107 results about "Pantoic acid" patented technology

The invention relates to a method used microorganism enzyme resolution DL-pantoic acid lactone to produce D-pantoic acid. It uses the D-pantoic acid lactone hydrolase strain of the fusarium, gibberella, aspergillus, penicillium, rhizopus, gliocladium, aureobasidium to ferment and culture, uses wet thallus as coarse enzyme, DL-pantoic acid lactone as substrate to produce D-pantoic acid. L- pantoic acid lactone can be reclaimed. The DL-pantoic acid lactone gained by racemization reaction can newly be used to do resolution.

The invention provides a method for preparing D-pantoyl lactone by fermentation. The method comprises the following steps: culturing fusarium oxysporum in a culture medium; (2) mixing the cultured fusarium oxysporum with a substrate and hydrolyzing to obtain D-pantoic acid; (3) carrying out lactonization on the D-pantoic acid in the step (2) to obtain the D-pantoyl lactone, wherein the culture medium is prepared from 1 percent to 10 percent of glycerol, 0.2 percent to 2 percent of peptone, 0.2 percent to 2 percent of yeast extract, 0.5 percent to 5 percent of maize slurry, 0.05 percent to 0.2 percent of spirulina polysaccharide, 0.1 percent to 1 percent of glycine and the balance of water.

The invention relates to a recovery method for D-calcium pantothenate mother liquor. The recovery method comprises the following steps: subjecting D-calcium pantothenate mother liquor to recovery of methanol; heating a product obtained after recovery of methanol, adjusting a pH value to 1-3, carrying out sufficient stirring, then performing cooling to room temperature, adjusting a pH value to 4-6,carrying out filtering, and respectively recovering precipitated calcium sulfate and a filtrate; adding ethyl acetate into the obtained filtrate, carrying out sufficient extracting, respectively collecting an ethyl acetate phase and a water phase, and recovering D-pantoic acid lactone in the ethyl acetate phase; and concentrating, cooling and crystallizing the obtained water phase, carrying out filtering and respectively recovering crystallized beta-aminopropionic acid and a filtrate containing D-calcium pantothenate. According to the method provided by the invention, the residual target products and a small amount of unreacted raw materials and auxiliary materials in the D-calcium pantothenate mother liquor are efficiently and fully recovered according to a specific sequence, so resourcewaste and environmental pollution are avoided; and the method has the advantages of easy availability of raw materials, environment-friendly and safe conditions, short reaction time, recyclable solvents, simple steps, high purity of recovered products and the like.

The invention belongs to the technical field of microbial fermentation and enzymatic conversion, particularly relates to a method for high-density culture of D-pantolactone hydrolase producing fungi,and further discloses a method for preparing D-pantoic acid by enzymatic conversion carried out with D-pantolactone hydrolase producing fungi. The method for high-density culture of D-pantolactone hydrolase producing fungi utilizes only conventional fusarium moniliforme in the prior art as an enzyme producing strain, effectively increases the biomass of the enzyme producing strain by optimizing aseed medium and a fermentation medium, and also effectively increases the enzyme of a crude enzyme solution and improves the relative enzyme activity of fungi quantity per unit.

The invention relates to the field of chemical synthesis and discloses a method for preparing D-calcium pantothenate. The method comprises the following steps of: reacting 3-aminopropionitrile with aqueous solution of sodium hydroxide or potassium hydroxide at the temperature of between 96 and 100 DEG C, adding hydrochloric acid to regulate the pH value, and concentrating and crystallizing; dissolving the prepared crystal by using methanol, and filtering to remove salt; and performing ion exchange with 732-type calcium ion exchange resin, carrying out acylation reaction between reaction liquid and D-pantolactone, crystallizing at the temperature of between 15 DEG C below zero and 10 DEG C below zero, and drying to prepare the D-calcium pantothenate, wherein the molar ratio of the 3-aminopropionitrile to the sodium hydroxide and the potassium hydroxide is 1:1.1-1.4, and the molar ratio of the 3-aminopropionitrile to the D-pantolactone 1:1-1.1. The method for preparing the D-calcium pantothenate has the advantages of not using calcium oxide or calcium hydroxide as a calcification agent, avoiding water generation in the reaction process so as to ensure the smooth acylation process, along with simple process, high yield, recycled 732-type calcium ion exchange resin, no generation of three wastes, and contribution to industrial production.

The invention relates to laevorotation lactone specificity hydrolytic enzyme producing strain and the method used it to make chirality oxyacid. The enzyme producing is Fusarium proliferatum Nirenberg ECU2002 with storage number CGMCC 1494. The chirality oxyacid preparing method includes the following steps: using the fungous mycelium, rough enzyme extract or their immobilization derivative as biocatalyst; processing antipode selectivity hydrolysis resolution for a series of racemic chirality lactone to gain many optically active (+)-oxyacid and (+)-lactone which can be hydrolyzed into (-)-oxyacid; (+)-alpha-hydroxyl-beta, beta-dimethyl-gamma-butyric acid which are D-(+)-pantoic acid; simple acidizing to gain chirality intermediate D-(-)-pantoic acid lactone widely used in preparing feed and daily chemical engineering industry.

The invention relates to a preparation method of a vitamin B5 crude product. The preparation method comprises the process steps of carrying out adsorption separation on DL-pantolactone enzymatic hydrolysate by virtue of macroporous adsorption resin, and carrying out purification, filtration, lactonization, solvent extraction, D-calcium pantothenate synthesis and vacuum drying, so as to obtain theVB5 crude product. The method for separating and extracting D-pantoic acid and L-pantolactone by virtue of the macroporous adsorption resin has beneficial effects that the problems that the quality and yield are reduced due to the emulsion easily occurring in an extraction process are overcome; meanwhile, VB5 is separated out by virtue of a solvent-method crystallization technique, so that the crystallization time is shortened, the extraction yield is increased, the product quality is improved, and the economic benefit is increased; and the method is simple and convenient, low in solvent loss,strong in operability and suitable for industrial production, and the environmental pollution can be reduced.

The invention relates to an immobilization method of D-pantoic acid lactone hydrolase, comprising the steps: (1) mycelium is pretreated to obtain the target enzyme liquid; (2) macroporous ion exchange resin is pretreated and added with glutaric dialdehyde solution for cross linking; (3) after that, the obtained macroporous ion exchange resin is suspended the D-pantoic acid lactone hydrolase solution and washed by de-ionized water to remove resolvase, so that the immobilized D-pantoic acid lactone hydrolase is obtained. The invention takes the macroporous ion exchange resin D201GF as a carrier and immobilizes the D-pantoic acid lactone hydrolase by two-step reaction of glutaric dialdehyde, thus overcoming the defect of the traditional direct reaction of mycelium. The immobilized enzyme granules have uniform shapes, good mechanical strength and large superficial area, so as to greatly improve the load capacity of enzyme and enhance enzyme activity; the recovery rate of enzyme activity reaches 85% in average, and the recycling times of the immobilized enzyme reaches above 50, so that the reaction cost of the resolvase is reduced. Furthermore, the method has simple technique and little investment for production equipment.

The invention provides a pantolactone hydrolase mutant strain and application thereof. Specifically, the invention provides a mutant pantolactone hydrolase with significantly improved enzyme activity and a host cell for expressing the mutant pantolactone hydrolase. The invention further provides an enzyme preparation containing the mutant pantolactone hydrolase, D-pantolactone in a substrate DL-pantolactone can be completely hydrolyzed into D-pantoic acid, and the mutant pantolactone hydrolase is high in conversion rate, stable in property, capable of being repeatedly used and suitable for industrial production.

The invention relates to the technical field of genetic engineering, in particular to L-panolactone dehydrogenase and application thereof. The L-panolactone dehydrogenase derived from Nocardia asteroides is disclosed for the first time, and the L-panolactone dehydrogenase has good enzymatic activity to L-panolactone, and can be applied to catalytic synthesis of D-panolactone. A system of multi-enzyme cascade catalysis for chiral overturning of the L-panolactone to synthesize the D-panolactone is established, genetically engineered bacterium cells for co-expression of the L-panolactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase are taken as a catalyst, accumulation and spontaneous hydrolysis of an intermediate product, namely ketopantolactone are avoided, the steps of separation of the intermediate product, racemization of the L-panolactone, lactonization of D-pantoic acid under the acid condition and the like are omitted, the reaction process is simplified, using of acid and base is reduced, and the reaction efficiency is improved.

The invention relates to a refining method of D-pantolactone. The method comprises the following steps: mixing a D-pantolactone crude product with the specific rotation of more than -45.0 degrees withethyl acetate, heating and fully dissolving, controlling the temperature at 20-30 DEG C, adding a D-pantolactone seed crystal with the specific rotation of -45.0 degrees to -50.7 degrees, slowly cooling to 5-10 DEG C, carrying out heat preservation crystallization, filtering, and collecting the crystal. The refining method of the D-pantolactone has the advantages that the raw materials are easy to obtain, the conditions are environmentally friendly and safe, the reaction time is short, the solvent can be recycled, the steps are simple, the obtained product is large and uniform in crystal, thespecific rotation can reach -49.5 degrees to -50.5 degrees, the purity can reach 98.0%, and the refining method has extremely high industrial utilization value.

The invention relates to the technical field of gene engineering, in particular to L-pantoyl lactone dehydrogenase and application of the L-pantoyl lactone dehydrogenase. The L-pantoyl lactone dehydrogenase derived from Nocardia farcinica is disclosed for the first time, has good enzymatic activity to L-pantolactone and can be applied to catalytic synthesis of D-pantolactone. A system for multi-enzyme cascade catalysis of chiral overturning of the L-pantolactone to synthesize the D-pantolactone is constructed, genetically engineered bacterium cells for co-expression of the L-pantoyl lactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase are taken as a catalyst, accumulation and spontaneous hydrolysis of an intermediate product, namely ketopantolactone, are avoided, the steps such as separation of the intermediate product, racemization of the L-pantolactone and lactonization of D-pantoic acid under the acid condition are omitted, the reaction process is simplified, using of acid and alkali is reduced, and the reaction efficiency is improved.

The invention relates to the technical field of genetic engineering, in particular to L-pantolactone dehydrogenase and application thereof. The L-pantolactone dehydrogenase derived from Nocardia cyriacigeorgica is disclosed for the first time, the L-pantolactone dehydrogenase has good enzymatic activity for L-pantolactone and can be applied to the catalytic synthesis of D-pantolactone. A system for synthesizing D-pantolactone through multi-enzyme cascade catalyzed L-pantolactone chiral inversion is constructed, genetically engineered bacterial cells which co-express L-pantolactone dehydrogenase, D-keto-pantolactone reducase and glucose dehydrogenase are used as catalysts, the accumulation and spontaneous hydrolysis of the intermediate keto-pantolactone are avoided, the separation of intermediates, racemization of L-pantolactone and D-pantoic acid lactonization under acidic conditions are omitted, the reaction process is simplified, the use of acid-base is reduced, and the reaction efficiency is improved.

The invention discloses a lactone hydrolase mutant in the fields of genetic engineering technology and biocatalysis, and application of the lactone hydrolase mutant as a biocatalyst to resolution of D, L-pantolactone to prepare optically pure D-pantoic acid. The lactone hydrolase mutant is obtained by carrying out single mutation or combined mutation on 1-14 amino acid sites in an amino acid sequence SEQ NO: 2 by taking fusarium oxysporum lactone hydrolase as a parent enzyme. The hydrolysis activity of the lactone hydrolase mutant on D, L-pantolactone is improved by 2.3-128 times compared withthat of parent enzyme. The high-activity lactone hydrolase provided by the invention can be used for large-scale production of preparing optically pure D-pantoic acid by biologically splitting D, L-pantolactone.

The invention discloses a strain of agrobacterium radiobacter of specificity hydrolytic dextral lactone and the application of the agrobacterium radiobacter in preparing dextral lactone compound such as D-(-)-pantoic acid lactone and the like. The enzyme-producing bacterium is the agrobacterium radiobacter and the preservation number is CGMCC2371. Thalli of the bacterium and an immobilized derivative of the bacterium are taken as biocatalyst, dexpazufloxacin which are not needed in pantoic acid lactone racemic mixture is hydrolyzed into (-)-hydroxy acid directly, useful D-(-)-pantoic acid lactone is preserved, and the latter is widely used in producing feedstuff and D-calcium pantothenate and D-panthenol which are useful in the daily chemical industry.

The invention belongs to the technical field of enzyme engineering, and specifically relates to a method for efficiently extracting D-pantoic acid from an enzymatic hydrolysate. According to the method for efficiently extracting D-pantoic acid from the enzymatic hydrolysate, on the basis of the prior extraction method of D-pantoic acid, the extraction of D-pantoic acid from the enzymatic hydrolysate by using existing ethyl acetate as an extracting agent is more thorough through further optimization of the extracting agent. For the technical process of taking D-pantoic acid as a target extract,the interference of other substances in the water phase after the extraction is effectively eliminated, so that the subsequent refining treatment process is simpler.

The invention discloses a D-pantolactone hydrolase producing bacteria in the technical field of bioengineering, and relates to a Thielavia sp. Zmu20201 serving as a microorganism, and the preservationnumber of the strain is CCTCC: M2020062. Meanwhile, the invention further discloses the D-pantolactone hydrolase from the Thielavia sp. Zmu20201 strain and an engineering bacterium strain of the D-pantolactone hydrolase, wherein the amino acid sequence number of the D-pantolactone hydrolase is as shown in SEQ ID NO.2. In addition, the invention also discloses an application of the Thielavia sp. Zmu20201 fermentation thallus or the lactone hydrolase gene engineering strain fermentation thallus as a biocatalyst in resolution of D, L-pantolactone to prepare optically pure D-pantoic acid.

The invention provides a novel process for preparing a D-pantothenic acid calcium salt and relates to a novel process for preparing a D-pantothenic acid calcium salt. According to the basic scheme ofthe invention, the novel process comprises the following steps of: preparing a water solution with the content of 16-26% from DL-pantolactone, adding a biological enzyme which is 5-15% of mass of thewater solution, performing uniform mixing, and then slowly dropwise adding a calcium di-beta-alaninate water solution in which the content of calcium di-beta-alaninate is 20-30% in a molar amount of 0.49-0.51 fold that of D-pantolactone contained in the water solution at 23-30 DEG C; performing reaction all the time until complete hydrolysis of the D-pantolactone, filtering out a clear liquid andrecycling the biological enzyme in a filter cake; keeping a pH value of a reaction system between 6.0 and 7.3 during dropwise adding; concentrating the clear liquid in vacuum of -0.085MPa or above andat 115-120 DEG C after reaction is completed, performing cooling to 65 DEG C or below, adding methanol of which the weight is 0.5-0.8 fold that of D-pantothenic acid calcium di-beta-alaninate, performing mixing and cooling to 20-40 DEG C, performing filtering, and washing, pulping and rewashing the filter cake with the methanol to achieve purification; collecting filtrate of a few times, recovering the methanol and then forming the DL-pantolactone in a racemization manner to reuse; and drying the filter cake through two levels of temperature zones of 105-125 DEG C and 150-160 DEG C, and performing cooling and discharging to obtain the D-pantothenic acid calcium salt. According to the novel process, a precursor substance D-pantothenic acid calcium di-beta-alaninate of the D-pantothenic acid calcium salt is generated in one step during resolution reaction, so that the process of extracting and separating the D-pantolactone is avoided.

The invention relates to a new fusarium verticillioides bacterial strain, and further relates to a method for high yield of D-(-)-Pantolactone hydrolase. The method comprises the steps of performing fermentation and culturing on the fusarium verticillioides bacterial strain of which the preservation No. is CGMCC No.18129, and extracting the D-(-)-Pantolactone hydrolase from fermentation products.The specific bacterial strain is used for fermentation production, so that D-(-)-Pantolactone hydrolase fermentation liquid being high in enzyme activity can be obtained, and the D-(-)-Pantolactone hydrolase being high in quality can be obtained efficiently.