CN118126848A - Candida tropicalis and method for producing long-chain dibasic acid by candida tropicalis - Google Patents

Candida tropicalis and method for producing long-chain dibasic acid by candida tropicalis Download PDF

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CN118126848A
CN118126848A CN202211535051.5A CN202211535051A CN118126848A CN 118126848 A CN118126848 A CN 118126848A CN 202211535051 A CN202211535051 A CN 202211535051A CN 118126848 A CN118126848 A CN 118126848A
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candida tropicalis
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alkane
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樊亚超
张霖
师文静
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Sinopec Dalian Petrochemical Research Institute Co ltd
China Petroleum and Chemical Corp
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China Petroleum and Chemical Corp
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    • C12R2001/74—Candida tropicalis

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Abstract

The invention relates to candida tropicalis and a method for producing long-chain dibasic acid by the candida tropicalis, wherein the candida tropicalis (Candida tropicalis) DLY-0728 is provided, the yield of the long-chain dibasic acid produced by fermentation is higher in the presence of an emulsifier with higher concentration, and the candida tropicalis is preserved in China general microbiological culture collection center (CGMCC) No.20459 in the year 2020 and the month 27. Compared with the original strain, the strain bred by the method has the advantages that the yield of the dibasic acid is obviously improved, and the stress resistance of the strain to the environment is stronger.

Description

Candida tropicalis and method for producing long-chain dibasic acid by candida tropicalis
Technical Field
The invention belongs to the technical field of biochemical engineering, and particularly relates to candida tropicalis and a method for producing long-chain dibasic acid by using the candida tropicalis.
Background
The long chain dibasic acid (DCA) refers to aliphatic dibasic acid with more than 10 carbon atoms in a carbon chain, and is an important raw material for manufacturing high-grade spice, high-performance nylon engineering plastics, high-grade nylon hot melt adhesive, high-temperature electrolyte, high-grade lubricating oil, high-grade paint and coating, cold-resistant plasticizer, resin, medicine, pesticide and the like.
Long chain dibasic acids have long been obtained from natural vegetable oils by hydrolysis, oxidation chain scission or cleavage, such as castor oil by thermal alkaline cracking to give sebacic acid, and erucic acid by ozone oxidation to give tridecanedioic acid. However, the production process of long-chain dibasic acid by using natural vegetable oil is complex, and the production process is influenced by natural disasters due to unstable raw material supply, thereby limiting the yield of the long-chain dibasic acid and influencing the development and application of long-chain dibasic acid products. At present, long-chain dibasic acid is produced abroad mainly by a chemical synthesis method, but the synthesis condition is harsh, high temperature and high pressure are needed, the steps are complicated, and fireproof, explosion-proof and anti-virus equipment and the like are also needed. And the chemical synthesis method has low yield, high cost and serious environmental pollution.
The biotechnology is applied to the petrochemical field, and opens up a new way for producing long-chain dibasic acid. The microbial fermentation method for producing long-chain dibasic acid gradually becomes the main production method of long-chain dibasic acid from the 70 th century of 20 th, with the advantages of mild reaction conditions, strong reaction specificity and the like.
The fermentation of long chain dibasic acid at present generally adopts batch fermentation, and alkane is supplemented in the fermentation process. And the long-chain dibasic acid fermentation system is a typical gas phase (oxygen) -water phase (fermentation liquid) -oil phase (alkane) -solid phase (thallus) four-phase system, and the uniformity of the four-phase system is maintained to help improve the fermentation effect. In the actual production process, in order to uniformly mix the system, a method of increasing the stirring speed is generally adopted, but increasing the stirring speed increases the shearing force, thereby affecting the growth of the fermentation cells. Or an emulsifier can be added into the fermentation medium, CN102115768A discloses a method for producing hexadecane diacid by synchronously fermenting n-hexadecane by microorganisms, and a surfactant (Tween 60) is added in an amount of 0.1-2 g/L to enhance the emulsification effect of an oil phase. However, in general, the emulsifier is not a component required for microbial metabolism, and thus this way of adding the emulsifier can have an adverse effect on the normal microbial fermentation.
CN103805643a discloses a method for producing long-chain diacid, which is to enlarge and culture diacid strains to obtain seed liquid, enlarge and ferment the seed liquid, add emulsified alkane in the fermentation process, remove mycoprotein after the reaction is finished, and crystallize to obtain long-chain diacid products. The emulsified alkane is obtained by mixing long-chain alkane with water and carrying out microwave treatment. Improves the compatibility between the water phase and the oil phase in the fermentation system, and improves the alkane conversion rate and the yield of the dibasic acid product in the fermentation system. However, the preparation of emulsified alkanes requires the use of microwave treatment and has limited emulsification effects.
CN111100884a discloses a method for preparing long-chain diacid by fermentation, which comprises (1) culturing long-chain diacid zymophyte to obtain seed liquid; (2) Adding fatty acid and water into alkane, and performing ultrasonic treatment to obtain gel-like alkane; (3) And adding the pretreated alkane and seed liquid into a fermentation culture medium, and fermenting in a mode of interval pH value regulation until the fermentation is finished. The invention increases the alkane pretreatment procedure, improves the alkane conversion rate, reduces the alkane loss, realizes the one-time addition of alkane, and simplifies the technological process. The invention needs to pretreat alkane, and needs to use fatty acid, phospholipid and other foreign substances, thus increasing fermentation cost.
Currently, microorganisms used to convert n-alkanes or fatty acids and their derivatives to the corresponding long chain dibasic acids are based on candida tropicalis (Candida tropicalis). The breeding of the fermentation strain plays a key role in reducing the process cost and improving the product quality, but often has limited functions. The improvement of the strain is generally realized by different mutagenesis and genetic engineering means, and the applicability, stability, stress resistance and the capability of producing long-chain dibasic acid of the strain can be further improved, so that the strain is more suitable for industrial application.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides candida tropicalis and a method for producing long-chain dibasic acid by using the candida tropicalis. Compared with the original strain, the strain bred by the method has the advantages that the yield of the dibasic acid is obviously improved, and the stress resistance of the strain to the environment is stronger.
The candida tropicalis (Candida tropicalis) DLY-0728 provided by the invention has higher yield of long-chain dibasic acid produced by fermentation in the presence of a higher concentration emulsifier, and is preserved in China general microbiological culture collection center (CGMCC) No. 20459 in the year 2020, month 07 and 27.
The main morphological characteristics of candida tropicalis (Candida tropicalis) DLY-0728 provided by the invention are as follows: the bacterial colony has white color, wrinkled surface and irregular edge, and the individual bacterial strain is oval under microscope.
The candida tropicalis (Candida tropicalis) DLY-0728 provided by the invention has stronger environmental stress resistance, and can better utilize emulsified alkane to ferment and produce long-chain dibasic acid in the presence of an emulsifier with higher concentration, so that the adverse effect of the emulsifier is avoided. The emulsifier is typically an anionic surfactant, preferably a sulfonate type anionic surfactant, more preferably sodium dodecylbenzenesulfonate.
The invention also provides application of the candida tropicalis (Candida tropicalis) DLY-0728 in fermentation production of long-chain dibasic acid.
In the application of the invention, the fermentation production of long-chain dibasic acid specifically comprises the following steps:
(1) Preparing zymophyte seed liquid: inoculating the preserved candida tropicalis (Candida tropicalis) DLY-0728 into a seed culture medium for culture to prepare fermentation bacteria seed liquid;
(2) Inoculating the fermentation bacteria seed liquid into a fermentation medium for fermentation, and obtaining long-chain dibasic acid fermentation liquid after fermentation.
In the above application, the formula of the seed culture medium in the step (1) is as follows: 10 to 50g/L of carbon source, 0.5 to 5g/L of nitrogen source, 0.5 to 10g/L of inorganic salt, 0.1 to 0.5g/L of vitamin B 1 and 2 to 10 percent of alkane. Wherein the carbon source is at least one of glucose, galactose, sucrose, maltose, trehalose, melezitose, etc., preferably sucrose; the nitrogen source is at least one of yeast extract, corn steep liquor, urea, ammonium sulfate, ammonia water, etc.; the inorganic salt is one or more of sodium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate, ferric sulfate, etc. Further, the preferred seed medium formulation is: 10 to 50g/L of sucrose, 0.5 to 5g/L of corn steep liquor, 0.5 to 5g/L of yeast extract, 0.5 to 5g/L of urea, 0.5 to 10g/L of sodium chloride, 0.5 to 10g/L of monopotassium phosphate, 0.5 to 10g/L of magnesium sulfate, 0.1 to 0.5g/L of vitamin B 1 and 2 to 10 percent of alkane.
In the above application, the culturing conditions in step (1) are: the temperature is 25-40 ℃, preferably 28-35 ℃, the pH value is 4.0-9.0, the rotating speed is controlled to be 200-400 rpm, and the culture is carried out until the OD 620 reaches over 6.0.
In the above application, the formula of the fermentation medium in the step (2) is as follows: 10 to 50g/L of carbon source, 0.5 to 10g/L of nitrogen source, 0.5 to 10g/L of inorganic salt, 0.1 to 0.5g/L of vitamin B 1 and 15 to 40 percent of alkane. Wherein the carbon source is at least one of glucose, galactose, sucrose, maltose, trehalose, melezitose, etc., preferably sucrose; the nitrogen source is at least one of yeast extract, corn steep liquor, urea, ammonium sulfate, ammonia water, etc.; the inorganic salt comprises one or more of sodium chloride, potassium chloride, sodium acetate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate, ferric sulfate, etc. Further, the preferred fermentation medium is formulated as follows: 10 to 50g/L of sucrose, 0.5 to 10g/L of corn steep liquor, 0.5 to 10g/L of yeast extract, 0.5 to 10g/L of urea, 0.5 to 10g/L of ammonium sulfate, 0.5 to 10g/L of sodium acetate, 0.5 to 10g/L of sodium chloride, 0.5 to 10g/L of monopotassium phosphate, 0.5 to 10g/L of magnesium sulfate, 0.1 to 0.5g/L of vitamin B 1 and 15 to 40 percent of alkane
In the application, the inoculation amount of the fermentation bacteria seed liquid in the step (2) is 5-20% of the volume of the fermentation medium.
In the above application, further, an amount of an emulsifier is added to the fermentation medium, wherein the emulsifier is generally an anionic surfactant, preferably a sulfonate type anionic surfactant, specifically at least one of alkylbenzenesulfonate, alkylsulfonate, fatty acid sulfoalkyl ester and the like, preferably sodium dodecylbenzenesulfonate, and the amount of the emulsifier is 10-500 mg/L, preferably 10-200 mg/L.
In the application, the fermentation culture conditions in the step (2) are as follows: the temperature is 25-40 ℃, preferably 28-35 ℃, the pH value is 4.0-9.0, the rotating speed is controlled to be 200-600 rpm, and the air flux is 0.3-2.0 vvm.
In the application, the step (2) regulates and controls the pH in the fermentation process, and the specific regulation and control modes are as follows: controlling the pH value of the system to be 4.0-7.0 within 0-24 h; and controlling the pH value to be 7.0-9.0 in 24-144 h, namely controlling the pH value of the system to be increased by 0.2 every 24h from 24h until 144h fermentation is finished.
In the above application, the alkane in step (1) and step (2) includes one or more of normal alkane of C10-C18, linear saturated fatty acid ester, linear saturated fatty acid salt and the like.
In the application, the long-chain dibasic acid of the fermentation product is one or more of C10-C18 straight-chain saturated dibasic acids.
Compared with the prior art, the invention has the following beneficial effects:
(1) The invention takes candida tropicalis CGMCC No. 0356 as an initial strain, and obtains candida tropicalis (Candida tropicalis) DLY-0728 by ultraviolet mutagenesis and chlorpromazine hydrochloride screening, the diacid output of the strain is obviously improved by more than 20g/L compared with the initial strain.
(2) The candida tropicalis DLY-0728 bred by the method has a certain alkane emulsifier tolerance capability, a certain amount of alkane emulsifier can be used in a fermentation system, the fermentation effect is improved, and the growth of thalli is not influenced.
Description of biological Material preservation
The candida tropicalis (Candida tropicalis) DLY-0728 provided by the invention is preserved in China general microbiological culture Collection center (CGMCC); preservation number: CGMCC No. 20459; preservation date: 2020, 07, 27; preservation address: the institute of microorganisms at national academy of sciences of China, national academy of sciences, no. 1, north Star West way, beijing, chao's area.
Detailed Description
The candida tropicalis of the present invention and its use are described in further detail below by way of examples. The embodiments and specific operation procedures are given on the premise of the technical scheme of the invention, but the protection scope of the invention is not limited to the following embodiments.
The experimental methods in the following examples, unless otherwise specified, are all conventional in the art. The experimental materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores.
In the embodiment of the invention, the concentration of the long-chain dibasic acid in the solution is determined by titration by adopting a sodium hydroxide standard solution, and the single-acid purity of the long-chain dibasic acid is analyzed by adopting gas chromatography.
The YPD plate medium formulation was: yeast extract 10g/L, peptone 20g/L, glucose 20g/L, agar powder 20g/L.
EXAMPLE 1 mutagenesis to obtain candida tropicalis DLY-0728
The strain PF-UV-56 of candida tropicalis (Candida tropicalis) in CN98121081.8 is taken as an original strain with the preservation number of CGMCC No. 0356, and the target strain candida tropicalis strain (Candida tropicalis) DLY-0728 is obtained by an ultraviolet mutagenesis and chlorpromazine hydrochloride screening method. The specific process is as follows:
(1) Culturing the mutant strain PF-UV-56 of the candida tropicalis in a seed culture medium to a logarithmic phase, centrifugally collecting thalli, washing twice with normal saline, scattering by glass beads, and filtering by a sterile filter membrane to obtain single-cell bacterial suspension. 5mL of single-cell bacterial suspension is taken in a culture dish, irradiated for 30 seconds at a distance of 15cm from a 30w ultraviolet lamp, then diluted by 10 -5 times, coated in YPD plate culture medium and cultured in the dark for 48 hours, and the mutagenized strain is obtained.
(2) The strain obtained by mutagenesis is coated on YPD plate culture medium containing chlorpromazine hydrochloride of 10mg/L, cultured for 48 hours, the strain growing on the culture medium is obtained and is used as a target strain, fermentation tests under different conditions are carried out on the target strain, and long-chain diacid strain with excellent fermentation performance is obtained by screening, namely candida tropicalis (Candida tropicalis) DLY-0728.
Example 2
The formula of the seed culture medium is as follows: sucrose 30g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 2% of n-dodecane.
The formula of the fermentation medium is as follows: sucrose 20g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; 1.0g/L of ammonium sulfate; 2.0g/L sodium acetate; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 25% of n-dodecane.
And (3) inoculating the preserved strain into an Erlenmeyer flask filled with a seed culture medium, culturing in a shaking table, controlling the temperature to be 30 ℃, controlling the rotating speed to be 200rpm, controlling the pH value in the culturing process, and obtaining the fermentation strain seed liquid after culturing for 36h and OD 620 to be 7.5.
The fermentation broth was inoculated into a 5L fermenter containing a fermentation medium at an inoculum size of 10%, a temperature of 30℃and a rotation speed of 500rpm, and an air flux of 1.0vvm. Controlling the pH value of the system to be 5.0 within 0-24 hours; starting from 24 hours, the pH value is controlled to be 7.5, and every 24 hours, the pH value of the control system is increased by 0.2 until 144 hours of fermentation is finished.
Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 187.8g/L, and the purity of monoacid is 99.6%.
Example 3
The formula of the seed culture medium is as follows: sucrose 30g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 2% of n-dodecane.
The formula of the fermentation medium is as follows: glucose 20g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; 1.0g/L of ammonium sulfate; 2.0g/L sodium acetate; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 25% of n-dodecane.
And (3) inoculating the preserved strain into an Erlenmeyer flask filled with a seed culture medium, culturing in a shaking table, controlling the temperature to 25 ℃, controlling the rotating speed to 250rpm, controlling the pH value in the culturing process, and obtaining the fermentation strain seed liquid after culturing for 36h and OD 620 to be 6.5.
The fermentation broth was inoculated into a 5L fermenter containing a fermentation medium at an inoculum size of 10%, a temperature of 25℃and a rotation speed of 600rpm and an air flux of 0.5vvm. Controlling the pH value of the system to be 5.0 within 0-24 hours; starting from 24 hours, the pH value is controlled to be 7.5, and every 24 hours, the pH value of the control system is increased by 0.2 until 144 hours of fermentation is finished.
After fermentation, sampling, detecting and analyzing, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 184.2g/L, and the purity of monoacid is 99.5%.
Example 4
The formula of the seed culture medium is as follows: sucrose 30g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 2% of n-dodecane.
The formula of the fermentation medium is as follows: maltose 20g/L; corn steep liquor 0.5g/L; yeast extract 0.5g/L; urea 1.0g/L; 1.0g/L of ammonium sulfate; 2.0g/L sodium acetate; sodium chloride 2.0g/L; 4.0g/L of monopotassium phosphate; 2.0g/L of magnesium sulfate; vitamin B 1 0.3.3 g/L; 25% of n-dodecane.
And (3) inoculating the preserved strain into an Erlenmeyer flask filled with a seed culture medium, culturing in a shaking table, controlling the temperature to 35 ℃, controlling the rotating speed to 300rpm, controlling the pH value in the culturing process, and obtaining the fermentation strain seed liquid after culturing for 36h and OD 620 to 7.0.
The fermentation broth was inoculated into a 5L fermenter containing a fermentation medium at an inoculum size of 10%, a temperature of 25℃and a rotation speed of 550rpm and an air flux of 2.0vvm. Controlling the pH value of the system to be 5.0 within 0-24 hours; starting from 24 hours, the pH value is controlled to be 7.0, and every 24 hours, the pH value of the control system is increased by 0.2 until 144 hours of fermentation is finished.
Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 181.3g/L, and the purity of monoacid is 99.4%.
Example 5
The difference from example 2 is that: sodium dodecyl benzene sulfonate is added into the fermentation medium with the dosage of 100mg/L. Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 195.5g/L, and the purity of monoacid is 99.7%.
Example 6
The difference from example 3 is that: sodium dodecyl benzene sulfonate is added into the fermentation medium with the dosage of 50mg/L. After fermentation, sampling, detecting and analyzing, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 192.4g/L, and the purity of monoacid is 99.6%.
Example 7
The difference from example 4 is that: sodium dodecyl benzene sulfonate is added into the fermentation medium with the dosage of 200mg/L. Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 190.6g/L, and the purity of monoacid is 99.8%.
Comparative example 1
The difference from example 2 is that: the fermentation strain adopts a candida tropicalis (Candida tropicalis) mutant strain PF-UV-56 in CN98121081.8, and the preservation number is CGMCC No. 0356. Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 151.3g/L, and the purity of monoacid is 99.5%.
Comparative example 2
The difference from example 5 is that: tween 80 was added to the fermentation medium at 100mg/L. Sampling, detecting and analyzing after fermentation is finished, wherein the concentration of long-chain dicarboxylic acid in the fermentation liquor is 185.3g/L, and the purity of monoacid is 99.5%.

Claims (18)

1. A candida tropicalis strain, which is characterized in that: the candida tropicalis (Candida tropicalis) DLY-0728 has higher yield of long-chain dibasic acid produced by fermentation in the presence of a higher concentration emulsifier, and is preserved in China general microbiological culture collection center (CGMCC) No. 20459 in the year 07 and the month 27 of 2020.
2. Candida tropicalis according to claim 1, characterized in that: the main morphological characteristics are as follows: the bacterial colony has white color, wrinkled surface and irregular edge, and the individual bacterial strain is oval under microscope.
3. Candida tropicalis according to claim 1, characterized in that: the candida tropicalis (Candida tropicalis) DLY-0728 has stronger environmental stress resistance, and can better utilize the emulsified alkane to ferment and produce long-chain dibasic acid in the presence of a higher concentration emulsifier.
4. An application of candida tropicalis (Candida tropicalis) DLY-0728 in fermentation production of long chain dibasic acid.
5. The use according to claim 4, characterized in that: the fermentation production of long chain dibasic acid comprises the following steps:
(1) Preparing zymophyte seed liquid: inoculating the preserved candida tropicalis (Candida tropicalis) DLY-0728 into a seed culture medium for culture to prepare fermentation bacteria seed liquid;
(2) Inoculating the fermentation bacteria seed liquid into a fermentation medium for fermentation, and obtaining long-chain dibasic acid fermentation liquid after fermentation.
6. The use according to claim 5, characterized in that: the formula of the seed culture medium in the step (1) is as follows: 10-50 g/L of carbon source, 0.5-5 g/L of nitrogen source, 0.5-10 g/L of inorganic salt, 0.1-0.5 g/L of vitamin B 1 and 2-10% of alkane; wherein the carbon source is at least one of glucose, galactose, sucrose, maltose, trehalose and melezitose, preferably sucrose; the nitrogen source is at least one of yeast extract, corn steep liquor, urea, ammonium sulfate and ammonia water; the inorganic salt is one or more of sodium chloride, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate and ferric sulfate.
7. Use according to claim 5 or 6, characterized in that: the formula of the seed culture medium is as follows: 10 to 50g/L of sucrose, 0.5 to 5g/L of corn steep liquor, 0.5 to 5g/L of yeast extract, 0.5 to 5g/L of urea, 0.5 to 10g/L of sodium chloride, 0.5 to 10g/L of monopotassium phosphate, 0.5 to 10g/L of magnesium sulfate, 0.1 to 0.5g/L of vitamin B 1 and 2 to 10 percent of alkane.
8. The use according to claim 5, characterized in that: the culture conditions of the step (1) are as follows: the temperature is 25-40 ℃, preferably 28-35 ℃, the pH value is 4.0-9.0, the rotating speed is controlled to be 200-400 rpm, and the culture is carried out until the OD 620 reaches over 6.0.
9. The use according to claim 5, characterized in that: the formula of the fermentation medium in the step (2) is as follows: 10-50 g/L of carbon source, 0.5-10 g/L of nitrogen source, 0.5-10 g/L of inorganic salt, 0.1-0.5 g/L of vitamin B 1 and 15-40% of alkane; wherein the carbon source is at least one of glucose, galactose, sucrose, maltose, trehalose and melezitose, preferably sucrose; the nitrogen source is at least one of yeast extract, corn steep liquor, urea, ammonium sulfate and ammonia water; the inorganic salt comprises one or more of sodium chloride, potassium chloride, sodium acetate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate and ferric sulfate.
10. Use according to claim 5 or 9, characterized in that: the formula of the fermentation medium is as follows: 10 to 50g/L of sucrose, 0.5 to 10g/L of corn steep liquor, 0.5 to 10g/L of yeast extract, 0.5 to 10g/L of urea, 0.5 to 10g/L of ammonium sulfate, 0.5 to 10g/L of sodium acetate, 0.5 to 10g/L of sodium chloride, 0.5 to 10g/L of monopotassium phosphate, 0.5 to 10g/L of magnesium sulfate, 0.1 to 0.5g/L of vitamin B 1 and 15 to 40 percent of alkane.
11. The use according to claim 5, characterized in that: the inoculation amount of the fermentation bacteria seed liquid in the step (2) is 5-20% of the volume of the fermentation medium.
12. The use according to claim 5, characterized in that: adding a certain amount of emulsifying agent into the fermentation medium, wherein the emulsifying agent is an anionic surfactant, preferably a sulfonate type anionic surfactant.
13. The use according to claim 12, characterized in that: the emulsifier is at least one of alkylbenzene sulfonate, alkyl sulfonate and fatty acid sulfoalkyl ester, preferably sodium dodecyl benzene sulfonate.
14. Use according to claim 12 or 13, characterized in that: the amount of emulsifier is 10-500 mg/L, preferably 10-200 mg/L.
15. The use according to claim 5, characterized in that: the fermentation culture conditions of the step (2) are as follows: the temperature is 25-40 ℃, preferably 28-35 ℃, the pH value is 4.0-9.0, the rotating speed is controlled to be 200-600 rpm, and the air flux is 0.3-2.0 vvm.
16. The use according to claim 5, characterized in that: the pH is regulated and controlled in the fermentation process in the specific regulation and control mode: controlling the pH value of the system to be 4.0-7.0 within 0-24 h; and controlling the pH value to be 7.0-9.0 in 24-144 h, namely controlling the pH value of the system to be increased by 0.2 every 24h from 24h until 144h fermentation is finished.
17. The use according to claim 5, characterized in that: the alkane of step (1) and step (2) comprises one or more of a C10-C18 normal alkane, a linear saturated fatty acid ester and a linear saturated fatty acid salt.
18. The use according to claim 5, characterized in that: the long-chain dibasic acid of the fermentation product is one or more of C10-C18 straight-chain saturated dibasic acids.
CN202211535051.5A 2022-12-02 2022-12-02 Candida tropicalis and method for producing long-chain dibasic acid by candida tropicalis Pending CN118126848A (en)

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