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Application of enterococcus faecium in preparation of L-lactic acid

A technology of Enterococcus faecium and lactic acid, which is applied in the application field of Enterococcus faecium in the preparation of L-lactic acid, can solve the problems of no Enterococcus strains for fermentation and production of L-lactic acid, low optical purity of the product, and many by-products. Achieve the effects of stable fermentation performance of strains, rich sources of raw materials, and less by-products

Active Publication Date: 2013-12-18
SHANDONG FOOD & FERMENT IND RES & DESIGN INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The United States, the Netherlands, Brazil and other countries have begun to use Lactobacillus delbrueckii for L-lactic acid fermentation, while domestic lactic acid production is dominated by Rhizopus, with many by-products and low optical purity of the product, which is far behind the international advanced level.
Domestic research on the production of L-lactic acid by bacterial fermentation mainly focuses on Lactobacillus, Streptococcus, and Bacillus, and there is no research on the production of L-lactic acid by Enterococcus strains There are no reports on the use of Enterococcus faecium strains to convert glucose into L-lactic acid by fermentation

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0045] Embodiment 1: Breeding of Enterococcus faecium producing high optical purity L-lactic acid

[0046] Inoculate the liquid seed medium with the starting strain Enterococcus faecium H-36, cultivate it to the middle logarithmic growth phase at 37°C, collect the bacteria by centrifugation, wash twice with sterile normal saline, and then add sterile normal saline to make a bacterial suspension. Make the cell concentration at 1×10 8 -10 9 pieces / ml. Take 10ml of the above-mentioned bacterial suspension and put it into a plate, put it under a 30W ultraviolet lamp to irradiate, the irradiation distance is 15cm, and the irradiation time is 3 minutes, 5 minutes, 7 minutes, 10 minutes. Bromocresol Purple-Calcium Carbonate Screening Plate for Calcium Lactate, incubate at 37°C for 1-2 days in the dark, pick a single colony that grows well and has a large discoloration transparent circle for preservation, and culture at 37°C for 1-2 days, after the slant is mature Inoculate the sha...

Embodiment 2

[0049] Example 2: Breeding of high yield and high optical purity L-lactic acid bacteria strain Enterococcus faecium CGMCC No.7274

[0050] The mutant strain H-U36 strain screened in Example 1 was inoculated into the liquid seed medium, cultivated to the middle logarithmic growth phase at 37°C, collected the bacteria by centrifugation, washed twice with sterile normal saline, and then added sterile normal saline to make Bacteria suspension, so that the cell concentration at 1 × 10 8 -10 9 pcs / ml, spare.

[0051] Preparation of nitrosoguanidine treatment solution: Weigh 20mg of nitrosoguanidine, place it in a 100ml sterile Erlenmeyer flask, add 2ml of acetone to dissolve it, then add 18ml of Tris buffer (pH6.0, 0.5mol / L) and mix well ,spare.

[0052] Take 10ml of the above-mentioned nitrosoguanidine treatment solution, add 10ml of bacterial suspension, incubate and shake at 30°C for 50-60 minutes, and take samples every 10 minutes. After sampling, first dilute 1000 times to ...

Embodiment 3

[0059] Example 3 Application of Enterococcus faecium in the preparation of L-lactic acid

[0060] The sequence of steps involved in applying the method is as follows:

[0061] (1) Strain selection: Enterococcus faeciums (Enterococcus faeciums) CGMCC N0.7274 was selected;

[0062] (2) Slant surface culture activation: inoculate the bacteria on the slant medium, and culture it statically for 30 hours at 37°C, and set it aside;

[0063] (3) Seed cultivation: Put the strain cultivated in step (2) under aseptic conditions into 80mL liquid seed medium (250mL Erlenmeyer flask) with an inoculation loop, set the rotation speed to 100 rpm, and the rotation radius On a 40mm shaker, culture at 37°C for 12 hours to obtain seed solution;

[0064] (4) Fermentation culture:

[0065] Shake flask fermentation: Inoculate the seed liquid in a shake flask (250mL Erlenmeyer flask) with 60 mL of fermentation medium at an inoculum volume of 3%, at 37°C, with a rotation speed of 100 rpm, and shake ...

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Abstract

The invention discloses application of enterococcus faecium in preparation of L-lactic acid, wherein the refereed bacterial strain is preserved in China General Microbiological Culture Collection Center on 5, March, 2013, and has the preservation number of CGMCCNo.7274. The application method comprises the following steps: (1) selecting bacterial strains; (2) cultivating and activating in solid media; (3) cultivating seeds; (4) carrying out fermentative cultivation; (5) detecting fermentation products, and the like. The method has the advantages of simplicity in operation, low cost, high yield of the target product L-lactic acid, high purity (over 98.6%), few common byproducts such as ethanol, fumaric acid, malic acid and the like, and wide application prospect.

Description

(1) Technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an application of Enterococcus faecium in preparing L-lactic acid. (2) Background technology [0002] Lactic acid (lactic acid), also known as a-hydroxypropionic acid and 2-hydroxypropionic acid, contains an asymmetric carbon atom in its molecule, so lactic acid molecules have optical isomers. L-lactic acid is its left-handed form, which naturally exists in animals, plants, microorganisms and human bodies. It is the only type of lactic acid that can be decomposed and utilized by humans and mammals. [0003] L-lactic acid has no toxic side effects on the human body, is easily absorbed by the human body, and can directly participate in the metabolism in the body. Therefore, L-lactic acid and its derivatives are used as high-safety spices, preservatives, preservatives, sour agents, bactericides, and stabilizers. Thickeners, curing agents, nutritional supplements, hu...

Claims

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Application Information

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IPC IPC(8): C12P7/56C12R1/46
Inventor 徐慧刘建军李文婧孙文涛
Owner SHANDONG FOOD & FERMENT IND RES & DESIGN INST
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