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Method for producing D-lactic acid and brood-cell lactobacillus special for the same

A technology of Bacillus sporogenes and lactic acid, applied in the field of producing D-lactic acid, can solve the problems such as the decrease of the optical purity of D-lactic acid, the degeneration of strains, the contamination of miscellaneous bacteria, etc., and achieve the effects of high vigor, high transformation rate and avoiding pollution.

Active Publication Date: 2008-05-07
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

However, the production of D-lactic acid by semi-continuous fermentation has problems such as strain degradation, contamination of bacteria, and decrease in the optical purity of D-lactic acid.

Method used

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  • Method for producing D-lactic acid and brood-cell lactobacillus special for the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1. Isolation, mutagenesis screening and identification of Lactobacillus inulinus (Sporolactobacillus inulinus) CASD CGMCC №2185

[0046] 1. Isolation and mutagenesis screening of the strain Sporolactobacillus inulinus CASD CGMCC №2185

[0047] Isolation and screening of bacterial strains: Collect the soil of an orchard in Pinggu, Beijing, weigh 5 grams and dissolve it in 100ml of normal saline, mix it well, dilute it with sterile water 100,000 times, spread it on MRS solid medium, and cultivate it at 37°C; after the colonies grow out , Some single colonies dissolve the calcium carbonate in the medium around the colony due to acid production, forming a transparent circle, pick a single colony with an obvious transparent circle, inoculate it in 10ml of fermentation medium, and culture it statically at 37°C for 72 hours. The detection method described in the embodiment is to detect the content of D-lactic acid in the fermentation broth. After multiple screenings, ...

Embodiment 2

[0064] Example 2, Utilize Sporolactobacillus inulinus (Sporolactobacillus inulinus) CASD CGMCC №2185 to ferment and produce D-lactic acid in a conical flask, cycle fermentation twice, and ferment for 48 hours each time, ferment at 35°C, and 90% of the Replace the fermentation broth with the same volume of fresh fermentation medium

[0065] The composition of each culture medium used in the present embodiment is as follows:

[0066] Slant medium: glucose 20 g / L, yeast extract 5 g / L, peptone 10 g / L, beef extract 10 g / L, dipotassium hydrogen phosphate 2 g / L, diammonium citrate 2 g / L, acetic acid Sodium 5 g / L, Tween 80 1 mL / L, magnesium sulfate heptahydrate 0.58 g / L, manganese sulfate tetrahydrate 0.25 g / L, calcium carbonate 15 g / L, agar powder 15 g / L, solvent is water; The pH of the slant medium is 6.5. It is sterilized at 115° C. for 20 minutes.

[0067] Seed medium: glucose 40 g / L, yeast extract 5 g / L, peptone 10 g / L, beef extract 10 g / L, dipotassium hydrogen phosphate 2 g / L,...

Embodiment 3

[0075] Example 3, using Sporolactobacillus inulinus (Sporolactobacillus inulinus) CASD CGMCC №2185 to ferment and produce D-lactic acid in a conical flask, cycle fermentation twice, and ferment and cultivate for 65 hours each time, ferment at 40°C, and 70% of the Replace the fermentation broth with the same volume of fresh fermentation medium

[0076] The composition of each culture medium used in the present embodiment is as follows:

[0077] The slant medium and seed medium are the same as in Example 2.

[0078] Fermentation medium: industrial glucose (Zibo Hongyu Industrial Sugar Co., Ltd.) 160 grams / liter, soybean meal hydrolyzate (soybean meal hydrolyzate preparation method is the same as Example 1) 150 grams / liter, corn steep liquor (Shanghai Xiwang Starch Sugar Co., Ltd.) 40 g / L, calcium carbonate 80 g / L, and water as the balance; the pH of the fermentation medium is 6. Sterilize at 115° C. for 20 minutes.

[0079] The method for producing D-lactic acid by fermentatio...

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Abstract

The invention relates to a method for producing D-lactic acid and the special bacterial strain, in particular to a method for producing D-lactic acid and the special Sporolactobacillus. The Sporolactobacillus of the invention is Sporolactobacillus inulinus CASD and the preserved number is CGMCC NO2185. The method for producing D-lactic acid utilizes Sporolactobacillus inulinus CASD CGMCC NO2185 to produce D-lactic acid by fermenting hemicontinuously. The concentration of the D-lactic acid obtained by the invention can reach 162 g / L, the percent conversion can reach 94.7 to 96.0 percent and the optical purity can reach above 98.0 percent, further, the optical purity can still keep 98 percent after circular fermentation repeatedly and the pollution of impurity can be avoided. The invention has the advantages of enabling to be used for producing D-lactic acid, saving the cost, improving productivity efficiency, therefore being suitable for application in manufacturing production.

Description

technical field [0001] The present invention relates to a method for producing D-lactic acid and its special bacterial strain, in particular to a method for producing D-lactic acid and its special Lactobacillus sporogenes. Background technique [0002] Lactic acid is an organic acid with simple structure, which widely exists in nature as a metabolic intermediate and metabolite. Polylactic acid produced by lactic acid polymerization has good biocompatibility and biodegradability, and can be used to produce surgical sutures, biological implants, biodegradable fibers, packaging materials and agricultural films, etc. There is a chiral center in the lactic acid molecule, which has optical activity and is a chiral molecule. According to the different optical activity, lactic acid can be divided into L-lactic acid, D-lactic acid and racemic lactic acid. Among them, high-purity D-lactic acid is mainly used in biodegradable plastics, pesticides and herbicides, and pharmaceutical in...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/56C12R1/01
Inventor 许平马延和赵博秦加阳于波王丽敏马翠卿闫淑芳周帅
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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