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A Strain of L-Ammonium Lactate Resistant Bacteria and Its Application

A technology of ammonium lactate and Lactobacillus casei, which is applied in the direction of bacteria, treatment of microorganisms with electricity/wave energy, biochemical equipment and methods, etc., can solve the problems of ammonium lactate production reduction, product inhibition, etc., and achieve great social significance and economic value , Relieve product inhibition, high conversion rate of sugar and acid

Inactive Publication Date: 2014-10-29
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the product inhibition caused by the accumulation of ammonium lactate in the fermentation process, so that the output of ammonium lactate is reduced, and a L-ammonium lactate tolerant strain is provided to greatly increase the output of L-ammonium lactate produced by fermentation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This example illustrates the method of low-energy nitrogen ion implantation mutagenesis and ammonium-resistant acclimatization of the starting strain of Lactobacillus casei.

[0045] The specific steps for the first step of low-energy nitrogen ion implantation mutagenesis screening and ammonium-resistant acclimatization are as follows:

[0046] (a) Bacterial suspension preparation: take the fresh slant of Lactobacillus casei LC-N235 cultured at 45-55°C for 48 hours, add 10 mL of sterile water, scrape and wash the colony and tilt it into a 250-mL test tube with small glass beads. Mix evenly on a mixer, break up the colonies, filter with three layers of degreasing gauze, check the filtrate under the microscope with simple staining, adjust the concentration of the bacteria solution until 0.1ml of the bacteria suspension is evenly spread on a 100cm culture dish without cell overlap.

[0047] (b) Low-energy nitrogen ion implantation mutagenesis: take 0.1mL of the bacterial s...

Embodiment 2

[0059] This example illustrates the genetic stability of L. casei LC-An226. The results of the subculture fermentation test are shown in Table 2

[0060] Table 2 Genetic stability of Lactobacillus casei LC-An226

[0061] Number of passages Production of L-ammonium lactate (g / L) 1 205 2 200 3 202 4 195 5 192 6 192

[0062] From the results of the genetic stability experiment, it can be seen that after 6 consecutive passages, the yield of L-ammonium lactate produced by the fermentation of the mutant strain Lactobacillus casei LC-An226 is relatively stable, and has good passage stability, which can be used as a production strain for further research and development.

Embodiment 3

[0064] This example illustrates the fermentation of the mutant Lactobacillus casei LC-An226 in a 5L fermenter to produce L-ammonium lactate.

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

[0066] Plate basic medium: glucose 0.5%, tryptone 1%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, light calcium carbonate 3%, agar 1.8%, the rest is distilled water, pH 6.5.

[0067] The slant medium is: glucose 0.5%, tryptone 1%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, light calcium carbonate 0.3%, agar 1.8%, the rest is distilled water, pH6.5.

[0068] The fermentation medium is: 22% glucose, 1% tryptone, 0.3% yeast extract, 0.15% potassium dihydrogen phosphate, and the rest is water, and neutralize it with 25% ammonia water to stabilize the pH of the medium at 6.5.

[0069] The screened mutant Lactobacillus casei LC-An226 was inoculated on the basic medium of the plate and cultured upside down at 50°C for 48 hours, and then...

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Abstract

The invention discloses an L-ammonium lactate tolerant bacterial strain and an application thereof in producing L-ammonium lactate through fermentation. According to the invention, low-energy nitrogen ions are injected in mutant original strains, and then the strains are screened by using a high-sugar tablet, a succinic acid tablet and an ammonium lactate tablet so as to obtain high-sugar-resistant and EMP approach reinforced strains which do not decompose and use a lactic acid. In a 3L fermentation tank, the strains are subjected to cultivation and acclimatization in a culture medium filled with critical-concentration ammonium lactate, and a bacteria liquid subjected to acclimatization of 72 hours is used as an original bacteria liquid of the next mutagenesis and acclimatization. The process is repeated until a target strain (Lactobacilluscasei) LC-An226 is screened. The strain has the characteristics of good ammonium lactate tolerance and high saccharic acid conversion rate. In a 5L fermentation tank, the output of L-ammonium lactate in the fermentation liquor reaches 206 g / L, and increased by 58.46% compared with the production of L-ammonium lactate by using original strains through fermentation, therefore, the strain has great social significance and economic values.

Description

technical field [0001] The invention relates to an L-ammonium lactate tolerant strain and its application in fermenting and producing ammonium lactate, belonging to the technical field of microbial fermentation. Background technique [0002] Ammonium lactate can be used in the pharmaceutical field to produce drugs for the treatment of skin diseases, and can also be used as an additive for animal feed. Ammonium lactate solution can be transformed into pure lactic acid solution through cation exchange resin. The direct esterification of ammonium lactate and ethanol can generate "green solvent" ethyl lactate. The most promising use of ethyl lactate is to take advantage of its non-toxic, good solubility, and non-volatile characteristics to replace many environmentally friendly solvents currently in use. Harmful solvents, such as: halogenated, ethers, and chlorofluorocarbon solvents that destroy the ozone layer. Ethyl lactate is mostly used in flavors, spices, food and brewing ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N13/00C12N15/01C12P7/56C12R1/245
Inventor 虞龙黄思思李玉燕刘媛张金丽陈晓双
Owner NANJING TECH UNIV
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