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Method for producing L-lactic acid by fermentation and bacterial strain using same

A technology of lactic acid and bacterial strains, applied in the field of microorganisms, can solve problems affecting the unit output of lactic acid fermentation

Inactive Publication Date: 2013-04-17
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In addition, in the production of lactic acid fermentation, high concentrations of glucose and lactic acid have a strong inhibitory effect on the growth of lactic acid bacteria and lactic acid synthesis, which is a technical bottleneck affecting the unit output of lactic acid fermentation

Method used

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  • Method for producing L-lactic acid by fermentation and bacterial strain using same
  • Method for producing L-lactic acid by fermentation and bacterial strain using same
  • Method for producing L-lactic acid by fermentation and bacterial strain using same

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Experimental program
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Effect test

Embodiment 1

[0045] (1) Amplification of the coding sequence of the sigma factor;

[0046] According to the genome sequence of Lactobacillus rhamnosus HN001contig00032 published by genbank, the following pair of primers were designed using OLIGO 6.0 primer design software:

[0047] LRsigma-up: 5'-ATGGCTGATAAAAAAACAGCAAC-3'

[0048] LRsigma-down: 5'-TTATTCAAGAAAATCCTTAAGCTGC-3'

[0049] In the 50 μL reaction system, the concentration of the above two primers is 1 pmol / μL; dNTP each 20 pmol / μL; Mg 2+ 15pmol / μL; Lactobacillus rhamnosus genomic DNA template 10pmol; Pfu DNA polymerase 2.5U. The PCR amplification conditions were: 95°C for 5 min; 94°C for 1 min, 57°C for 1 min, and 72°C for 2 min for 30 cycles; 72°C for 10 min.

[0050] The PCR product is electrophoresed with 0.9% agarose gel (J. Sambrook, third edition of molecular cloning experiment guideline, 2002 edition), and the result shows to obtain a specific fragment of about 1251bp, coincides with the target fragment size of expecte...

Embodiment 2

[0106] Embodiment 2 Lactic acid fermentation of the high-temperature-resistant, high-sugar high-yielding lactic acid bacterial strain of the present invention

[0107] a. Activated strain: prepare MRS medium, the tissue components of MRS medium are: peptone 10g / L, beef extract 10g / L, yeast powder 5g / L, K2HPO42g / L, ammonium citrate 2g / L, sodium acetate 5g / L L, glucose 20g / L, Tween 801mL, MgSO4 7H2O 0.58g / L, MnSO4.4H2O 0.25g / L; use 6mol / L NaOH to adjust the pH value of the MRS medium to pH 6.8, and sterilize at 121°C for 30min ; Inoculate the preserved Lactobacillus rhamnosus (Lactobacillus rhamnosus) HRO3 CCTCC NO: M2010094 strain into 100 mL of MRS medium, and culture it overnight at 42°C and 150 rpm in a shaker to obtain an activated strain;

[0108] b. Seed preparation: inoculate the activated strain in the MRS medium according to the inoculation amount of 5-10% (v / v), place it in a shaker at 42°C and 150 rpm for 16 hours, and complete the seed preparation;

[0109] c. Ferm...

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Abstract

The invention provides an L-lactic acid fementative producing strain and a method for producing L-lactic acid through fermenting the producing strain. In the invention, the L-lactic acid fementative producing strain is Lactobacillus rhamnosus with NO. of M2010094. The L-lactic acid fementative producing strain has the characteristic of high temperature resistance and high sugar resistance, and can greatly improve the yield of the L-lactic acid.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacterial strain for fermenting and producing L-lactic acid and a method for fermenting and producing L-lactic acid. Background technique [0002] Lactic acid (α-hydroxy-propionic acid) widely exists in the metabolism of human body, animals and microorganisms, and also exists in various foods closely related to people's life. Lactic acid can be used in food, beverage, pharmaceutical plastics, feed, pesticides, daily chemicals, papermaking and electronics industries. In the food and beverage industry, lactic acid is used as a sour agent, fortifier, and preservative, and is an absolutely safe additive. Lactic acid derivatives, such as calcium lactate, zinc lactate, and ferrous lactate are fortifiers for food, beverages, and health products, and lactic acid ethyl lactate is the main aroma component of many famous wines. In the pharmaceutical industry, lactic a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/56C12N1/20C12R1/225
Inventor 鲁明波余龙江张力曾翔卢正东
Owner HUAZHONG UNIV OF SCI & TECH