Long chain compositions, combinations of long chain compositions, and methods of manufacture and applications thereof

Abstract

Disclosed are a long-chain composition or a combination of long-chain compositions, a method for producing the same, and its use in the production of long-chain dibasic acids by fermentation. The long-chain composition comprises one or more long-chain alkanes and one or more long-chain carboxylic acids. The one or more long-chain alkanes are selected from the group consisting of C9-18 linear or branched alkanes. The one or more long-chain carboxylic acids are selected from the group consisting of C9-18 linear or branched, saturated or unsaturated aliphatic monocarboxylic acids. The mass ratio of the long-chain alkane to the long-chain carboxylic acid is (1:1) to (40:1). Advantages of this long-chain composition include high fermentation efficiency or substrate utilization when used as a starting material for the production of long-chain dibasic acids by fermentation.

Description

[Technical field]

[0001] This application belongs to the field of biochemical engineering. In particular, it relates to a long chain composition or a combination of long chain compositions. This application also relates to a method for producing the long chain composition or the combination of long chain compositions and its application in the production of long chain dibasic acids. [Background technology]

[0002] Dodecanedioic acid (DDDA) is an aliphatic dicarboxylic acid with a carbon chain containing 12 carbon atoms. DDDA is an important and widely used fine chemical product in industry. In addition, DDDA is also an important starting material in the synthesis of high-quality fragrances, high-performance nylon engineering plastics, high-quality nylon hot melt adhesives, high-temperature dielectrics, high-quality paints and coatings, high-quality lubricants, cold-resistant plasticizers, resins, medicines, pesticides, etc. in the chemical industry.

[0003] In the method for producing long-chain dibasic acids by fermentation, the two terminal methyl groups of long-chain n-alkanes are converted to carboxyl groups by α- and ω-oxidation. This oxidation is carried out at room temperature and pressure using the specific oxidizing ability and oxidizing action of intracellular enzymes in microorganisms. As a result, various long-chain dibasic acids with corresponding chain lengths are synthesized. A wide variety of bacteria, fungi, and actinomycetes can produce long-chain dibasic acids by fermentation. Among them, yeasts of the genus Candida are highly productive microorganisms that ferment n-alkanes to produce dibasic acids.

[0004] In the prior art, common yeast is used as fermentation strain, and n-alkane is used as substrate to produce long-chain dibasic acid with corresponding carbon number.For example, CN102115767A, CN102115768A, etc. disclose the production scheme of undecadiacid and hexadecanedioic acid, respectively.In addition, CN103805643A discloses the production method of long-chain dibasic acid, which includes the steps of growing and culturing dibasic acid strain to obtain seed culture, growing and fermenting the seed culture, adding emulsified alkane during the fermentation process, removing microbial protein after the reaction is completed, and crystallizing the obtained long-chain dibasic acid product. Summary of the Invention

[0005] The inventors of the present application have found that the prior art still has a problem, which is that when producing long-chain dibasic acids using n-alkanes as substrates, the fermentation degree and substrate utilization rate are low. The present application has been completed based on this finding.

[0006] Specifically, the present application is directed to the following aspects of the solution: [Aspect 1] A long chain composition (preferably for use in fermentation) comprising one or more long chain alkanes and one or more long chain carboxylic acids, the one or more long chain alkanes are selected from the group consisting of C9-18, linear or branched (preferably linear) alkanes; Preferably, it is one or more long-chain alkanes selected from the group consisting of n-dodecane, n-tetradecane and n-hexadecane, in particular n-dodecane; the one or more long chain carboxylic acids are selected from the group consisting of aliphatic monocarboxylic acids that are C9-18, linear or branched (preferably linear), and saturated or unsaturated (preferably saturated); Preferably, one or more long-chain carboxylic acids selected from the group consisting of lauric acid, myristic acid and palmitic acid, in particular lauric acid; the mass ratio of the long-chain alkane to the long-chain carboxylic acid is from (1:1) to (40:1) (preferably from (2:1) to (20:1) or from (5:1) to (10:1)); Long chain composition. [Aspect 2] It also contains water, The mass of the water is 0.5 to 10 times (preferably 1 to 5 times or 1 to 3 times) the total mass of the long-chain alkane and the long-chain carboxylic acid. 2. A long chain composition according to any of the above or below embodiments. [Aspect 3] A long chain composition according to any one of the above or below embodiments, which satisfies one or more of the following (i) to (iii): (i) the pH of the composition is 5 to 12 (preferably, 7 to 10, 7.5 to 9, or 7.5 to 8.0); (ii) exists in the form of a liquid or a solid-liquid mixture at 32° C. (preferably exists in the form of a liquid, in particular an aqueous liquid); (iii) The number of carbon atoms in the long-chain alkane is the same as the number of carbon atoms in the long-chain carboxylic acid. [Aspect 4] A combination of long chain compositions, wherein n long chain compositions according to any of the above or below embodiments are present independently of each other (e.g., packaged or compartmentalized independently of each other), The above n is a positive integer of 2 to 40 (preferably, 4 to 20 or 5 to 10), The mass ratio of the long-chain alkane to the long-chain carboxylic acid in the i-th long-chain composition is R i (i is any positive integer from 2 to n), and the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the first long-chain composition is R 1 and the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the n-th long-chain composition is R n Then, R i-1 / R i ≧1 (preferably, R i-1 / R iThe values ​​are 1 to 20, 1.0001 to 10, 1.001 to 10, 1.01 to 10, 1.1 to 5 or 1.5 to 2), R 1 / R n >1 (preferably, R 1 / R n The values ​​of are 1.0001 to 30, 1.001 to 20, 1.01 to 10, 1.1 to 5 or 1.5 to 2), Combination of long chain compositions. [Aspect 5] A combination of long chain compositions according to any of the above or below embodiments, which satisfies one or more of the following (i) to (iii): (i) the n long chain compositions are prepared separately; (ii) the weight ratios of any two of the n long chain compositions, calculated based on the amount of long chain composition, are the same or different, preferably the same or substantially the same; (iii) the weight ratio of any two of the n long chain compositions, calculated based on the amount of long chain alkane, is the same or substantially the same. [Aspect 6] 1. A method for producing a long chain composition (preferably for use in fermentation) comprising one or more long chain alkanes and one or more long chain carboxylic acids, the method comprising: the one or more long chain alkanes are selected from the group consisting of C9-18, linear or branched (preferably linear) alkanes; Preferably, it is one or more long-chain alkanes selected from the group consisting of n-dodecane, n-tetradecane and n-hexadecane, in particular n-dodecane; the one or more long chain carboxylic acids are selected from the group consisting of aliphatic monocarboxylic acids that are C9-18, linear or branched (preferably linear), and saturated or unsaturated (preferably saturated); Preferably, one or more long-chain carboxylic acids selected from the group consisting of lauric acid, myristic acid and palmitic acid, in particular lauric acid; the mass ratio of the long-chain alkane to the long-chain carboxylic acid is from (1:1) to (40:1) (preferably from (2:1) to (20:1) or from (5:1) to (10:1)); The above-mentioned production method includes at least a mixing step of mixing the above-mentioned long-chain alkane and the above-mentioned long-chain carboxylic acid in the above-mentioned mass ratio (preferably, mixing until homogeneous) to obtain the above-mentioned long-chain composition, Optionally, the mixing step is carried out while heating (preferably, heating to 45 to 70° C. or 50 to 60° C.) and stirring (preferably, stirring at 50 to 250 rpm or 150 to 250 rpm). Manufacturing method. [Aspect 7] The following (i) and / or (ii) are met: (i) In the above mixing step, water and a pH adjuster (such as an alkali, in particular one or more selected from sodium hydroxide and potassium hydroxide) are further added; (ii) adding water and a pH adjuster (such as an alkali, in particular one or more selected from sodium hydroxide and potassium hydroxide) to the long-chain composition, optionally with heating (preferably to 45-70° C. or 50-60° C.) and stirring (preferably at 50-250 rpm or 150-250 rpm); the mass of the water is 0.5 to 10 times (preferably, 1 to 5 times or 1 to 3 times) the total mass of the long-chain alkane and the long-chain carboxylic acid; The amount of the pH adjuster used is an amount that adjusts the pH of the long chain composition to 5 to 12 (preferably 7 to 10, 7.5 to 9, or 7.5 to 8.0). A manufacturing method according to any one of the above or below embodiments. [Aspect 8] A method for producing a long-chain dibasic acid, comprising the steps of: The long-chain dibasic acid is one or more long-chain dibasic acids selected from the group consisting of C9 to C18, linear or branched (preferably linear), saturated or unsaturated (preferably saturated) aliphatic dicarboxylic acids; Preferably, it is one or more long-chain dibasic acids selected from the group consisting of dodecanedioic acid, tetradecanedioic acid and hexadecanedioic acid, in particular dodecanedioic acid; The above-mentioned production method includes the following steps (1) and (2): (1) producing a long chain composition by a method for producing the long chain composition described in any of the above or below embodiments to provide a long chain composition or a combination of long chain compositions described in any of the above or below embodiments (collectively referred to as a long chain composition); (2) fermenting the long chain composition in the presence of a fermentation bacterium and a fermentation medium to convert the long chain composition to long chain dibasic acids. [Aspect 9] The method for producing a composition according to any one of the above or below embodiments, which satisfies the following (i) and / or (ii): (i) the fermenting bacterium is a yeast having a complete α,ω oxidation pathway; Preferably, the yeast is one or more species selected from the group consisting of the genera Candida, Cryptococcus, Endomyces, Hansenula, Pichia, Rhodotorula, Torulopsis and Trichosporon, More preferably, it is one or more yeasts selected from the genus Candida. In particular, Candida tropicalis More particularly, the mutant strain PF-UV-56 of Candida tropicalis (deposited at China General Microbiological Culture Collection Center, accession number: CGMCC No. 0356); (ii) The fermentation medium comprises: 20-28 g / L sucrose; 0.8-1.5g / L corn steep liquor; 2.0-4.0 g / L yeast extract; Sodium chloride at 0.8-1.2 g / L; potassium dihydrogen phosphate at 3.0-3.5 g / L; magnesium sulfate at 1.2-1.8 g / L; 1.2-4.8 g / L urea; 1.5-2 g / L ammonium sulfate; and Sodium acetate at 1.5-1.8 g / L. [Aspect 10] The fermentation method according to any one of the above or below embodiments, comprising the steps (i) and (ii) of: (i) a preliminary step of mixing the fermentation bacteria and the fermentation medium and fermenting for 5 to 60 hours (preferably 10 to 40 hours, more preferably 20 to 30 hours or 24 hours) to obtain a fermentation raw material solution; (ii) a conversion step in which the long-chain composition is added to the fermentation raw material solution and fermented for 60 to 400 hours (preferably 100 to 300 hours, more preferably 100 to 160 hours or 100 to 140 hours) to convert the long-chain composition into the long-chain dibasic acid. [Aspect 11] The method for producing a composition according to any one of the above or below embodiments, which satisfies one or more of the following (i) to (vii): (i) adding a seed culture liquid of the fermentation bacteria at 2 to 30% by volume (preferably 5 to 20% by volume or 10 to 15% by volume) of the total liquid retention volume; (ii) the reaction temperature of the fermentation reaction is 25 to 37°C (preferably, 28 to 32°C); (iii) the fermentation reaction is carried out with stirring at 100 to 1000 rpm (preferably, 120 to 500 rpm or 150 to 300 rpm); (iv) the aeration rate of the fermentation reaction is 0.2 to 10.0 VVM (preferably, 0.2 to 2.0 VVM or 0.5 to 1.0 VVM); (v) the reaction time of the fermentation reaction is 65 hours or more (preferably more than 120 hours, more preferably 130 to 400 hours, 130 to 300 hours, 130 to 200 hours, or 138 to 160 hours); (vi) the amount of the long chain composition added is 100 to 1000 g per 1 L of total liquid holding volume (preferably 100 to 500 g or 150 to 250 g per 1 L of total liquid holding volume); (vii) The amount of the fermentation medium added is 12 to 80 g per 1 L of total liquid retention volume (preferably 32 to 50 g, 35 to 45 g, or 37 to 42 g per 1 L of total liquid retention volume). [Aspect 12] The method for producing a composition according to any one of the above or below embodiments, which satisfies the following (i) and / or (ii): (i) during the preliminary step (preferably within 24 hours of beginning the preliminary step), the long chain composition is not added and the pH of the fermentation reaction is allowed to self-regulate (i.e., the pH is not adjusted); (ii) At the start of the conversion step (preferably 24 hours after the start of the preliminary step), the pH of the fermentation reaction is adjusted to 6.0 to 7.5 (preferably 6.8 to 7.0), and then the pH of the fermentation reaction is increased by 0.05 to 0.4 (preferably 0.1 to 0.3) at intervals of 5 to 60 hours (preferably 10 to 40 hours, 20 to 30 hours, or 24 hours) until the end of the fermentation reaction. [Aspect 13] The method for producing a composition according to any one of the above or below embodiments, which satisfies the following (i) or (ii): (i) adding said long chain composition in n batches in said converting step; The time intervals between two adjacent batches may be the same or different; Preferably, the time is 5 to 60 hours, 10 to 40 hours, 20 to 30 hours, or 24 hours. The above n is a positive integer of 2 to 40 (preferably, 4 to 20 or 5 to 10); (ii) In the conversion step, the long chain composition is added continuously depending on the reaction time. [Aspect 14] A method for producing a composition according to any one of the above or below embodiments, using a combination of long chain compositions according to any one of the above or below embodiments, which satisfies the following (i) or (ii): (i) adding n of the long chain compositions in n batches (preferably in the order of the first long chain composition through the nth long chain composition); (ii) adding said long chain composition continuously according to a function R=f(t); During the ceremony, R is the mass ratio of the long chain alkane to the long chain carboxylic acid; t is the reaction time of the conversion step; f() represents any non-increasing function (preferably any decreasing function, any monotonically decreasing function or any linearly decreasing function). [Aspect 15] The method according to any one of the above or below aspects, further comprising a step of separating the long-chain dibasic acid from a reaction product of the fermentation reaction.

[0007] The present application is also directed to the following aspects of the solution: <Aspect 1> A method for producing dodecanedioic acid, comprising the following steps (1) to (3): (1) mixing dodecane and lauric acid in a predetermined ratio, heating and stirring to completely dissolve the lauric acid, adding water to the mixture, mixing uniformly, and adding an alkali to adjust the pH to 7-9 to obtain a mixed substrate; (2) charging the fermentation bacteria and fermentation medium into a fermentation tank for fermentation, and adding the mixed substrate obtained in step (1) during the fermentation process until the fermentation is completed; (3) After the fermentation is completed, the reaction mixture is subjected to demulsification, membrane filtration, acid precipitation, filtration and drying to obtain the long-chain dibasic acid product. <Aspect 2> In the above step (1), The mass ratio of the dodecane to the lauric acid is from (2:1) to (10:1), and preferably from (5:1) to (10:1). A manufacturing method according to any one of the above or below aspects. <Aspect 3> In the above step (1), The heating temperature is 45 to 70°C, preferably 50 to 60°C. The stirring speed is 50 to 250 rpm, preferably 150 to 250 rpm. A manufacturing method according to any one of the above or below aspects. <Aspect 4> In the above step (1), Add water in an amount of 1 to 3 times the mass of the above mixture. A manufacturing method according to any one of the above or below aspects. <Aspect 5> In the above step (1), After uniform mixing, the above alkali is added to adjust the pH to 7 to 9, preferably 7.5 to 8.0. The alkali is one or more of sodium hydroxide and potassium hydroxide. A manufacturing method according to any one of the above or below aspects. <Aspect 6> The fermentation bacterium used in step (2) is a yeast having a complete α,ω oxidation pathway, Preferably, the bacterium is one or more of the genera Candida, Cryptococcus, Endomyces, Hansenula, Pichia, Rhodotorula, Torulopsis or Trichosporon. A manufacturing method according to any one of the above or below aspects. <Aspect 7> The method for producing the present invention according to any one of the above-mentioned or following embodiments, characterized in that the composition of the fermentation medium used in the step (2) is as follows: sucrose at 20-28 g / L; 0.8-1.5g / L corn steep liquor; 2.0-4.0 g / L yeast extract; Sodium chloride at 0.8-1.2 g / L; potassium dihydrogen phosphate at 3.0-3.5 g / L; magnesium sulfate at 1.2-1.8 g / L; 1.2-4.8 g / L urea; 1.5-2 g / L ammonium sulfate; and Sodium acetate at 1.5-1.8 g / L. <Aspect 8> In the above step (2), The amount of the seed culture liquid of the fermentation bacteria added is 5 to 20% of the volume of the fermentation liquid, and preferably 10 to 15%. A manufacturing method according to any one of the above or below aspects. <Aspect 9> In the above step (2), The fermentation temperature is 25 to 37°C, preferably 28 to 32°C. The stirring speed is 120 to 500 rpm, preferably 150 to 300 rpm. The ventilation rate is 0.2 to 1.0 VVM, preferably 0.5 to 1.0 VVM. The fermentation time is 138 to 144 hours. A manufacturing method according to any one of the above or below aspects. <Aspect 10> The method for producing the present invention according to any one of the above-mentioned or following embodiments, characterized in that in the step (2), the pH is adjusted as follows: 0-24 hours after fermentation: Do not adjust pH; After 24 hours: Adjust to a neutral pH range of 6.8-7.0; Thereafter: increase the pH by 0.1-0.3 at 24 hour intervals until the end of the reaction. <Aspect 11> In the above step (2), During the fermentation process, the mixed substrate obtained in step (1) is added in a pulsed or continuous feed mode. A manufacturing method according to any one of the above or below aspects. <Aspect 12> The method for producing a tertiary amine according to any one of the above-mentioned or later-mentioned embodiments, characterized in that in the step (2), the following (i) or (ii) is satisfied: Adopt intermittent feeding mode, complete feeding in 5-10 batches; The continuous feeding mode is adopted, and the feeding rate is calculated according to the fermentation time. <Aspect 13> In the above step (3), demulsification is carried out. The pH of the fermentation liquid is adjusted to 8.5 to 10, preferably 9 to 9.5. Heat the fermentation liquid to 75-90℃ and hold for 20-40 minutes. A manufacturing method according to any one of the above or below aspects. <Aspect 14> In the membrane filtration in the above step (3), The demulsified culture liquid is filtered through a membrane and separated into solid and liquid to obtain a clear fermentation liquid. The filtration temperature is 30 to 50°C, preferably 40 to 50°C. The pore size of the membrane is 10 to 50 nm, preferably 20 to 25 nm. A manufacturing method according to any one of the above or below aspects. <Aspect 15> In the acid precipitation in the above step (3), The pH of the clear fermentation liquid is adjusted with an acidic pH adjuster; The pH is adjusted to 3 to 5, preferably 3 to 4, to precipitate the long-chain dibasic acid. A manufacturing method according to any one of the above or below aspects. <Aspect 16> In the filtration in the above step (3), Filtration is carried out using a plate and frame filter. The filtration pressure is 0.5 to 1.0 MPa. The filtration temperature is 20 to 30°C. The drying temperature is 80 to 105°C. Drying time is 5 to 20 hours. A manufacturing method according to any one of the above or below aspects. Effect of the Invention

[0008] Compared with the prior art, the present application achieves one or more, and preferably all, of the following technical advantages: (1) In the long-chain composition of the present application, the mass ratio of long-chain alkane to long-chain carboxylic acid meets the requirements set forth in the present application. The use of the long-chain composition in a fermentation process can improve the fermentation degree. It can also solve the problem that long-chain carboxylic acid is poorly water-soluble and difficult to utilize in a fermentation system. (2) In the preferred embodiment of the long-chain composition of the present application, two steps, namely physical dissolution and chemical dissociation, are adopted, which can improve the dissolving effect of long-chain carboxylic acid, promote the formation of polar and soluble micelles by long-chain alkanes and long-chain carboxylic acids, and further improve the fermentation degree. (3) In the preferred embodiment of the method for producing the long-chain composition of the present application, an alkali adjustment scheme is adopted, which can reduce the pH fluctuation in the fermentation process caused by the supply of materials. In addition, in the continuous supply mode, the materials are further supplied, which can improve the substrate utilization rate and the degree of fermentation. (4) In the fermentation method of the present application, the long-chain carboxylic acids also function as carbon sources. In addition, some of the long-chain carboxylic acids can be directly converted into important intermediate metabolic products (acetyl coenzyme A) through fatty acid metabolism. This allows for rapid accumulation of biomass, shortens the fermentation time, and increases the production intensity of long-chain dicarboxylic acids. (5) In a preferred embodiment of the fermentation method of the present application, the fermentation degree and production intensity of long-chain dicarboxylic acids can be further improved by increasing the amount of long-chain fatty acids added during fermentation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present application will be described in detail below with reference to the embodiments. However, it should be noted that the scope of the present application is not limited to the following embodiments. Instead, the scope of the present application is defined by the appended claims.

[0010] All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.In the event of any conflict, the contents of this specification, including definitions, shall prevail.

[0011] When a material, method, component, instrument, or device described in this specification is modified by phrases such as "known to those of ordinary skill in the art" or "well known in the art," it is understood that the material, method, component, instrument, or device in question includes not only those previously used in the art at the time of the filing of this application, but also those that are not currently in common use but that later become known in the art and are suitable for similar purposes.

[0012] In the context of this application, the term "substantially" refers to an acceptable deviation or a deviation that would be considered reasonable by a person skilled in the art, for example, within ±10%, within ±5%, within ±1%, within ±0.5%, or within ±0.1% is acceptable to exist.

[0013] In the context of this application, the term "total liquid hold-up volume" means the total volume of the liquid phase in the fermentation vessel, which is usually 70-80% of the volume of the fermentation vessel.

[0014] In the context of this application, all percentages, parts, ratios, etc. are expressed on a weight basis, unless specifically stated to the contrary, and all pressures are expressed as gauge pressures.

[0015] In the context of this application, any two or more embodiments of this application may be combined. The technical solutions resulting from the combination also form part of the initial disclosure of this application and are included in the scope of this application.

[0016] According to an embodiment, the present application relates to a long chain composition. The long chain composition is preferably a long chain composition for use in fermentation. In particular, the long chain composition is suitable for use as a starting material in the production of long chain dibasic acids by fermentation.

[0017] According to an embodiment of the present application, the long-chain composition comprises one or more long-chain alkanes and one or more long-chain carboxylic acids. The one or more long-chain alkanes are selected from the group consisting of C9-18 linear or branched alkanes. The one or more long-chain carboxylic acids are selected from the group consisting of C9-18 linear or branched, saturated or unsaturated aliphatic monocarboxylic acids. The long-chain alkanes are preferably one or more long-chain alkanes selected from the group consisting of C9-18 linear alkanes. The long-chain alkanes are more preferably one or more long-chain alkanes selected from the group consisting of n-dodecane, n-tetradecane and n-hexadecane. The long-chain alkane is particularly preferably n-dodecane. The long-chain carboxylic acids are preferably one or more long-chain carboxylic acids selected from the group consisting of C9-18 linear saturated aliphatic monocarboxylic acids. The long-chain carboxylic acids are particularly preferably one or more long-chain carboxylic acids selected from the group consisting of lauric acid, myristic acid and palmitic acid. In particular, the long chain carboxylic acid is lauric acid.

[0018] According to an embodiment of the present application, the mass ratio of the long-chain alkane to the long-chain carboxylic acid is generally from (1:1) to (40:1), and preferably from (2:1) to (20:1) or from (5:1) to (10:1).

[0019] According to an embodiment of the present application, the long-chain composition further comprises water. The mass of water is generally 0.5 to 10 times, preferably 1 to 5 times or 1 to 3 times the total mass of the long-chain alkane and the long-chain carboxylic acid.

[0020] According to an embodiment of the present application, the pH of the long-chain composition is generally 5 to 12, preferably 7 to 10, 7.5 to 9 or 7.5 to 8.0. In the context of the present application, the method for measuring the pH of the long-chain composition may be a pH test paper method or a glass electrode method.

[0021] According to an embodiment of the present application, the long chain composition is present in the form of a liquid or a solid-liquid mixture at 32° C., preferably in the form of a liquid, in particular in the form of an aqueous liquid. In the context of the present application, the term "liquid" includes both homogeneous and non-homogeneous liquids, examples of the latter being multi-phase liquid-like mixtures of two or more liquids.

[0022] According to an embodiment of the present application, in the long chain composition, the long chain alkane and the long chain carboxylic acid have the same number of carbon atoms (such as a combination of n-dodecane and lauric acid).

[0023] According to an embodiment of the present application, the present application also relates to a combination of long chain compositions. The combination of long chain compositions includes a plurality of long chain compositions according to an embodiment of the present application. Specifically, the combination of long chain compositions includes n kinds of long chain compositions according to any embodiment of the present application, which are independent of each other. Here, n is generally a positive integer of 2 to 40, preferably a positive integer of 4 to 20 or 5 to 10. In addition, the expression "independently of each other" specifically means, for example, that the compositions are packaged independently of each other or are separated independently of each other (i.e., are divided into multiple fractions in the same package).

[0024] According to this embodiment, the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the i-th long-chain composition is R i and the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the first long-chain composition is R 1 The mass ratio of the long-chain alkane to the long-chain carboxylic acid in the n-th long-chain composition is R n Then, R i-1 / R i is generally 1 or more, and preferably 1 to 20, 1.0001 to 10, 1.001 to 10, 1.01 to 10, 1.1 to 5, or 1.5 to 2. Here, i is an arbitrary positive integer from 2 to n.

[0025] According to this embodiment of the present application, preferably, R 1 / Rn is greater than 1, and is preferably 1.0001 to 30, 1.001 to 20, 1.01 to 10, 1.1 to 5, or 1.5 to 2.

[0026] According to an embodiment of the present application, in the combination of long chain compositions, the n kinds of long chain compositions are each prepared separately. The preparation method of the long chain composition is described later in this specification.

[0027] According to an embodiment of the present application, in the combination of long chain compositions, the weight ratio (calculated based on the amount of long chain composition) of any two of the n long chain compositions may be the same or different, and preferably is the same or substantially the same.

[0028] According to other embodiments of the present application, the weight ratio (calculated based on the amount of long chain alkane) of any two of the n long chain compositions included in the combination of long chain compositions may be the same or substantially the same.

[0029] According to an embodiment of the present application, the long chain composition may be prepared by one method. For example, the preparation method includes at least the following mixing steps: Mixing step: A step of mixing the long-chain alkane and the long-chain carboxylic acid in the above-mentioned mass ratio to obtain a long-chain composition. In an optional configuration, mixing is performed while heating and stirring. Regarding heating, the heating temperature may be 45 to 70°C or 50 to 60°C. Regarding stirring, the stirring speed may be 50 to 250 rpm or 150 to 250 rpm. Furthermore, it is preferable to mix until the mixture becomes uniform, at which point a uniform liquid mixture is obtained.

[0030] According to an embodiment of the present application, water and a pH adjusting agent are further added in the mixing step of the preparation method, the pH adjusting agent being, for example, an alkali, in particular, one or more selected from sodium hydroxide and potassium hydroxide.

[0031] According to an embodiment of the present application, the preparation method further includes a step of adding water and a pH adjuster to the long chain composition. Optionally, this step is performed under heating and stirring. For heating, the heating temperature may be 45-70°C or 50-60°C. Furthermore, for stirring, the stirring speed may be 50-250 rpm or 150-250 rpm. The pH adjuster is, for example, an alkali, in particular one or more selected from sodium hydroxide and potassium hydroxide.

[0032] According to an embodiment of the present application, the mass of water to be added is generally 0.5 to 10 times, and preferably 1 to 5 times or 1 to 3 times, the total mass of the long-chain alkane and the long-chain carboxylic acid.

[0033] According to an embodiment of the present application, the amount of pH adjuster added is generally an amount that gives a pH of the long chain composition of 5 to 12, preferably an amount that gives a pH of 7 to 10, 7.5 to 9, or 7.5 to 8.0.

[0034] According to an embodiment of the present application, the present application also relates to a method for producing a long-chain dibasic acid. The long-chain dibasic acid is one or more long-chain dibasic acids selected from the group consisting of C9-18, linear or branched, saturated or unsaturated aliphatic dicarboxylic acids. The long-chain dibasic acid is preferably one or more long-chain dibasic acids selected from the group consisting of C9-18 linear saturated aliphatic dicarboxylic acids. The long-chain dibasic acid is preferably one or more long-chain dibasic acids selected from the group consisting of dodecanedioic acid, tetradecanedioic acid and hexadecanedioic acid. In particular, the long-chain dibasic acid is dodecanedioic acid.

[0035] According to an embodiment of the present application, the above-mentioned manufacturing method includes the following steps. (1) providing a long chain composition or a combination of long chain compositions (collectively referred to as long chain compositions) according to any one of the embodiments of the present application. (2) fermenting the long chain composition in the presence of a fermentation bacterium and a fermentation medium to convert the long chain composition to a long chain dibasic acid.

[0036] According to an embodiment of the present application, in the above-mentioned production method, the fermentation bacterium is a yeast having a complete α,ω oxidation pathway. Preferably, the fermentation bacterium is one or more yeasts selected from the group consisting of Candida, Cryptococcus, Endomyces, Hansenula, Pichia, Rhodotorula, Torulopsis and Trichosporon. More preferably, the fermentation bacterium is one or more yeasts selected from the genus Candida. In particular, the fermentation bacterium is Candida tropicalis. More particularly, the fermentation bacterium is a mutant strain PF-UV-56 of Candida tropicalis (deposited at China General Microbiological Culture Collection Center, deposit number: CGMCC No. 0356, August 31, 1998).

[0037] According to an embodiment of the present application, in the above-mentioned production method, the fermentation medium may be a conventional fermentation medium used in the present technical field in producing long-chain dibasic acids by fermentation. The fermentation medium generally contains 10-60 g / L of a carbon source, 1-10 g / L of a nitrogen source, 1-10 g / L of a phosphorus source, 0.1-50 ppm of a trace metal element source, etc. Examples of carbon sources include one or more selected from the group consisting of glucose, sucrose, maltose, fructose, molasses, glycerol, sorbitol, arabinose, rhamnose, methanol, and ethanol. Examples of nitrogen sources include one or more selected from the group consisting of yeast extract, peptone, corn steep liquor, urea, ammonium salts, and nitrates. Examples of phosphorus sources include one or more selected from the group consisting of orthophosphate, monohydrogen phosphate, and dihydrogen phosphate. Preferably, the phosphorus source is one or more selected from the group consisting of potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ammonium dihydrogen phosphate, diammonium hydrogen phosphate, sodium dihydrogen phosphate and disodium hydrogen phosphate. Examples of trace metal element sources include one or more selected from the group consisting of sulfates, hydrochlorides, acetates and nitrates of sodium, potassium, calcium, magnesium, iron, copper, zinc and manganese. The trace metal element source is preferably one or more selected from the group consisting of sodium chloride, sodium acetate, potassium chloride, magnesium sulfate, calcium chloride, iron chloride and copper sulfate. Particularly preferably, the fermentation medium contains the following components: 20-28 g / L sucrose, 0.8-1.5 g / L corn steep liquor, 2.0-4.0 g / L yeast extract, 0.8-1.2 g / L sodium chloride, 3.0-3.5 g / L potassium dihydrogen phosphate, 1.2-1.8 g / L magnesium sulfate, 1.2-4.8 g / L urea, 1.5-2 g / L ammonium sulfate and 1.5-1.8 g / L sodium acetate.

[0038] According to an embodiment of the present application, in the above-mentioned production method, the fermentation reaction includes at least the following two steps: Preliminary step: a step of mixing a fermentation bacterium and a fermentation medium and fermenting for 5 to 60 hours to obtain a fermentation raw material solution. The reaction time in the preliminary step is preferably 10 to 40 hours, more preferably 20 to 30 hours or 24 hours. Conversion step: a step of adding a long-chain composition to a fermentation raw material solution and fermenting for 60 to 400 hours to convert the long-chain composition into a long-chain dibasic acid. The reaction time of the conversion step is preferably 100 to 300 hours, more preferably 100 to 160 hours or 100 to 140 hours.

[0039] According to an embodiment of the present application, in the above-mentioned production method, the amount of the seed culture liquid of the fermentation bacteria added is generally 2 to 30 volume % relative to the total liquid retention volume, and preferably 5 to 20 volume % or 10 to 15 volume %.

[0040] According to an embodiment of the present application, in the above-mentioned production method, the reaction temperature of the fermentation reaction is generally 25 to 37°C, and preferably 28 to 32°C.

[0041] According to an embodiment of the present application, in the above-mentioned production method, the stirring speed of the fermentation reaction is generally 100 to 1000 rpm, and preferably 120 to 500 rpm or 150 to 300 rpm.

[0042] According to an embodiment of the present application, in the above-mentioned production method, the aeration rate in the fermentation reaction is generally 0.2 to 10.0 VVM, preferably 0.2 to 2.0 VVM or 0.5 to 1.0 VVM.

[0043] According to an embodiment of the present application, in the above-mentioned production method, the reaction time of the fermentation reaction is generally 65 hours or more, preferably more than 120 hours, more preferably 130 to 400 hours, 130 to 300 hours, 130 to 200 hours, or 138 to 160 hours, where the reaction time includes the reaction time of the preliminary step and the reaction time of the conversion step.

[0044] According to an embodiment of the present application, in the above production method, the amount of the long chain composition added is generally 100 to 1000 g, preferably 100 to 500 g or 150 to 250 g per 1 L of total liquid holding volume.

[0045] According to an embodiment of the present application, in the above-mentioned production method, the amount of fermentation medium input is usually 12-80 g, preferably 32-50 g, 35-45 g, or 37-42 g per liter of total liquid holding volume.

[0046] According to an embodiment of the present application, in the above-mentioned production method, no long chain composition is added in the preliminary step (preferably within 24 hours from the start of the preliminary step) and the pH of the fermentation reaction is allowed to self-regulate, i.e., no pH adjustment is performed during this step or period.

[0047] According to an embodiment of the present application, in the above-mentioned production method, at the start of the conversion step (preferably 24 hours after the start of the preliminary step), the pH of the fermentation reaction is adjusted to 6.0 to 7.5, preferably 6.8 to 7.0. More preferably, the pH of the fermentation reaction is then increased by 0.05 to 0.4, preferably 0.1 to 0.3. The time interval for increasing the pH is 5 to 60 hours, preferably 10 to 40 hours, 20 to 30 hours, or 24 hours, until the end of the fermentation reaction.

[0048] According to an embodiment of the present application, in the above-mentioned production method, in the conversion step, the long chain composition is added in n batches (also referred to as intermittent feeding). The time intervals between adjacent batches may be the same or different. Preferably, the time intervals are the same. Examples of the time intervals include 5 to 60 hours, 10 to 40 hours, 20 to 30 hours, and 24 hours. n is a positive integer of 2 to 40, preferably a positive integer of 4 to 20 or 5 to 10.

[0049] According to an embodiment of the present application, in the above-mentioned preparation method, in the conversion step, the combination of any one of the long chain compositions of the embodiments of the present application is used, and n kinds of long chain compositions are added in n batches, preferably, the n kinds of long chain compositions are added in the order from the first long chain composition to the nth long chain composition.

[0050] According to an embodiment of the present application, in the above-mentioned method, in the conversion step, the long chain composition is added continuously according to the reaction time (also called continuous feeding). The continuous feeding may be at a constant flow rate or not. Preferably, the continuous feeding is at a constant flow rate.

[0051] According to an embodiment of the present application, in the above-mentioned preparation method, in the conversion step, the long-chain composition is continuously added according to a function R=f(t), where R represents the mass ratio of the long-chain alkane to the long-chain carboxylic acid, and t represents the reaction time of the conversion step. f() represents any non-increasing function. Preferably, f() represents any decreasing function, any monotonically decreasing function, or any linearly decreasing function.

[0052] According to an embodiment of the present application, the above-mentioned preparation method further includes a step of separating the long-chain dibasic acid from the reaction product of the fermentation reaction (referred to as the fermentation liquor). For example, after the completion of the fermentation reaction, the resulting reaction product is subjected to demulsification, membrane filtration, acid precipitation, filtration and drying to obtain the long-chain dibasic acid product.

[0053] According to an embodiment of the present application, the demulsification may be performed by any conventional method known in the art, and the method is not particularly limited. For example, the demulsification may be performed by adjusting the pH of the fermentation liquid to 8.5 to 10 (preferably 9 to 9.5), increasing the temperature to 75 to 90°C, and maintaining it for 20 to 40 minutes.

[0054] According to an embodiment of the present application, membrane filtration may be performed by any conventional method known in the art, and the method is not particularly limited. For example, membrane filtration may be performed by demulsifying the fermentation liquid, filtering the fermentation liquid through a membrane, and then performing solid-liquid separation to obtain a clear fermentation liquid. The filtration temperature is generally 30 to 50°C, preferably 40 to 50°C. The pore size of the membrane is generally 10 to 50 nm, preferably 20 to 25 nm.

[0055] According to an embodiment of the present application, acid precipitation may be performed by any conventional method known in the art, and the method is not particularly limited. For example, acid precipitation may be performed by adjusting the pH of the clear fermentation liquor to 3-5 (preferably 3-4) using an acidic pH adjuster to precipitate the long-chain dibasic acid. The acidic pH adjuster is generally a strong acid, and may be one or more of sulfuric acid, hydrochloric acid, nitric acid, etc.

[0056] According to an embodiment of the present application, filtration may be performed by any conventional method known in the art, and the method is not particularly limited. For example, filtration may be performed by plate and frame filtration. The filtration pressure is generally 0.5 to 1.0 MPa. The filtration temperature is generally room temperature (e.g., 20 to 30°C).

[0057] According to the embodiment of the present application, drying may be performed by any conventional method known in the art, and the method is not particularly limited. For example, the drying temperature may be 80 to 105° C. The drying time may be 5 to 20 hours. EXAMPLES

[0058] The present application will be described in more detail below with reference to examples, but the present application is not limited to the following examples.

[0059] The experimental procedures employed in the following examples and comparative examples are conventional in the art unless otherwise noted. The experimental materials used in the following examples and comparative examples are commercially available from biochemical vendors unless otherwise noted.

[0060] The total extraction rate T of long-chain dibasic acid is calculated by the following formula, where V is the volume (L) of the clear fermentation liquid after removing unreacted alkanes by membrane filtration of the fermentation liquid of long-chain dibasic acid, M is the dry weight (g) of the extracted long-chain dibasic acid, and C is the concentration (g / L) of the long-chain dibasic acid in the lower tank.

[0061]

number

[0062] The molar conversion rate K of the mixed substrate is calculated by the following formula, where M represents the dry weight (g) of the extracted long-chain dibasic acid. 1 represents the mass of an alkane. 2 represents the mass (g) of lauric acid. m represents the molecular weight of the long-chain dibasic acid. m 1 represents the molecular weight of an alkane. 2 represents the molecular weight of lauric acid.

[0063]

number

[0064] The production intensity Q of long-chain dibasic acids is calculated by the following formula: where C is the concentration (g / L) of long-chain dibasic acids in the lower tank, and H is the fermentation time (h).

[0065]

number

[0066] In the examples of this application, the fermentation strain Candida tropicalis mutant PF-UV-56 was selected to ferment long-chain alkanes to produce long-chain dibasic acids. This mutant has been deposited in the China General Microbiological Culture Collection Center under the accession number CGMCC No. 0356.

[0067] [Preparation of fermentation seed culture] The strain maintained in slant culture was inoculated into four 5 L shake flasks to activate the strain (liquid holding volume of the shake flask was 300 mL). The strain was cultured at a culture temperature of 32°C and a shaking incubator speed of 200 rpm. After 48 hours of culture, the activated seed culture of 1.2 was used as the fermentation seed culture.

[0068] [Composition of fermentation medium] Sucrose: 20g / L Corn steep liquor: 0.8g / L ·Yeast extract: 2.0g / L Sodium chloride: 0.8g / L Potassium dihydrogen phosphate: 3.0g / L Magnesium sulfate: 1.2g / L ·Urea: 1.2g / L Ammonium sulfate: 1.5g / L Sodium acetate: 1.5g / L

[0069] Example 1 (1) 1650g of dodecane was prepared, and 330g of lauric acid was added. The resulting mixture was heated to 60°C and mixed and dissolved while stirring at 150 rpm. Next, 1980g of water was added. Next, while continuing to mix, solid sodium hydroxide was added to adjust the pH of the system to 8, and a mixed substrate was obtained. (2) The fermentation was carried out in a 15 L fermentation tank to produce long-chain dibasic acids. The total liquid retention volume was 12 L. The volume of the fermentation seed culture was 1.2 L. The fermentation temperature was 32°C. The aeration rate was 1.0 VVM. The stirring rate was 250 rpm. The pH was not adjusted from 0 to 24 hours after the start of fermentation. The pH was adjusted to 7.0 24 hours after the start of fermentation, and then increased by 0.15 every 24 hours. The alkane / lauric acid mixed substrate prepared in step (1) was added to the fermentation system in five batches. The mixed substrate was added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of fermentation. (3) The total fermentation time was 145 hours. After the completion of fermentation, the pH of the fermentation liquid was adjusted to 9.5, heated to 90°C, and maintained for 20 minutes. The demulsified fermentation liquid was subjected to solid-liquid separation by membrane filtration to obtain a clear aqueous fermentation liquid. The filtration conditions were a membrane pore size of 20 nm and a filtration temperature of 40°C. Concentrated sulfuric acid was added to the clear aqueous fermentation liquid to adjust the pH to 3. This caused the long-chain dibasic acid contained in the clear aqueous fermentation liquid to crystallize and be separated. The separated long-chain dibasic acid was filtered and dried using a plate and frame filter to obtain a crude product of the long-chain dibasic acid. The filtration conditions were a filtration pressure of 0.5 MPa and a filtration temperature of 30°C. The drying conditions were a temperature of 80°C and a drying time of 20 hours.

[0070] The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 152.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.4 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1914.5 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 62.4%.

[0071] Example 2 (1) 1900g of dodecane was prepared, and 190g of lauric acid was added. The resulting mixture was heated to 60°C and mixed and dissolved while stirring at 250 rpm. Next, 2090g of water was added. Next, while continuing to mix, solid sodium hydroxide was added to adjust the pH of the system to 7.5, and a mixed substrate was obtained. (2) The fermentation was carried out in a 15 L fermentation tank to produce long-chain dibasic acids. The total liquid retention volume was 12 L. The volume of the seed culture of the fermentation bacteria was 1.2 L. The fermentation temperature was 32°C. The aeration rate was 1.0 VVM. The stirring rate was 250 rpm. The pH was not adjusted from 0 to 24 hours after the start of the fermentation. The pH was adjusted to 7.0 24 hours after the start of the fermentation, and the pH was increased by 0.2 every 24 hours thereafter. The alkane / lauric acid mixed substrate prepared in step (1) was added to the fermentation system in five batches. The mixed substrate was added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of the fermentation. (3) The total fermentation time was 150 hours. After the completion of fermentation, the pH of the fermentation liquid was adjusted to 9, heated to 75°C, and maintained for 40 minutes. The demulsified fermentation liquid was subjected to solid-liquid separation by membrane filtration to obtain a clear aqueous fermentation liquid. The filtration conditions were a membrane pore size of 25 nm and a filtration temperature of 50°C. Concentrated sulfuric acid was added to the clear aqueous fermentation liquid to adjust the pH to 4. This caused the long-chain dibasic acid contained in the clear aqueous fermentation liquid to crystallize and be separated. The separated long-chain dibasic acid was filtered through a plate and frame and dried to obtain a crude product of the dibasic acid. The filtration conditions were a filtration pressure of 1.0 MPa and a filtration temperature of 20°C. The drying conditions were a temperature of 105°C and a drying time of 5 hours.

[0072] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 156.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.6 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1973.0 g; the extraction rate of the crude product of acid was 93%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 60.2%.

[0073] Example 3 (1) 1650g of dodecane was prepared, and 330g of lauric acid was added. The resulting mixture was heated to 60°C and mixed and dissolved while stirring at 150 rpm. Next, 5940g of water was added. Next, while continuing to mix, solid sodium hydroxide was added to adjust the pH of the system to 8, and a mixed substrate was obtained. (2) The fermentation was carried out in a 15 L fermentation tank to produce long-chain dibasic acids. The total liquid retention volume was 12 L. The volume of the fermentation seed culture was 1.2 L. The fermentation temperature was 32°C. The aeration rate was 1.0 VVM. The stirring rate was 250 rpm. The pH was not adjusted from 0 to 24 hours after the start of fermentation. The pH was adjusted to 7.0 24 hours after the start of fermentation, and the pH was increased by 0.15 every 24 hours thereafter. After 24 hours after the start of fermentation, the alkane / lauric acid mixed substrate prepared in step (1) was added to the fermentation system in a continuous feed mode. The flow rate was 66 g / hour. (3) The total fermentation time was 144 hours. After the fermentation was completed, the pH of the fermentation liquid was adjusted to 9.5, heated to 90°C, and maintained for 20 minutes. The demulsified fermentation liquid was subjected to solid-liquid separation by membrane filtration to obtain a clear aqueous fermentation liquid. The filtration conditions were a membrane pore size of 20 nm and a filtration temperature of 40°C. Concentrated sulfuric acid was added to the clear aqueous fermentation liquid to adjust the pH to 3. This caused the long-chain dibasic acid contained in the clear aqueous fermentation liquid to crystallize and be separated. The separated long-chain dibasic acid was filtered through a plate and frame and dried to obtain a crude product of the long-chain dibasic acid. The filtration conditions were a filtration pressure of 0.5 MPa and a filtration temperature of 30°C. The drying conditions were a temperature of 80°C and a drying time of 20 hours.

[0074] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 153.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.5 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1941.6 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 63.3%.

[0075] Comparative Example 1 The experiment was carried out as described in Example 1, except for the following changes: in step (1), the mixed substrate of alkane and lauric acid was not used, and only alkane was added in batches during the fermentation in step (2) (total mass: 1980 g). The total fermentation time was 158 hours. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 150.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1833 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 58.2%.

[0076] Comparative Example 2 The experiment was carried out as described in Example 1, except for the following changes: in step (1), the mixed substrate of alkane and lauric acid was not used, and in step (2), only lauric acid was added in batches (total mass: 1980 g). The fermentation time was 155 hours in total. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 15.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 12.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 169.2 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 6.3%.

[0077] Comparative Example 3 The experiment was carried out as described in Example 1, except for the following changes: in step (1), the mixed substrate of alkane and lauric acid was not used, and only alkane was added in batches during the fermentation in step (2) (total mass: 1848 g). The total fermentation time was 150 hours. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 141.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 12.9 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1709.8 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 58.2%.

[0078] Example 4 The experiment was carried out as described in Example 1, except for the following changes: in step (1), alkane and lauric acid were mixed under stirring at room temperature, 1980 g of water was added, and then solid sodium hydroxide was added under mixing to adjust the pH of the system to 8 to obtain a mixed substrate. The fermentation time was 144 hours in total. The fermentation results were as follows: After measurement and calculation, the fermentation concentration of long-chain dibasic acid in step (2) was 142.0 g / L. The clear fermentation liquid obtained after membrane filtration in step (3) was 13.2 L. The mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1762 g. The extraction rate of the crude product of acid was 94%. The molar conversion rate of the mixed substrate of alkane and lauric acid was 57.4%.

[0079] Example 5 The experiment was carried out as described in Example 1, except for the following changes: in step (1), alkane and lauric acid were dissolved under stirring at 60°C, and water was not added to make an aqueous solution, and the resulting mixture was directly used in the fermentation process in step (2) without adjusting the pH with alkali. The fermentation time was 144 hours in total. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 135.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1650 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 53.8%.

[0080] Example 6 The experiment was carried out as described in Example 1, except for the following changes: in step (1), alkane and lauric acid were dissolved under stirring at 60°C, and water was added to obtain an aqueous solution, which was directly used in the fermentation process in step (2) without adjusting the pH with alkali. The fermentation time was 145 hours in total. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 142.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.5 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1802 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 58.7%.

[0081] Example 7 (1) 1800g of dodecane was prepared, and 300g of lauric acid was added. The resulting mixture was heated to 60°C and mixed and dissolved while stirring at 150 rpm. Next, 4200g of water was added. Next, while continuing to mix, solid sodium hydroxide was added to adjust the pH of the system to 7.5, and a mixed substrate was obtained. (2) The fermentation was carried out in a 15 L fermentation tank to produce long-chain dibasic acids. The total liquid retention volume was 12 L. The volume of the fermentation seed culture was 1.2 L. The fermentation temperature was 32°C. The aeration rate was 1.0 VVM. The stirring rate was 250 rpm. The pH was not adjusted from 0 to 24 hours after the start of fermentation. The pH was adjusted to 7.0 24 hours after the start of fermentation, and then increased by 0.2 every 24 hours. The alkane / lauric acid mixed substrate prepared in step (1) was added to the fermentation system in five batches. The mixed substrate was added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of fermentation. (3) The total fermentation time was 153 hours. After completion of the fermentation, the long-chain dibasic acids were extracted in the same manner as in step (1) of Example 1.

[0082] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 160.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.6 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 2030.4 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 62.2%.

[0083] Example 8 (1) 1650g of dodecane was prepared, and 198g of lauric acid was added. The resulting mixture was heated to 60°C and mixed and dissolved while stirring at 150 rpm. Next, 1848g of water was added. Next, while continuing to mix, solid sodium hydroxide was added to adjust the pH of the system to 8, and a mixed substrate was obtained. (2) The mixture was fermented to long-chain dibasic acids in a 15 L fermentation tank. The total liquid retention volume was 12 L. The volume of the seed culture of the fermentation bacteria was 1.2 L. The fermentation temperature was 32°C. The aeration rate was 1.0 VVM. The stirring rate was 250 rpm. The pH was not adjusted from 0 to 24 hours after the start of the fermentation. The pH was adjusted to 7.0 24 hours after the start of the fermentation. Thereafter, the pH was increased by 0.2 every 24 hours. The alkane / lauric acid mixed substrate prepared in step (1) was added to the fermentation system in five batches. The mixed substrate was added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of the fermentation. (3) The total fermentation time was 142 hours. After the fermentation was completed, the long-chain dibasic acids were extracted in the same manner as in step (1) of Example 1.

[0084] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 148.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 12.9 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 2048.9 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 62.1%.

[0085] Example 9 The experiment was carried out as described in Example 1, except for the following changes: in step (1), a combination of tetradecane and myristic acid was used instead of the combination of alkane and lauric acid. The fermentation time was 144 hours in total. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 115.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1405.3 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 53.2%.

[0086] Example 10 The experiment was carried out as described in Example 1, except for the following changes: in step (1), a combination of hexadecane and palmitic acid was used instead of the combination of alkane and lauric acid. The fermentation time was 144 hours in total. The fermentation results were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 98.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1197.6 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 51.6%.

[0087] Example 11 (1) 1650g of dodecane was prepared and divided into 5 portions of 330g each. 33.0g, 36.3g, 39.6g, 42.9g, and 46.2g of lauric acid were added to each of the 5 portions to prepare mixtures of alkanes and lauric acid with different composition ratios (these mixtures are referred to as numbers 1 to 5). The mixtures of alkanes and lauric acid were pretreated in the same manner as in step (1) of Example 1. (2) Fermentation into long-chain dibasic acids was carried out in the same manner as in step (2) of Example 1. The alkane / lauric acid mixed substrates prepared in step (1) were added to the fermentation system in the order of numbers 1 to 5. The mixed substrates were added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of fermentation. (3) The total fermentation time was 140 hours. After the fermentation was completed, the crude product of long-chain dibasic acid was extracted in the same manner as in step (3) of Example 1.

[0088] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 149.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.0 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1820.8 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 63.0%.

[0089] Example 12 (1) 1650g of dodecane was prepared and divided into 5 portions of 330g each. 52.8g, 56.1g, 59.4g, 62.7g and 66.0g of lauric acid were added to each of the 5 portions to prepare mixtures of alkanes and lauric acid with different composition ratios (these mixtures are referred to as numbers 1 to 5). The mixtures of alkanes and lauric acid were pretreated in the same manner as in step (1) of Example 1. (2) Fermentation into long-chain dibasic acids was carried out in the same manner as in step (2) of Example 1. The alkane / lauric acid mixed substrates prepared in step (1) were added to the fermentation system in the order of numbers 1 to 5. The mixed substrates were added 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the start of fermentation. (3) The total fermentation time was 140 hours. After the fermentation was completed, the crude product of long-chain dibasic acid was extracted in the same manner as in step (3) of Example 1.

[0090] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 154.0 g / L; the clear fermentation liquid obtained after membrane filtration in step (3) was 13.4 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1939.8 g; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 64.2%.

[0091] Example 13 (1) 1650g of dodecane was prepared and divided into 5 portions of 330g each. 33.0g, 36.3g, 39.6g, 42.9g, and 46.2g of lauric acid were added to each of the 5 portions. The mixture was heated and dissolved in the same manner as in step (1) of Example 1. Water was then added to each portion until the total mass was 768g. Solid sodium hydroxide was further added to adjust the pH of the system to 7.5, and mixtures of alkanes and lauric acid with different composition ratios were obtained (the mixtures are referred to as numbers 1 to 5). (2) As described in step (2) of Example 1, the mixture was fermented to a long-chain dibasic acid. The alkane / lauric acid mixed substrate prepared in step (1) was continuously fed to the fermentation system at a constant flow rate in the order of numbers 1 to 5. Number 1 was fed 24 to 48 hours after the start of fermentation, number 2 was fed 48 to 72 hours after the start of fermentation, number 3 was fed 72 to 96 hours after the start of fermentation, number 4 was fed 96 to 120 hours after the start of fermentation, and number 5 was fed 120 to 144 hours after the start of fermentation. The flow rate was 32 g / hour. (3) The total fermentation time was 144 hours. After the fermentation was completed, the crude product of long-chain dibasic acid was extracted in the same manner as in step (3) of Example 1.

[0092] The results of fermentation were as follows: the fermentation concentration of long-chain dibasic acid in step (2) was 152.0 g / L; the clear fermentation broth obtained after membrane filtration in step (3) was 13.2 L; the mass of the crude product of long-chain dibasic acid obtained after acid precipitation was 1886.0; the extraction rate of the crude product of acid was 94%; the molar conversion rate of the mixed substrate of alkane and lauric acid was 65.3%.

Claims

1. A long chain composition comprising a long chain alkane and a long chain carboxylic acid, the weight ratio of the long chain alkane to the long chain carboxylic acid is from (2:1) to (20:1); The long chain alkane is one or more selected from the group consisting of n-dodecane, n-tetradecane, and n-hexadecane; The long-chain carboxylic acid is one or more selected from the group consisting of lauric acid, myristic acid, and palmitic acid; The long chain composition further comprises water; The pH of the long chain composition is 5 to 12; The mass of the water is 0.5 to 10 times the total mass of the long-chain alkane and the long-chain carboxylic acid. Long chain composition.

2. The long chain composition of claim 1, which satisfies one or more of the following (i) to (iii): (i) the mass of water is 1 to 3 times the combined mass of the long-chain alkane and the long-chain carboxylic acid; (ii) the weight ratio of the long chain alkane to the long chain carboxylic acid is from (5:1) to (10:1); (iii) The long chain composition is for use in fermentation.

3. 2. The long chain composition of claim 1, which satisfies the following (i) and / or (ii): (i) exists in the form of a liquid or a solid-liquid mixture at 32°C; (ii) The number of carbon atoms in the long-chain alkane is the same as the number of carbon atoms in the long-chain carboxylic acid.

4. 4. The long chain composition of claim 3, which satisfies the following (i) and / or (ii): (i) pH is 7.5 to 8.0; (ii) exists in the form of an aqueous liquid at 32°C.

5. A combined long chain composition, comprising n long chain compositions according to any one of claims 1 to 4, present independently of each other, The n is a positive integer from 2 to 40, The mass ratio of the long-chain alkane to the long-chain carboxylic acid in the i-th long-chain composition is R ｉ where i is any positive integer from 2 to n, and the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the first long-chain composition is R １ and the mass ratio of the long-chain alkane to the long-chain carboxylic acid in the n-th long-chain composition is R ｎ Then, R ｉ－１ / R ｉ ≧1, R １ / R ｎ > 1, Combined long chain compositions.

6. The combined long chain composition of claim 5, which satisfies one or more of the following (i) to (iv): (i) the n long chain compositions are packaged or compartmentalized independently of each other; (ii) n is a positive integer from 4 to 20; (iii) The above R ｉ－１ / R ｉ The value of is 1.0001 to 10; (iv) R above １ / R ｎ The value is 1.01 to 10.

7. The combined long chain composition of claim 5, which satisfies one or more of the following (i) to (vi): (i) the n long chain compositions are prepared separately; (ii) the weight ratios of any two of the n long chain compositions, calculated based on the amount of long chain composition, are the same or different; (iii) the weight ratio of any two of the n long chain compositions, calculated based on the amount of long chain alkane, is the same or substantially the same; (iv) n is a positive integer from 5 to 10; (v) R above ｉ－１ / R ｉ is between 1.5 and 2; (vi) R above １ / R ｎ The value is between 1.5 and 2.

8. 1. A method for producing a long chain composition comprising a long chain alkane and a long chain carboxylic acid, comprising: the weight ratio of the long chain alkane to the long chain carboxylic acid is from (2:1) to (20:1); The method includes at least a mixing step of mixing the long-chain alkane and the long-chain carboxylic acid in the mass ratio to obtain the long-chain composition; The long chain alkane is one or more selected from the group consisting of n-dodecane, n-tetradecane, and n-hexadecane; The long-chain carboxylic acid is one or more selected from the group consisting of lauric acid, myristic acid, and palmitic acid; The long chain composition further comprises water; The mass of the water is 0.5 to 10 times the total mass of the long-chain alkane and the long-chain carboxylic acid. Manufacturing method.

9. The method according to claim 8, which satisfies one or more of the following (i) to (vi): (i) the long chain alkane is n-dodecane; (ii) the long chain carboxylic acid is lauric acid; (iii) carrying out the mixing step with heating and stirring; (iv) The above mixing step is carried out while heating to 50 to 60°C; (v) The above mixing step is carried out while stirring at 150 to 250 rpm; (vi) the weight ratio of said long chain alkane to said long chain carboxylic acid is from (5:1) to (10:1).

10. In the mixing step, water and a pH adjuster are further added, the mass of the water is 0.5 to 10 times the total mass of the long-chain alkane and the long-chain carboxylic acid; The amount of the pH adjuster used is an amount that adjusts the pH of the long chain composition to 5 to 12. The method according to claim 8.

11. The method according to claim 10, which satisfies one or more of the following (i) to (v): (i) adding said water and said pH adjuster to said long chain composition under heating and stirring; (ii) adding the water and the pH adjuster to the long chain composition while heating and stirring, the heating temperature being 50-60°C; (iii) adding the water and the pH adjuster to the long chain composition while heating and stirring, the speed of the stirring being 150-250 rpm; (iv) the mass of the water is 1 to 5 times the combined mass of the long-chain alkane and the long-chain carboxylic acid; (v) The amount of the pH adjuster used is an amount sufficient to adjust the pH of the long chain composition to 7.5 to 9.

12. The method according to claim 10 or 11, which satisfies one or more of the following (i) to (iii): (i) the pH adjuster is one or more selected from sodium hydroxide and potassium hydroxide; (ii) the mass of the water is 1 to 3 times the combined mass of the long-chain alkane and the long-chain carboxylic acid; (iii) The amount of the pH adjuster used is an amount sufficient to adjust the pH of the long chain composition to 7.5 to 8.

0.

13. A method for producing a long-chain dibasic acid, comprising the following steps (1) and (2): (1) Producing a long chain composition by the method of claim 8 to provide the long chain composition of claim 1 or the combined long chain composition of claim 5 (collectively referred to as the long chain composition); (2) fermenting the long-chain composition in the presence of a fermentation bacterium and a fermentation medium to convert the long-chain composition into a long-chain dibasic acid; wherein the long-chain dibasic acid is one or more selected from the group consisting of dodecanedioic acid, tetradecanedioic acid, and hexadecanedioic acid; The fermentation bacteria is one or more kinds of yeast selected from the group consisting of the genera Candida, Cryptococcus, Neurospora, Hansenula, Pichia, Rhodotorula, Torulopsis and Trichosporon.

14. The method according to claim 13, which satisfies the following (i) and / or (ii): (i) the fermenting bacterium is of the genus Candida; (ii) The fermentation medium comprises: 20-28 g / L sucrose; 0.8-1.5 g / L corn steep liquor; 2.0-4.0 g / L yeast extract; 0.8-1.2 g / L sodium chloride; 3.0-3.5 g / L potassium dihydrogen phosphate; Magnesium sulfate at 1.2-1.8 g / L; 1.2-4.8 g / L urea; 1.5 to 2 g / L ammonium sulfate; and Sodium acetate at 1.5-1.8 g / L.

15. The fermentation bacterium is a mutant strain of Candida tropicalis, PF-UV-56 (deposited at the China General Microbiological Culture Collection Center, accession number: CGMCC No. 0356). The method of claim 14.

16. The method of claim 13, wherein the fermentation comprises the following steps (i) and (ii): (i) a preliminary step of mixing the fermentation bacteria and the fermentation medium and fermenting for 5 to 60 hours to obtain a fermentation raw material solution; (ii) a conversion step of adding the long-chain composition to the fermentation raw material solution and fermenting for 60 to 400 hours to convert the long-chain composition into the long-chain dibasic acid.

17. The method according to claim 16, which satisfies the following (i) and / or (ii): (i) In the preliminary step, fermentation is carried out for 20 to 30 hours; (ii) In the above conversion step, fermentation is carried out for 100 to 140 hours.

18. The method according to claim 13, which satisfies one or more of the following (i) to (vii): (i) adding a seed culture of the fermentation bacteria at 2 to 30% by volume of the total liquid retention volume; (ii) the reaction temperature of the fermentation reaction is 25 to 37°C; (iii) The fermentation reaction is carried out with stirring at 100 to 1000 rpm; (iv) the aeration rate of the fermentation reaction is 0.2 to 10.0 VVM; (v) the reaction time of the fermentation reaction is 65 hours or more; (vi) the dosage of the long chain composition is 100 to 1000 g per 1 L of total liquid holding volume; (vii) The fermentation medium is fed in an amount of 12-80 g / L of total liquid holding volume.

19. The method according to claim 18, which satisfies one or more of the following (i) to (vii): (i) adding a seed culture of the fermentation bacteria at 10-15% by volume of the total liquid retention volume; (ii) The reaction temperature of the fermentation reaction is 28 to 32°C; (iii) The fermentation reaction is carried out with stirring at 150 to 300 rpm; (iv) the aeration rate of the fermentation reaction is 0.5 to 1.0 VVM; (v) the reaction time of the fermentation reaction is 138 to 160 hours; (vi) the dosage of the long chain composition is 150-250 g per L of total liquid holding volume; (vii) The fermentation medium input is 37-42 g / L of total liquid holding volume.

20. The method according to claim 16, which satisfies the following (i) and / or (ii): (i) in the preliminary step, the long chain composition is not added and the pH of the fermentation reaction is allowed to self-regulate (i.e., the pH is not adjusted); (ii) At the start of the conversion step, the pH of the fermentation reaction is adjusted to 6.0-7.5, and thereafter the pH is increased by increments of 0.05-0.4 at intervals of 5-60 hours until the end of the fermentation reaction.

21. The method according to claim 20, which satisfies one or more of the following (i) to (iv): (i) within 24 hours of initiating the preliminary step, no long chain composition is added and the pH of the fermentation reaction is allowed to self-regulate; (ii) adjusting the pH of the fermentation reaction to 6.8-7.0 24 hours after the start of the preliminary step; (iii) increasing the pH by increments of 0.1 to 0.3; (iv) The pH is increased at intervals of 20 to 30 hours.

22. The method according to claim 16, which satisfies the following (i) or (ii): (i) adding said long chain composition in n batches in said converting step; The time intervals between two adjacent batches may be the same or different; The n is a positive integer from 2 to 40; (ii) In the conversion step, the long chain composition is added continuously depending on the reaction time.

23. The method of claim 22, which satisfies the following (i) and / or (ii): (i) the time interval between two adjacent batches is between 20 and 30 hours; (ii) The above n is a positive integer from 5 to 10.

24. 23. The process according to claim 22, which uses the combined long chain composition according to claim 5 and satisfies either (i) or (ii) below: (i) adding n of said long chain compositions in n batches in the order of a first long chain composition to an nth long chain composition; (ii) adding n of said long chain compositions sequentially according to a function R=f(t); During the ceremony, R is the mass ratio of the long chain alkane to the long chain carboxylic acid; t is the reaction time of the conversion step; f() represents any non-increasing function.

25. where f() represents any decreasing function, any monotonically decreasing function, or any linearly decreasing function. The method of claim 24.

26. Further comprising a step of separating the long chain dibasic acid from the reaction product of the fermentation reaction. The method of claim 13.

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