose into D-fructose and 10 mg of epimerase for isomerizing D-fructose into D-psicose were dispersed in 2 mL of 10 g / L sodium alginate solution. After ultrasonic degassing, the above solution was added dropwise into 4 g / L calcium chloride solution. After incubation for 4 hours, the calcium alginate gel balls were filtered and washed with deionized water three times to obtain calcium alginate gel balls containing isomerase for isomerizing D-glucose into D-fructose and epimerase for isomerizing D-fructose into D-psicose.

[0612] Calcium alginate gel balls containing calcium metaborate and calcium alginate gel balls containing isomerase for isomerizing D-glucose into D-fructose and epimerase for isomerizing D-fructose into D-psicose are mixed in proportion to obtain a composition containing calcium metaborate, isomerase for isomerizing D-glucose into D-fructose, and epimerase for isomerizing D-fructose into D-psicose.

[0613] The boron-containing composition was used to catalyze the preparation of D-psicose from D-glucose. Compared with Comparative Example 5, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-psicose to the substrate D-glucose in Example 23 increased by 30.1%.

[0614] Example 24 A composition containing borax, isomerase for isomerizing D-xylose to D-xylulose, and polyvinyl alcohol Catalytic production of D-xylulose from D-xylose

[0615] A composition containing triborax comprises 65 parts by weight of polyvinyl alcohol, 25 parts by weight of triborax, and 10 parts by weight of isomerase for isomerizing D-xylose to D-xylulose. The preparation method thereof is as follows:

[0616] Dissolve polyvinyl alcohol in water to prepare a solution, dissolve the isomerase that isomerizes D-xylose to D-xylulose in the polyvinyl alcohol solution, and then add a borax aqueous solution to the mixed solution, the mass ratio of the polyvinyl alcohol, borax and the isomerase that isomerizes D-xylose to D-xylulose being 65:25:10, stir and mix thoroughly and let stand for 24 hours, take the precipitate of the mixed solution, and freeze-dry it with a freeze dryer to obtain a composition containing borax and the isomerase that isomerizes D-xylose to D-xylulose.

[0617] The boron-containing composition was used to catalyze the preparation of D-xylulose from D-xylose. Compared with Comparative Example 2, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product D-xylulose to the substrate D-xylose in Example 24 increased by 27.9%.

[0618] Example 25 A method comprising isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate, and an amino resin Combination of lipids catalyzes the production of fructose 6-phosphate from glucose 6-phosphate

[0619] A composition containing aminophenylboronic acid comprises 7 parts by weight of aminophenylboronic acid, 90 parts by weight of amino resin, and 3 parts by weight of isomerase for isomerizing glucose 6-phosphate to fructose 6-phosphate. The preparation method thereof is as follows:

[0620] 9.0 g of amino resin was dispersed in 100 mL of 20% glutaraldehyde solution, stirred at room temperature for 12 h, filtered to collect the resin, washed with deionized water 3 times, and then the glutaraldehyde-modified resin was collected. Then, the glutaraldehyde-modified resin was dispersed in a solution containing 0.9 g of 2-aminophenylboronic acid, incubated for 24 h, filtered to collect the resin, and washed with deionized water to obtain a resin component containing aminophenylboronic acid.

[0621] The amino resin component containing aminophenylboronic acid and the isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate are mixed according to a certain ratio to obtain a composition containing aminophenylboronic acid and the isomerase that isomerizes glucose 6-phosphate to fructose 6-phosphate.

[0622] The boron-containing composition was used to catalyze glucose 6-phosphate to prepare fructose 6-phosphate. Compared with Comparative Example 6, under the same enzyme dosage, the same reaction and detection conditions, the ratio of the product fructose 6-phosphate to the substrate glucose 6-phosphate in Example 25 increased by 25.6%.

[0623] Example 26 Large-Scale Preparation of Lactulose

[0624] The boron-containing composition was prepared according to the method of Example 12. The prepared boron-containing composition was used to catalyze lactose to produce lactulose, as follows:

[0625] In a 5L preparation system, 100mM HEPES buffer (pH 7.5), 1M lactose, 10.0g / L of the boron-containing composition prepared above were reacted at 60°C. The production of lactulose and ipilactose was detected by high performance liquid chromatography. The detection method is shown in Example 11. HPLC detection shows that when the reaction is to 8h, the production of lactulose is 870mM, the production of ipilactose is 16mM, and the lactulose conversion rate is 87.0%. After the reaction is completed, the boron-containing composition is removed by filtering. The residues of boron compounds in the reaction mixture are determined according to the method of national standard GB 5009.275-2016 (Determination of boric acid in food safety national standard food). According to the method of national standard, no boron-containing compound residues were detected. In addition, the residues of boron element in the reaction mixture were determined again by ICP-MS, and the test results showed that no boron element was detected.

[0626] The experimental results show that after the reaction is completed, the boron-containing composition can be easily removed from the reaction solution by a simple separation method (such as filtration) without the need to use special resins for removing boron compounds, which is beneficial for simplifying the production process and reducing production costs; and there is no residual boron compound in the reaction solution.

[0627] It should be noted that, although the technical solution of the present invention is introduced with specific examples, those skilled in the art will appreciate that the present invention should not be limited thereto.

[0628] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or technical improvements in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A boron-containing composition comprising 50-90 parts by weight of a substrate, 5-35 parts by weight of a boric acid compound and / or a derivative thereof and 1-30 parts by weight of an enzyme.

2. The boron-containing composition according to claim 1, in, The boron-containing composition acts as a catalyst; Preferably, the boron-containing composition is used as a catalyst for the production of sugars and / or their derivatives.

3. The boron-containing composition according to claim 2, in, The sugar includes one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides; Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides; More preferably, the sugar includes one or more of monosaccharides and disaccharides.

4. The boron-containing composition according to claim 2 or 3, in, The sugar derivatives include one or more of phosphate sugars and deoxy sugars.

5. The boron-containing composition according to any one of claims 1 to 4, in, The matrix includes any substance capable of loading a boric acid compound and / or its derivatives and / or an enzyme. Preferably, the matrix includes one or more of an organic synthetic polymer, an inorganic substance, a natural organic macromolecular compound, a magnetic particle, a metal organic framework material, a covalent organic framework material, an organosiloxane polymer layer and a macroporous resin.

6. The boron-containing composition according to any one of claims 1 to 5, in, The boronic acid compound and / or its derivative is loaded on the substrate; Optionally, the enzyme is supported on the matrix.

7. The boron-containing composition according to any one of claims 1 to 6, in, The boronic acid compound and / or its derivative and the enzyme are respectively loaded on the same substrate; and / or, The boronic acid compound and / or its derivative and the enzyme are respectively loaded on different substrates; and / or, The boronic acid compound and / or its derivative and the enzyme are simultaneously loaded on the same substrate; and / or, The boric acid compound and / or its derivative is loaded on the matrix, and the enzyme is in a free form; Optionally, the loading is achieved by adsorption, embedding, cross-linking, covalent binding or a combination thereof.

8. The boron-containing composition according to any one of claims 1 to 7, in, The boron-containing composition further comprises a linker to load the boronic acid compound and / or its derivative and / or the enzyme on the substrate; Optionally, the linker is a cross-linker.

9. The boron-containing composition according to any one of claims 1 to 8, in, The enzyme comprises one or more enzymes from the group of isomerases belonging to EC5 in EC number; Optionally, the enzyme comprises one or more enzymes from the group of glycosyltransferases belonging to EC numbering EC 2.

4.

10. The boron-containing composition according to any one of claims 1 to 9, in, The enzymes include one, two, three or more enzymes having the same or different functions.

11. The boron-containing composition according to any one of claims 1 to 10, in, The boric acid compound and / or its derivatives include one or more of boric acid, inorganic borates, organic boric acid and organic borates.

12. The boron-containing composition according to any one of claims 1 to 11, in, The boron-containing composition may be in the form of one or more of beads, spheres, particles, granules, gels, fibers, films, or surfaces.

13. The method for preparing a boron-containing composition according to any one of claims 1 to 12, comprising the steps of: mixing a boric acid compound and / or its derivative and / or an enzyme with a substrate, and loading the substrate to obtain the boron-containing composition.

14. Any of the following uses of the boron-containing composition according to any one of claims 1 to 12 or the boron-containing composition prepared by the preparation method according to claim 13: (a) Use in the preparation of sugars and / or their derivatives; (b) Use as a catalyst for preparing sugars and / or their derivatives; Optionally, the sugar comprises one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides; the sugar derivative comprises one or more of phosphate sugars and deoxy sugars; Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides; More preferably, the sugar includes one or more of monosaccharides and disaccharides.

15. A method for preparing sugar and / or its derivatives, comprising the steps of: (i) preparing a reaction mixture comprising a substrate, (ii) contacting the reaction mixture with a boron-containing composition under suitable reaction conditions; in, The boron-containing composition is the boron-containing composition according to any one of claims 1 to 12 or the boron-containing composition prepared by the preparation method according to claim 13.

16. The method according to claim 15, in, The substrate includes sugar and / or its derivatives; Optionally, the sugar comprises one or more of monosaccharides, disaccharides, oligosaccharides and polysaccharides; the sugar derivative comprises one or more of phosphate sugars and deoxy sugars; Preferably, the sugar comprises one or more of monosaccharides, disaccharides and oligosaccharides; More preferably, the sugar includes one or more of monosaccharides and disaccharides.

17. The use or method according to any one of claims 14 to 16, in, Using a boron-containing composition to prepare sugar and / or its derivatives under acidic conditions, neutral conditions or alkaline conditions, Optionally, the acidic condition is a pH value lower than 7.0, the neutral condition is a pH value equal to 7.0, and the alkaline condition is a pH value higher than 7.

0.

18. The use or method according to any one of claims 14 to 17, in, In solution, using a boron-containing composition to prepare sugars and / or derivatives thereof, Optionally, the solution is selected from an aqueous solution, an organic solvent, an ionic liquid or a deep eutectic solvent, and / or, The solution is selected from a buffer or a non-buffer; Preferably, the buffer is selected from acetate buffer, citrate buffer, glycine buffer, phthalate buffer, phosphate buffer, 4-hydroxyethylpiperazineethanesulfonic acid buffer, 3-morpholinepropanesulfonic acid buffer, tris buffer, carbonate buffer or ammonia-ammonium chloride buffer.

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