Bacillus subtilis and application thereof in biocatalytic production of L-lactic acid

A technology of Bacillus subtilis and biocatalysis, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, fixed on/in organic carriers, etc., can solve the problems of high energy consumption and high production costs, and reach the bottom line The effect of a wide range of objects and a strong ability

Inactive Publication Date: 2010-06-09
NANJING HUAZHOU PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The esterification method requires the treatment of high-concentration wastewater, which has high production costs and high energy consumption

Method used

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  • Bacillus subtilis and application thereof in biocatalytic production of L-lactic acid
  • Bacillus subtilis and application thereof in biocatalytic production of L-lactic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Preparation of free cells

[0051] Scrape 3 loops of Bacillus subtilis KR2 (CGMCC No.3242) slant strains with an inoculation loop in a sterile ultra-clean workbench, and inoculate them into the primary liquid seed medium (glucose: 15, yeast extract: 5, peptone: 5. Sodium chloride: 2, potassium dihydrogen phosphate: 1, dipotassium hydrogen phosphate: 1, magnesium sulfate: 0.5, caprolactam: 5, isovaleronitrile: 1, cobalt chloride: 10ppm, pH: 7.0, unit: g / L). At a temperature of 25°C and a rotating speed of 150rpm, the seed liquid of the first-grade Bacillus subtilis KR2 (CGMCC No.3242) can be obtained by shaking and culturing for 48 hours, and the first-grade seed liquid is transferred to the same The culture is continued in the sterilized medium, and the secondary seed liquid of Bacillus subtilis KR2 (CGMCC No. 3242) can be obtained after 48 hours.

[0052] Inoculate the secondary seed liquid of Bacillus subtilis KR2 (CGMCC No.3242) into 35L sterilized liqui...

Embodiment 2

[0053] Example 2 Preparation of Immobilized Cells

[0054] According to the mass ratio of 1.1% sodium alginate solution: 11% polyvinyl alcohol solution is 1:10, in a container, mix 1.1% sodium alginate solution and 11% polyvinyl alcohol solution and stir evenly , to prepare a carrier material solution. Then add the wet bacteria body of Bacillus subtilis KR2 (CGMCC NO.3242) into the carrier material solution to prepare a microbial mixed solution. The mass ratio of Bacillus subtilis KR2 (CGMCC NO.3242) wet thallus:carrier material is 1:5. Add the prepared microbial mixed solution dropwise to the calcium chloride solution with a mass concentration of 4% (adjust the pH value to 6.6 with sodium hydroxide solution) to carry out the first step of curing reaction of the microbial balls, and the curing reaction time is 0.5 hours. Use a net to pull out the microbial balls solidified in the first step, wash them three times with distilled water, and then put them into a saturated sodiu...

Embodiment 3

[0055] Example 3 Preparation of flocculated cell fluid

[0056] Take 3L of Bacillus subtilis KR2 (CGMCC NO.3242) fermentation broth and centrifuge at 3000rpm for 10min to obtain precipitated cells, wash once with 3L of deionized water and continue to centrifuge at 3000rpm for 10min, discard the supernatant to obtain precipitated wet cells. Bacillus subtilis KR2 (CGMCC NO.3242) precipitated wet cells were weighed and then washed with deionized water to prepare a bacterial suspension with a concentration of 35 g / L. Adjust the pH of the bacterial suspension to 7 at 30°C, add chitosan at a ratio of 2‰ (‰: g / 1000mL) to the bacterial suspension, and stir magnetically at 200 rpm for 10 minutes to prepare Bacillus subtilis KR2 (CGMCC NO.3242) flocculated cell fluid.

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Abstract

The invention belongs to the biochemical industry field and discloses a Bacillus subtilis and an application thereof in biocatalytic production of L-lactic acid. The method in biocatalytic production of L-lactic acid by using the Bacillus subtilis (the preservation number is CGMCC No.3242) comprises the steps as follows: taking immobilized cells or flocculated cells of the Bacillus subtilis (the preservation number is CGMCC No.3242) as a biocatalyst, catalyzing substrate lactonitrile to obtain L-lactic acid, dehydrating the reaction solution in a falling film evaporator, distilling and dehydrating by a molecular still to obtain the L-lactic acid with the product purity of over 95% and the product yield of over 90%. The invention has the advantages of simple technique operation, mild reaction condition, high utilization degree of the substrate, low energy consumption, high product purity, safety and environment protection, and wide application potential.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and relates to a bacillus subtilis and its application in biocatalytic production of L-lactic acid. Background technique [0002] Lactic acid (Lactic Acid, LA), also known as α-hydroxypropionic acid, molecular formula: CH 3 CH(OH)COOH is a natural organic acid and one of the three major organic acids. The lactic acid molecule contains an asymmetric carbon atom, so it has optical isomerism. The left-handed one is called L-lactic acid, the right-handed one is called D-lactic acid, and the mixture of the two is called D, L-lactic acid. Among them, only L-lactic acid can be absorbed and metabolized by the human body, and has good biological activity. With the strengthening of environmental awareness, the world's demand for biodegradable plastics is increasing. The polymer (polylactic acid) produced by processing L-lactic acid monomer is one of the most promising polymer materials. It is both It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/56C12N11/04C12R1/125
Inventor 杨寿海姜志忠孔国平芮忠南刘善和
Owner NANJING HUAZHOU PHARMA
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