Bacillus coagulans and application thereof in mixed fermentation to produce L-lactic acid

A technology of Bacillus coagulans and mixed fermentation, which is applied in the direction of fermentation, bacteria, and microorganism-based methods, can solve the problems of low conversion rate and low optical purity of fermentation products, achieve high conversion rate of sugar and acid, shorten fermentation time, significant Effects of social significance and economic value

Inactive Publication Date: 2012-09-05
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rhizopus belongs to heterolactic acid fermentation with simple nutritional requirements, but the

Method used

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  • Bacillus coagulans and application thereof in mixed fermentation to produce L-lactic acid
  • Bacillus coagulans and application thereof in mixed fermentation to produce L-lactic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0041] This example illustrates the method of performing low-energy nitrogen ion implantation mutagenesis screening on the original strain of Bacillus coagulans.

[0042] The specific steps for the first step of low-energy nitrogen ion implantation mutagenesis screening are as follows:

[0043] (a) Preparation of single spore suspension: take the original starting strain of Bacillus coagulans BC-3 cultured at 40~45°C for 2~3 days and add 10 mL of sterile water to the fresh slant, scrape and wash the spores and pour them on the glass beads with certain amount Shake well in a 250mL conical flask for 20min (250rpm), break up the spore chains, filter through three layers of degreasing gauze, count the filtrate with a hemocytometer, and adjust the spore concentration by 10 6 individual / ml.

[0044] (b) Low-energy nitrogen ion implantation mutagenesis: take 0.1 mL of the spore suspension in step (a) and evenly spread it on a sterile plate, and perform low-energy nitrogen ion implan...

Embodiment 2

[0059] This example illustrates the genetic stability of high-yield strains Bacillus coagulans BC-N6262 and Lactobacillus casei LC-N235. The results of the subculture fermentation test are shown in Table 2:

[0060] Table 2

[0061]

[0062] From the experimental results, it can be seen that after 5 consecutive passages, the mutant strain Bacillus coagulans BC-N6262 has a relatively stable L-lactic acid production and good passage stability. In addition, the stability of Lactobacillus casei LC-N235 is also ideal. Both can be used as production strains for further research and development.

Embodiment 3

[0064] This example illustrates the production of L-lactic acid by mixed fermentation of Bacillus coagulans BC-N6242 and Lactobacillus casei LC-N235.

[0065] The medium formula described in this embodiment (% is mass percent):

[0066] Plate basic medium: glucose 5.0%, peptone 1.0%, beef extract 0.3%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 3.0%, agar 1.2%, the rest is water, pH 7.0.

[0067] Slant medium: glucose 0.5%, peptone 1.0%, beef extract 0.3%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 0.3%, agar 1.2%, the rest is water, pH 7.0.

[0068] Seed medium: glucose 6.0%, peptone 1.0%, yeast extract 1.0%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 3.0%, the rest is water, pH 7.0.

[0069] Fermentation medium: glucose 15%, peptone 0.05%, yeast extract 1.5%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 7.5%, the rest is water, pH 7.0.

[0070] Inoculate the sc...

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Abstract

The invention discloses bacillus coagulans BC-N6242 with collection number CCTCC (China Center for Type Culture Collection) M2012156. According to the invention, bacillus coagulans is mutated by low-energy nitrogen ion injection; and the screening of a high-glucose plate, a succinic acid plate and a pure-lactic acid plate and the lactobacillus casei'mixed fermentation' screening are adopted until the target strain is screened finally. The strain and the lactobacillus casei LC-N235 are proportionally mixed for fermentation. The seed culture solution is added to a 5L fermentation tank at a proportion of 2:1, and then fermentation and culture are performed for 48 hours; the output of the product L-lactic acid reaches 235g/L, which is improved by 45.1%, 57.7% and 14.6% as compared with that of separate fermentation of lactobacillus casei LC-N235, bacillus coagulans BC-3 and bacillus coagulans BC-N6242 respectively; the separate fermentation time of the bacillus coagulans is effectively shortened by 24 hours; and the bacillus coagulans and the application thereof in mixed fermentation to produce L-lactic acid provided by the invention have tremendous social significance and economic values.

Description

technical field [0001] The invention relates to a bacillus coagulans and its application in producing L-lactic acid by mixed fermentation, which belongs to the technical field of microbial fermentation. Background technique [0002] Lactic acid is an ancient and important organic acid. Lactic acid, lactate and its derivatives are widely used in medicine, medicine, feed, chemical industry and other fields. It is widely used as sour agent, preservative and reducing agent in food industry. Lactic acid, especially L-lactic acid has a strong bactericidal effect and can be directly used as a disinfectant in operating rooms, wards, laboratories and other places. L-lactic acid, L-sodium lactate, glucose, amino acid, etc. are compounded into infusion solution, which can treat acidosis and hyperkalemia. Lactic acid can be added to tobacco to maintain the humidity of tobacco and improve the quality of cigarettes. It can also be used to treat textile fibers, which can make it easy to...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P39/00C12P7/56C12R1/07C12R1/245
Inventor 李玉燕虞龙龚文静钱梦宇黄思思陈晓双
Owner NANJING UNIV OF TECH
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