阅读说明：本技术 一种二元酸胺盐的提取纯化方法 (method for extracting and purifying diamine salt ) 是由 董筱雯 杨晨 秦兵兵 刘修才 于 2018-05-29 设计创作，主要内容包括：本发明公开了一种二元酸胺盐溶液的纯化方法,包括以下步骤：(1)将包括杂质的二元酸胺盐溶液固液分离,得上层清液；(2)将步骤(1)中得到的上层清液,经树脂吸附,得二元酸胺盐溶液。本发明直接针对含有多种杂质的二元酸胺盐进行分离纯化,操作简单,可以很好的去除菌体、色素等杂质,使二元酸胺盐溶液能够达到高质量的聚合级别,由此聚合得到的聚合物,其纯度、色度等性能能够和现有聚合级单体聚合得到的聚合物相媲美,且二元酸胺的收率高。(The invention discloses a purification method of a diamine salt solution, which comprises the following steps: (1) carrying out solid-liquid separation on the diamine salt solution containing impurities to obtain supernatant; (2) and (2) adsorbing the supernatant obtained in the step (1) by resin to obtain a diamine dibasic acid salt solution. The method directly separates and purifies the diamine salt containing various impurities, has simple operation, can well remove impurities such as thalli, pigments and the like, and ensures that the diamine salt solution can reach high-quality polymerization level, and the polymer obtained by polymerization has the performances of purity, chromaticity and the like which are comparable with those of the polymer obtained by polymerization of the existing polymer-grade monomer, and has high yield of the diamine.)

1. A method for purifying a diamine salt solution, the method comprising the steps of:

(1) carrying out solid-liquid separation on the diamine salt solution containing impurities to obtain supernatant;

(2) And (2) adsorbing the supernatant obtained in the step (1) by resin to obtain a diamine dibasic acid salt solution.

2. the method of claim 1, wherein: in the step (1), the diamine salt comprises: a salt of a C4-C6 aliphatic or aromatic diamine and a C4-C18 aliphatic or aromatic dicarboxylic acid; the structural formula of the diamine is as follows: -OOC (CH2) nCOO- + H3N- (CH2) m-NH3+, wherein 4. ltoreq. n.ltoreq.18 and 4. ltoreq. m.ltoreq.6;

In the step (1), the diamine dibasic acid salt solution is obtained after fermentation liquor of dibasic acid or treatment liquid thereof participates in a pentanediamine fermentation process or participates in a process of lysine fermentation-enzyme conversion to obtain pentanediamine; wherein the diamine salt solution comprises impurities, and the impurities at least comprise thalli; alternatively, the impurities comprise at least a pigment.

3. the method of claim 1, wherein: in the step (1), the solid-liquid separation method comprises one or more of filtration, centrifugation or ceramic membrane separation;

And/or, in the step (1), before the solid-liquid separation, heating the diamine salt solution containing impurities;

And/or, in the step (1), the temperature of the solid-liquid separation is above room temperature, preferably above 60 ℃, more preferably above 70 ℃, and most preferably 70-90 ℃.

and/or, in the step (1), the rotation speed of the centrifugation is 4000-5500rpm, preferably 4500-5000 rpm;

And/or, in the step (1), the centrifugation time is 1-10 min.

4. the method of any one of claims 1-3, wherein:

In the step (2), the adsorption method comprises the following steps: contacting the supernatant obtained in the step (1) with macroporous adsorption resin for adsorption;

and/or, in the step (2), the pore diameter of the macroporous adsorption resin is 2-10nm, preferably 3-8 nm; the macroporous adsorption resin comprises: macroporous adsorption resin with the pore diameter of 3-6nm, macroporous adsorption resin with the pore diameter of 6-8nm and macroporous adsorption resin with the pore diameter of 3-4nm, preferably comprises the following components: one or more of XDA-1G macroporous adsorbent resin, XDA-5 macroporous adsorbent resin and DA201-C macroporous adsorbent resin.

5. The method of claim 4, wherein: in the step (2), the method of contacting includes any one of the following methods:

The first method is as follows: uniformly mixing the supernatant with macroporous adsorption resin, and adsorbing; and/or the presence of a gas in the gas,

the second method comprises the following steps: and (3) passing the supernatant through a macroporous adsorption resin column for adsorption.

6. the method of claim 5, wherein: in the first mode, the dosage of the macroporous adsorption resin is as follows: the volume ratio of the supernatant to the macroporous adsorption resin is (0.5-7): 1, preferably (1-5): 1, more preferably (1-3): 1;

and/or, in the first mode, the adsorption time is 5-72h, preferably 12-48 h;

And/or, in the first mode, the temperature of adsorption is 10-95 ℃, preferably 20-60 ℃, preferably 30-50 ℃;

and/or, in the second mode, the feeding flow rate of the supernatant liquid is 0.5-5BV/h, preferably 0.5-2 BV/h.

And/or in the second mode, the macroporous adsorption resin is in the form of a macroporous adsorption resin column, and the height-diameter ratio of the macroporous adsorption resin column is (10-20): 1.

7. A solution of diamine salts prepared by the process of any one of claims 1 to 6.

8. an amine salt of a dibasic acid, which is a solution having a concentration of 0.1% (m/V) and an absorption cell thickness of 5cm and having a UV index of 0.8X 10-3 or less, preferably (0.3-0.8). times.10-3, or preferably (0.1-0.3). times.10-3, as measured at 279 nm.

9. A method for preparing amine salts of dibasic acids, the method comprising the steps of:

(A) The process for the purification of a diamine salt solution of any one of claims 1 to 7 to obtain a diamine salt solution;

(B) Separating the amine salt of dibasic acid from the amine salt solution of dibasic acid in step (a).

10. the method of claim 9, wherein: in step (B), the separation method comprises crystallization; the crystallization rate is 1 ℃/3-15 min;

the cooling crystallization comprises the following steps: cooling the diamine salt solution to 20-40 ℃ at the rate of 1 ℃/3-10min, and preserving heat for 0.3-5 h; cooling at 1 deg.C/6-15 min for 5-20 deg.C, and maintaining for 0.2-5 h;

The water content in the diamine salt solution before crystallization is 25-40%, and the percentage is the mass percentage of water in the diamine salt solution.

Technical Field

The invention relates to a method for extracting and purifying diamine salt.

Background

Amine salts of dibasic acids (also known as polyamide salts or nylon salts) are precursors for the synthesis of polyamides. Polyamide is generally prepared by mixing amine and dibasic acid serving as monomer raw materials to form dibasic acid amine salt and then polymerizing the dibasic acid amine salt and the dibasic acid amine salt to obtain a polymer; the preparation process has the premise that: the monomeric amine and the dibasic acid are both necessarily polymerization-grade products, so that the obtained polyamide has industrial application value.

However, the preparation of polymer-grade dibasic acid and amine is complex in process and high in cost. Taking dibasic acid as an example, most of the existing dibasic acids are prepared by a biological fermentation method, and the polymer-grade dibasic acid is extracted from fermentation liquor by various complicated extraction and purification means. In general, obtaining conventional polymer grade diacids from fermentation broths requires the following complex steps: the fermentation liquor is processed by demulsification (such as alkali demulsification or heating demulsification), standing to remove substrate (such as alkane), crystallization (such as acidification crystallization) to obtain coarse product crystallized liquid of binary acid containing a large amount of thalli, filtering supernatant to obtain binary acid filter cake containing thalli, low-temperature drying the filter cake, washing to remove water-soluble impurities, adding organic solvent for extraction (simultaneously decoloring), filtering to remove decolorant, thalli, inorganic salt and the like to obtain organic solvent containing binary acid, and crystallizing, filtering and drying the binary acid to obtain binary acid product with high purity.

in addition, the fermentation liquor contains various impurities such as thalli, culture medium, unfermented substrate, a large amount of inorganic salt, protein, water and the like, and the fermentation liquor coexists in multiple phases, has complex composition and thick medium, has higher operation requirements of each step, consumes a large amount of energy, has high cost and low yield.

on the basis, Chinese patent CN00110713.5 is improved. CN00110713.5 indicates that: the main problems of the method for preparing the amine salt of the dibasic acid by using the long-chain dicarboxylic acid produced by the conventional fermentation method are that: (1) the pretreatment process of the fermentation liquor is complicated, and particularly, the drying of the crude acid filter cake containing bacteria and the final drying step of the dicarboxylic acid product are carried out at a lower temperature, so that the drying period is long, and the energy consumption is high; (2) the used extraction solvent has higher boiling point and is dried at low temperature, the solvent contained in the dicarboxylic acid product is difficult to completely remove, and the preparation of the diamine salt of the dibasic acid is adversely affected; (3) dicarboxylic acid extraction and diamine salt preparation use two different types of solvents, and the regeneration and recovery of the solvents require two different sets of processes and equipment, which further complicates the process. Therefore, the steps of drying the crude acid filter cake containing bacteria in the pretreatment process and crystallizing, filtering and drying the dicarboxylic acid product in the extraction process are omitted, the same solvent is used in the solvent extraction process and the preparation of the diamine salt, the process is simplified, the operation period and the energy consumption are shortened, and the product cost is reduced.

However, the technical scheme of CN00110713.5 is to treat the fermentation broth of long-chain dibasic acid, and then add diamine to form salt, but omits some steps in the treatment process of long-chain dibasic acid.

the method still has the problems of complex operation, high cost and low yield.

Disclosure of Invention

The invention provides a method for separating and purifying diacid amine salt containing various impurities, which aims to solve the problems that the preparation method of polymer-grade diacid and amine monomers is complex in operation, high in cost and low in yield, and the cost of the obtained industrially applied polymerization product is very high in the prior art. The method of the invention is simple to operate and can obtain the high-quality polyamide polymerization precursor.

Considering the inventive concept of the present invention, the conventional thinking of the prior art firstly obtains the high-quality polymerized monomers, namely, the dibasic acid and the amine, and then the dibasic acid and the amine are subjected to salt formation and polymerization to obtain various polymers. The object of the present invention is to remove various impurities from a complex system by using a mixed solution of a dibasic acid containing various impurities and a fermentation solution of an amine (equivalent to a mixed solution obtained by two fermentations or a treatment solution thereof) to obtain a polymer grade, and then polymerizing the salt solution of the polymer grade to obtain various polymers. None of the prior publications mention such a concept.

In practice, such a method is not only difficult to conceive, but also difficult to implement, requiring a number of technical difficulties to be overcome. The amine salt of dibasic acid fermentation broth or its treated solution contains a large amount of impurities such as: thalli, protein, a culture medium, an unfermented substrate, a large amount of inorganic salt, pigment, sugar, water and the like, and not only comprises various impurities in the dibasic acid fermentation process, but also various impurities in the pentamethylenediamine production process, and the whole system comprises multiphase substances such as gas, oil, water, solid and the like, and certain specific impurities need to be removed in the complex system, so that the high purity is ensured, and meanwhile, the high yield is also needed, and the difficulty is very high. In order to solve the problems, the inventor researches for many years and finds that various impurities can be synergistically removed through specific process steps and conditions, and further impurities such as thalli, pigments and the like can be well removed within a specific parameter range, so that a diamine salt solution can reach a polymerization level, and the performances of the polymer obtained by polymerization, such as purity, chromaticity and the like, can be comparable to those of the polymer obtained by polymerization of the existing polymerization level monomer, thereby obtaining the technical scheme of the invention.

one of the objects of the present invention is to provide a method for purifying a diamine salt solution, comprising the steps of:

(1) Carrying out solid-liquid separation on the diamine salt solution containing impurities to obtain supernatant;

(2) And (2) adsorbing the supernatant obtained in the step (1) by resin to obtain a diamine dibasic acid salt solution.

Further preferable embodiments of the above-described technical means will be described below.

in a preferred embodiment of the present invention, the amine salt of dibasic acid may include: salts of aliphatic or aromatic diamines of C4-C6 and aliphatic or aromatic dicarboxylic acids of C4-C18. Both the amine group of the diamine and the carboxyl group of the dicarboxylic acid are located at the terminal groups.

in a preferred technical scheme of the present invention, in the amine salt of dibasic acid, the structural formula of the dibasic acid is: HOOC (CH2) nCOOH, where n is 4. ltoreq. n.ltoreq.18, preferably 6. ltoreq. n.ltoreq.18, and may be 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18; the amine has the structural formula: H2N- (CH2) m-NH2, where 4. ltoreq. m.ltoreq.18, preferably 4. ltoreq. m.ltoreq.6, i.e. m can be 4, 5 or 6; the structural formula of the diamine is as follows: -OOC (CH2) nCOO- + H3N- (CH2) m-NH3+, wherein 4. ltoreq. n.ltoreq.18 and 4. ltoreq. m.ltoreq.6.

For example: the amine salt of a dibasic acid may be: glutaric acid ethylenediamine succinate, adipic acid glutaric acid diamine, sebacic acid glutaric acid diamine, dodecanedicarboxylic acid glutaric acid diamine, and the like. Amine salts of diacids may also include amine salts of diacids containing aromatic structures, such as: pentamethylene terephthalate, and the like. In order to obtain copolymers having different properties, diamine salts of polyamides of different types or diamine salts of polyamides and a monomer to be polymerized may be obtained as required. For example: the amine salt of dibasic acid of the present invention may further include nylon 66 salt, caprolactam, 6-aminocaproic acid, and the like. The amine salt of a dibasic acid of the invention may also be a mixture of different amine salts of dibasic acids.

According to a preferable technical scheme of the invention, in the step (1), the diamine dibasic acid salt solution is obtained after fermentation liquor of dibasic acid or treatment liquor thereof participates in a pentanediamine fermentation process or a process of lysine fermentation-enzyme conversion to obtain pentanediamine.

In a preferred technical scheme of the invention, the diamine salt solution is prepared by a fermentation method or prepared by a fermentation method and an enzyme conversion method.

according to a preferable technical scheme of the invention, the dibasic acid is prepared by a fermentation method.

in a preferred embodiment of the present invention, the amine is prepared by fermentation, or by fermentation and enzymatic conversion.

According to a preferable technical scheme of the invention, after the diacid amine salt solution is prepared by a fermentation method, before the method is carried out, the polymer-grade diacid and the polymer-grade amine are obtained by various methods and then dissolved to obtain the diacid amine salt, but the polymer-grade diacid amine salt is directly prepared from the fermentation liquor.

The concentration of the diamine salt solution of the present invention is not particularly limited as long as the diamine salt can be uniformly dissolved in the solution. According to a preferable technical scheme of the invention, in the step (1), the content of the diamine salt in the diamine salt solution is 10-30%, preferably 12-20%, and the percentage is the mass percentage of the diamine salt solution.

in a preferred technical scheme of the present invention, in the step (1), the solution of diamine salt is: a solution obtained by mixing a fermentation liquid and/or a treatment liquid of dibasic acid obtained by fermentation and an amine fermentation liquid and/or a treatment liquid of amine obtained by fermentation and conventional in the field; or the following steps: a solution obtained by mixing a fermentation solution and/or a treated solution thereof of a dibasic acid obtained by fermentation and an enzyme-converted solution and/or a treated solution thereof of an amine obtained by fermentation and enzyme conversion, which are conventional in the art. The impurities include: bacteria, proteins, culture medium, unfermented substrate, inorganic salts, pigment, reducing sugar and water. The impurities include at least bacteria. The impurities include at least a pigment.

In the present invention, the reducing sugar includes: saccharides containing a free aldehyde or ketone group include, inter alia, glucose.

in a preferred embodiment of the present invention, in step (1), the solid-liquid separation method includes one or more of filtration, centrifugation, or ceramic membrane separation. The filtration may be filter aid filtration or membrane filtration, etc.

In a preferred technical scheme of the invention, in the step (1), before the solid-liquid separation, the pH of the diamine dibasic acid salt solution is 6-10, preferably 7-9.

in a preferred technical scheme of the invention, in the step (1), before the solid-liquid separation, the diamine dibasic acid salt solution containing impurities is heated. The heating temperature is 70-100 deg.C, preferably 80-95 deg.C. The heating time is 30-120 min.

In a preferred embodiment of the present invention, in the step (1), the temperature of the solid-liquid separation is room temperature or higher, preferably 60 ℃ or higher, more preferably 70 ℃ or higher, and most preferably 70 to 90 ℃.

In a preferred technical scheme of the invention, in the step (1), the rotation speed of the centrifugation is 4000-.

In a preferred embodiment of the present invention, in step (1), the centrifugation time is 1-10 min.

In a preferred embodiment of the present invention, in step (2), the adsorption method comprises: and (2) contacting the supernatant obtained in the step (1) with macroporous adsorption resin for adsorption.

According to a preferable technical scheme of the invention, in the step (2), the pore diameter of the macroporous adsorption resin is 2-10nm, preferably 3-8 nm; the macroporous adsorption resin comprises: macroporous adsorption resin with the pore diameter of 3-6nm, macroporous adsorption resin with the pore diameter of 6-8nm and macroporous adsorption resin with the pore diameter of 3-4nm, preferably comprises the following components: one or more of XDA-1G macroporous adsorbent resin, XDA-5 macroporous adsorbent resin and DA201-C macroporous adsorbent resin.

In a preferred embodiment of the present invention, in the step (2), the method of contacting includes any one of the following methods:

The first method is as follows: uniformly mixing the supernatant with macroporous adsorption resin, and adsorbing; and/or the presence of a gas in the gas,

The second method comprises the following steps: and (3) passing the supernatant through a macroporous adsorption resin column for adsorption.

among the above-described embodiments, the second embodiment is preferable, and has a more prominent effect in removing reducing sugars than the first embodiment.

In a preferred technical solution of the present invention, in the first mode, the dosage of the macroporous adsorbent resin is as follows: the volume ratio of the supernatant to the macroporous adsorption resin is (0.5-7): 1, preferably (1-5): 1, more preferably (1-3): 1;

in a preferred technical scheme of the invention, in the first mode, the adsorption time is 5-72 hours, preferably 12-48 hours;

in a preferred technical scheme of the invention, in the first mode, the adsorption temperature is 10-95 ℃, preferably 20-60 ℃, preferably 30-50 ℃;

According to a preferable technical scheme of the invention, in the first mode, the supernatant and the macroporous adsorption resin are uniformly mixed, and after adsorption, the adsorption is more sufficient through one or more modes of oscillation, stirring or standing.

in a preferred embodiment of the present invention, in the second embodiment, the flow rate of the supernatant is 0.5-5BV/h, preferably 0.5-2 BV/h.

in a preferred technical solution of the present invention, in the second mode, the macroporous adsorbent resin is in the form of a macroporous adsorbent resin column, and the height-diameter ratio of the macroporous adsorbent resin column is (10-20): 1.

In a preferred embodiment of the present invention, in the second aspect, after the adsorption, washing is performed; the washing is preferably carried out by using distilled water, ammonia water or an acid solution; the flow rate during the washing is preferably 1-10BV/h, more preferably 5-8 BV/h; the acid solution is preferably hydrochloric acid solution; the temperature of the washing is preferably 20 to 95 ℃, more preferably 60 to 90 ℃.

in a preferred embodiment of the present invention, in the step (2), after the adsorption, the resin is separated from the diamine salt solution. The obtained diamine salt solution can be directly used for polymerization to obtain various polymers.

It is a second object of the present invention to provide diamine salt solutions prepared by the above-described process.

A third object of the present invention is an amine salt of a dibasic acid, which has a solution having a concentration of 0.1% (m/V) and an absorption cell thickness of 5cm and a UV index of 0.8X 10-3 or less, preferably (0.3-0.8). times.10-3, or preferably (0.1-0.3). times.10-3, as measured at 279 nm.

the diacid amine salt is not prepared by mixing polymer-grade diacid and polymer-grade amine. According to the common knowledge in the art, in general, the polymer-grade dibasic acid is a dibasic acid with a total acid content of more than 98%, or a dibasic acid with a total acid content of more than 98.5%, or a dibasic acid with a total acid content of more than 99%, or a dibasic acid with a total acid content of more than 99.5%; the polymer-grade pentamethylene diamine is pentamethylene diamine with the purity of more than 98 percent, or pentamethylene diamine with the purity of more than 99 percent, and pentamethylene diamine with the purity of more than 99.5 percent, and the percentage is mass percent.

the amine salt of a dibasic acid as described above, which has a concentration of 0.1% (m/V) and an absorption cell thickness of 5cm, has a UV index of 0.8X 10-3, 0.75X 10-3, 0.7X 10-3, 0.65X 10-3, 0.6X 10-3, 0.55X 10-3, 0.5X 10-3, 0.45X 10-3, 0.4X 10-3, 0.35X 10-3, 0.3X 10-3, 0.25X 10-3, 0.2X 10-3, 0.15X 10-3, 0.1X 10-3, measured at 279 nm.

The fourth object of the present invention is to provide a method for preparing diamine salt, which comprises the following steps:

(A) obtaining a diamine salt solution according to the purification method of the diamine salt solution of the diacid as described above;

(B) separating the amine salt of dibasic acid from the amine salt solution of dibasic acid in step (a).

In a preferred embodiment of the present invention, in the step (B), the separation method comprises crystallization. The crystallization is preferably a temperature-reducing crystallization. The cooling rate of the crystallization is preferably 1 ℃/3-15 min.

in a preferred embodiment of the present invention, the temperature-reducing crystallization comprises the following steps: cooling the solution of diamine salt to 20-40 ℃ at the rate of 1 ℃/3-10min, and keeping the temperature for 0.3-5 h; then the temperature is reduced by 5 to 20 ℃ at the speed of 1 ℃/6 to 15min, and the temperature is preserved for 0.2 to 5 hours.

according to a preferable technical scheme of the invention, the water content in the diamine salt solution before crystallization is 25-40%, and the percentage is the mass percentage of water in the diamine salt solution. The water content can be achieved by rotary evaporation.

the method directly separates and purifies the diamine salt containing various impurities, has simple operation, can well remove impurities such as thalli, pigments and the like, and enables the diamine salt solution to reach a high-quality polymerization level, and the polymer obtained by polymerization has the performances of purity, chromaticity and the like which are comparable with those of the polymer obtained by polymerization of the existing polymer-grade monomer.

Detailed Description

according to one embodiment of the present invention, the diamine forming the diamine salt of the dibasic acid at least comprises pentamethylenediamine, that is, the diamine forming the diamine salt of the dibasic acid is pentamethylenediamine or a mixed diamine of pentamethylenediamine and one or more of the following diamines: butanediamine, hexanediamine, heptanediamine, octanediamine, nonanediamine, decanediamine, p-phenylenediamine, and o-phenylenediamine.

in a preferred embodiment, one of the components of the diamine diacid salt is pentanediamine containing impurities, which may be typically a pentanediamine fermentation broth, an enzyme conversion broth, a salt solution, or the like.

In the present invention, the pentamethylene diamine is not limited in source, and can be produced by any conventional biological method. For example, a reaction in which a pentamethylenediamine decarboxylase acts on lysine to obtain an enzyme-converted solution, and further to obtain an enzyme-converted solution containing pentamethylenediamine (see JP 200400114A); through a genetic technology, the expression of lysine decarboxylase is up-regulated or the lysine decarboxylase is expressed in a recombinant mode in a strain capable of generating lysine, the produced lysine can be synchronously converted into pentanediamine in the fermentation process, and pentanediamine fermentation liquor can be obtained through direct fermentation (refer to the construction of a one-step method for producing 1, 5-pentanediamine corynebacterium glutamicum genetic engineering bacteria, such as Takara and the like, China journal of bioengineering, 2010, 30 (8): 93-99), and the like.

In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

the detection method of the performance parameters related to the embodiments of the present invention is as follows:

1. The method for detecting the purity of the diamine salt solution comprises the following steps:

Characterizing by a UV index; the absorbance A was measured at 279nm using a KONICA MINOLTA CM-3600A apparatus, and the UV index was 0.1% (m/V) of the concentration of the dibasic acid amine salt and 5CM of the thickness of the cell.

2. The method for detecting the color (chroma) of the diamine salt solution comprises the following steps:

and the visual detection method refers to GB/T605-2006 for detection.

3. the detection method of the content of reducing sugar comprises the following steps:

detection was performed according to DNS colorimetry using a KONICA MINOLTA CM-3600A instrument.

4. detection method of polyamide color:

according to GB-T2409-1980 standard, detection was carried out using a KONICA MINOLTA CM-3600A device.

Preparation of example 1

Preparation of sebacic acid pentanediamine salt fermentation liquor

1. preparation of ammonium sebacate fermentation liquor

1.1 strain activation:

Inoculating glycerol tube strain of Candida in seed bottle filled with YPD culture, and shake culturing at 29 deg.C and 220rpm for 1 day under natural pH; YPD medium comprises: 20g/L glucose, 10g/L yeast extract powder and 20g/L peptone; pH7.0;

1.2 seed tank culture, preparing seed liquid:

inoculating seeds in a seed bottle into a seed tank filled with a seed culture medium, wherein the initial pH value of a fermentation system after inoculation is 6.0, the aeration ratio is 0.5vvm at 29 ℃, the tank pressure is 0.1MPa, a certain stirring speed is kept, the dissolved oxygen in the seed culture process is controlled to be more than or equal to 10%, the seed is cultured for 18-20h, and the OD620 is 15 after the mature seeds are cultured by 30 times;

the seed culture medium comprises: 20g/L sucrose, 8g/L corn steep liquor, 5g/L yeast extract, 8g/L KH2PO4, 3g/L urea, prepared with water and sterilized at 121 deg.C for 20 min; sterilizing urea alone at 110 deg.C for 15min, cooling, and mixing with sterilized other components;

1.3, fermentation:

inoculating the seed solution into the fermenter containing a fermentation medium comprising: 40g/L glucose, 3g/L potassium nitrate, 5g/L monopotassium phosphate, 4g/L ammonium sulfate and 1g/L magnesium sulfate;

The temperature is controlled to be 29 ℃ in the fermentation process, the ventilation ratio is 0.3vvm, the tank pressure is 0.1MPa (gauge pressure), a certain stirring speed is kept to control dissolved oxygen to be more than or equal to 10 percent, the pH is controlled to be 6.0 at the beginning of fermentation, the pH is controlled to be 4.0-5.0 within 18h at the beginning of fermentation, and the pH is controlled to be 5.0-8.0 after the fermentation is finished for 18 h; adding n-decaalkane as substrate for the first time when fermenting for 18h, and then adding substrate again when the substrate content in the fermentation liquor is lower than 2%; in the fermentation process, 0-5 h of ammonia water is continuously supplemented at a speed of 0.5g/h/L (ammonia content of 25%), 5-18 h of ammonia water is continuously supplemented at a speed of 2g/h/L (ammonia content of 25%), 18-48 h of ammonia water is continuously supplemented at a speed of 1.5g/h/L (ammonia content of 25%), 48-120 h of ammonia water is continuously supplemented at a speed of 0.4g/h/L (ammonia content of 25%), and the addition amount of ammonia water is 100 g/L.

the total fermentation period is 165 h; obtaining ammonium sebacate solution with the concentration of 120 g/L.

2. Preparation of sebacic acid pentanediamine salt fermentation liquor

2.1 seeding tank culture: a 10L fermentation tank (working volume is 5.5L), the fermentation strain is CIB132-3 (a construction method, see PCT application numbers of PCT/CN2015/094121, WO2017/079872A1 and PCT application numbers of 2017, 5 months and 18 days, particularly see examples 1-17 in the specification of the patent application, such as example 16 and example 17), the inoculation ratio is 2%, the aeration ratio is 0.4vvm, the temperature is 37 ℃, the rotating speed is 700rpm, the tank pressure is 0.10MPa, the pH value of ammonia water is controlled to be 6.5, and the fermentation tank is accessed after the bacterial concentration OD562 reaches 1.00; seed culture medium: KH2PO40.4%, MgSO4 & 7H2O 0.25.0.25%, MnSO4 & H2O 13.5.5 ppm, ammonium sebacate (prepared in the above item 1, added in the form of ammonium sebacate solution, in percentages that are percentages of the effective components ammonium sebacate) 0.58%, glucose 15%, corn steep liquor 0.27%, threonine 0.035%, leucine 0.025%;

2.2 fermentation tank culture: a 10L fermentation tank (working volume is 6L), wherein the fermentation strain is CIB132-3, the inoculation ratio is 20%, the aeration ratio is 0.4vvm, the temperature is 37 ℃, the rotating speed is 800rpm, the tank pressure is 0.10MPa, the pH value of ammonia water is controlled to be 6.5, a supplemented medium is fed after 5h of fermentation, and the fermentation period is 35 h; fermentation medium: KH2PO40.04%, MgSO4 & 7H2O 0.25.0.25%, MnSO4 & H2O 0.017.017%, ammonium sebacate 0.3%, glucose 3.5%, corn steep liquor 0.50%, and threonine 0.022%; a supplemented medium: 50% of glucose and 25% of ammonium sebacate.

The concentration of glutaric diamine sebacate in the final fermentation broth was 235.33 g/kg.

preparation of example 2

Preparation of dodecadioic acid-pentanediamine salt fermentation liquor

1. preparation of ammonium dodecanedioate fermentation liquor

1.1 strain activation:

Inoculating a glycerol tube strain of the candida in a seed bottle filled with YPD culture, and carrying out shake culture at the temperature of 29 ℃ and the rpm of 220 and the amplitude of 26-50 mm for 1 day, wherein the pH is natural; YPD medium comprises: 20g/L glucose, 10g/L yeast extract powder and 20g/L peptone; pH7.0-7.5;

1.2 seed tank culture, preparing seed liquid:

inoculating seeds in a seed bottle into a seed tank filled with a seed culture medium, wherein the initial pH value of a fermentation system after inoculation is 6.0, the aeration ratio is 0.5vvm at 29 ℃, the tank pressure is 0.1MPa, a certain stirring speed is kept, the dissolved oxygen in the seed culture process is controlled to be more than or equal to 10%, the seed is cultured for 18h, and the OD620 is 15 after the mature seeds are cultured by diluting 30 times; the seed culture medium comprises: 20g/L sucrose, 8g/L corn steep liquor, 5g/L yeast extract, 8g/L KH2PO4, 3g/L urea, and 15mL/L substrate, prepared with water, and sterilized at 121 deg.C for 20 min; sterilizing urea alone at 110 deg.C for 15min, cooling, and mixing with sterilized other components;

1.3, fermentation:

Inoculating the seed solution into the fermenter containing a fermentation medium comprising: 30g/L glucose, 2g/L potassium nitrate, 3g/L monopotassium phosphate, 1g/L ammonium sulfate and 0.5g/L magnesium sulfate;

Controlling the temperature to be 29 ℃, the ventilation ratio to be 0.5vvm and the tank pressure to be 0.1MPa (gauge pressure) in the fermentation process, keeping a certain stirring speed to control the dissolved oxygen to be more than or equal to 10 percent, controlling the pH to be 6.0 at the beginning of fermentation, controlling the pH to be 4.0-5.0 within 18h at the beginning of fermentation, and controlling the pH to be 5.0-8.0 after 18h of fermentation is finished; adding substrate n-dodecyl alkane for the first time when fermenting for 18h, and then adding substrate again when the substrate content in the fermentation liquor is lower than 5%; continuously adding ammonia water (ammonia content is 25%) at a speed of 0.7g/h/L in the fermentation process, and adding 100g/L ammonia water after the fermentation is finished, wherein the total adding amount of the ammonia water is 210 g/L;

the total fermentation period is 155 h; to obtain the ammonium dodecanedioate solution with the concentration of 180 g/L.

2. preparation of dodecadioic acid-pentanediamine salt fermentation liquor

2.1 seeding tank culture: a 10L fermentation tank (7L), wherein the fermentation strain is corynebacterium glutamicum (Corynebacterium glutamicum), the inoculation ratio is 2%, the aeration ratio is 0.8vvm, the temperature is 39 ℃, the rotation speed is 400rpm, the tank pressure is 0.05MPa, the pH value of ammonia water is controlled to be 7.0, and the fermentation tank is accessed after the bacterial concentration OD562 reaches 1.00; seed culture medium: KH2PO40.5%, MgSO4 & 7 H2O0.35%, MnSO4 & H2O 16ppm, ammonium salt of dodecanedioic acid (prepared in the above 1 and added in the form of ammonium salt solution of dodecanedioic acid in percentages that are percentages of the effective components of ammonium salt of dodecanedioic acid) 0.4%, glucose 5%, corn steep liquor 0.30%, threonine 0.045%, and leucine 0.025%;

(2) Culturing in a fermentation tank: a 10L fermentation tank (working volume is 7L), wherein the inoculation ratio is 20%, the aeration ratio is 0.8vvm, the temperature is 39 ℃, the rotating speed is 500rpm, the tank pressure is 0.05MPa, the pH value of ammonia water is controlled to be 7.0, a prepared feed supplement culture medium is fed after fermentation is carried out for 6h, the fermentation period is 40h, and lysine fermentation liquor is obtained, the lysine content in the lysine fermentation liquor is 31.8%, and the pH value is 6.7; fermentation medium: KH2PO40.04%, MgSO4 & 7H2O 0.30.30%, MnSO4 & H2O0.010%, ammonium salt of dodecanedioic acid 0.4%, glucose 2.0%, corn steep liquor 0.50%, and threonine 0.030%; a supplemented medium: 20% of glucose and 5% of ammonium salt of dodecanedioic acid.

(3) Adding a certain amount of lysine decarboxylase prepared as above into lysine fermentation liquor, wherein the ratio of the added weight of lysine decarboxylase (calculated according to the cell dry basis of the lysine decarboxylase) to the weight of lysine in the lysine fermentation liquor (calculated according to lysine dodecate) is 1: 255, the addition amount of the coenzyme 5' -pyridoxal phosphate is 0.2mmol/L based on the weight of the reaction system; decarboxylation reaction is carried out for 12 hours at 100rpm and 35 ℃, lysine is converted into pentanediamine, the lysine conversion rate is more than 99%, and the reaction is finished to form a solution of dodecanedioic acid-pentanediamine salt;

In the final dodecanedioic acid-pentamethylene diamine salt solution (enzyme conversion solution), the concentration of dodecanedioic acid-pentamethylene diamine salt was 270.63 g/kg; the sulfate ion concentration was 0.43 g/kg.