Method for synchronously fermenting normal alkane to produce mixed long-chain dicarboxylic acid by using microorganism
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A long-chain dibasic acid and simultaneous fermentation technology, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of no public patents for mixed long-chain dibasic acids
Active Publication Date: 2011-07-06
青岛智库生物科技有限公司
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Problems solved by technology
[0003] In the patent literature, there is no relevant published patent on the production of mixed long-chain dibasic acids by biosynthesis
Method used
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Embodiment 1
[0029] (1). Take an inoculation loop of ly-m bacteria, spread it on the wort solid slope of a φ15×180 large test tube, and cultivate it at 30°C for two days.
[0030] (2). Take one of the above strains, insert it into a 250 mL Erlenmeyer flask containing 30 mL of alkane seed medium, and cultivate it at 30° C. on a rotary shaker at 220 rpm for 46 hours. Alkane Seed Medium, KH 2 PO 48g / L, yeast extract 3g / L, corn steep liquor 3.5g / L, sucrose 20g / L, urea 3g / L, heavy wax 30mL / L, prepared with tap water, pH 5.0, sterilized at 110°C for 30 minutes.
[0031] (3) In a 500mL Erlenmeyer flask containing 15mL of fermentation medium, add 3.5mL of the above seed solution, and ferment on a rotary shaker at 220 rpm for 4 days, and adjust the pH to 7.5-8.0 with NaOH every 24 hours. The fermentation medium mixture contains KH 2 PO 4 8g / L, NaAC 4g / L, KNO 3 3g / L, yeast extract 2g / L, corn steep liquor 2.5g / L, NaCl 1g / L, urea 2g / L, sucrose 20g / L, penicillin 160 units / mL, and 200 # Light wa...
Embodiment 2
[0034] According to the method of example 1, just ferment substrate with normal undecane (nC 11 ) (purity 99%), the result undecane dibasic acid (DC 11 ) output is 50.5g / L, DC 11 98.5% purity.
Embodiment 3
[0036] According to the method of example 1, just ferment substrate with n-tetradecane (nC 14 ) (purity 98%), without aggravating wax, resulting in tetradecanedioic acid (DC 14 ) yield is 71.5g / L, DC 14 98.1% purity.
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Abstract
The invention discloses a method for biosynthesizing and producing long-chain Alpha, Omega-dicarboxylic acid, in particular relates to a high yielding method of a C11-C14 mixed long-chain dicarboxylic acid (DCm). A used microorganism is a mutant new strain ly-m of a Candida tropicalis. The method is characterized in that culture media using each single normal alkane of C11-C18 and light wax oil as substrates are respectively connected to strains of the microorganism at first; on the first stage, a system is controlled to mainly grow cells, and also produce a certain quantity of dicarboxylic acid; on the second stage, the fermentation system is controlled to mainly produce acid and also grow cells; and on the third stage, the system is controlled to only produce the acid without growing cells. When the method is used for fermenting 200# light wax oil (a normal alkane mixture of C11-C14) to produce the DC11-DC14 mixed long-chain dicarboxylic acid (DCm), the Candida tropicalis ly-m and the 200# light wax oil are fermented in a fermentation tank with a volume of 210m3 for 160 hours, and the content of DCm in a fermentation clear liquid can reach 175g / L.
Description
Technical field: [0001] The invention relates to a method for synchronously fermenting mixed n-alkanes with microorganisms to produce mixed long-chain dibasic acids, especially for converting light wax oil with high yield including C 11 -C 14 Dibasic acid mixed dibasic acid (DC m )Methods. Background technique: [0002] Long-chain dibasic acids are important raw materials for synthetic perfumes, nylon engineering plastics, hot-melt adhesives, coatings, lubricating oils, resins and medicines. Long-chain mixed dibasic acids (DC m ) is an important raw material for synthesizing high-performance copolymerized nylon engineering plastics, high-grade nylon hot-melt adhesives for clothing, high-grade lubricants and coatings. Nylon 12 is a high-grade nylon engineering plastic with the longest carbon chain, the best quality and the highest price developed in the world, with an annual output of 100,000 tons. Nylon 12 was put into production by German Huls Company and Swiss Emser C...
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