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Method for inhibiting Lactobacillus plantarum by secreting and expressing plantaricin A from Saccharomyces cerevisiae

A technology of Lactobacillus plantarum and Lactobacillus plantarum, applied in the biological field, can solve the problems such as the heterologous expression of PlnA has not been found, and achieve the effect of environmental friendliness

Active Publication Date: 2018-05-01
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report on the heterologous expression of PlnA using Saccharomyces cerevisiae in the literature records that have been consulted so far

Method used

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  • Method for inhibiting Lactobacillus plantarum by secreting and expressing plantaricin A from Saccharomyces cerevisiae
  • Method for inhibiting Lactobacillus plantarum by secreting and expressing plantaricin A from Saccharomyces cerevisiae
  • Method for inhibiting Lactobacillus plantarum by secreting and expressing plantaricin A from Saccharomyces cerevisiae

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Experimental program
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Embodiment Construction

[0021] 1. Obtaining the target gene

[0022] 1. Take the preserved strain Lactobacillus plantarum ATCC BAA-793 and inoculate it into fresh MRS medium, and culture it overnight in an incubator at 30-37°C to recover the strain.

[0023] 2. The MRS medium formula used in step 1, 1L medium contains: peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, diammonium hydrogen citrate 2.0g, glucose (C 6 h 12 o 6 ·H 2 O) 20.0g, Tween-801.0mL, sodium acetate (CH 3 COONa·3H 2 O) 5.0g, dipotassium hydrogen phosphate (K 2 HPO 4 ·3H 2 O) 2.0g, magnesium sulfate (MgSO 4 ·7H 2 O) 0.58g, manganese sulfate (MnSO 4 ·H 2 O) 0.25g, the medium pH value is 6.2.

[0024] 3. Inoculate the revived Lactobacillus plantarum ATCC BAA-793 in fresh MRS medium, culture in a 30-37°C incubator for 12-16 hours, and collect the bacteria by centrifugation. The entire genome of Lactobacillus plantarum ATCC BAA-793 was obtained as a DNA template for PCR amplification using the Bacterial Whole Genome Ext...

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Abstract

The invention discloses a method for inhibiting lactobacillus plantarum by plantaricin A secreted and expressed by saccharomyces cerevisiae. The method utilizes gene engineering to modify saccharomyces cerevisiae so that the saccharomyces cerevisiae secretes plantaricin A (PlnA) and the plantaricin A can inhibit lactobacillus plantarum contamination in saccharomyces cerevisiae-based ethanol fermentation. The method belongs to the field of biology. Saccharomyces cerevisiae fermentation production of ethanol belongs to the field of biochemical engineering. Bioethanol industrial production is usually influenced by lactobacillus pollution and lactobacillus plantarum-caused damage is high. The method constructs saccharomyces cerevisiae engineering bacteria and makes the saccharomyces cerevisiae engineering bacteria secrete PlnA so that lactobacillus-caused pollution can be prevented and controlled. The engineering bacteria InvScI-plnA have good growth vigor, can successfully secrete PlnA, and have a concentration of about 20 micrograms per milliliter. After addition of the culture supernatant of the engineering bacteria into a lactobacillus culture system, a lactobacillus biomass change is measured and it is proved that the PlnA has obvious effects of inhibiting lactobacillus plantarum. Compared with the existing antibiotics, the PlnA is environmentally friendly, does not damage the human body and has a high latent business value and application potential.

Description

technical field [0001] The invention discloses a method for using genetic engineering to transform Saccharomyces cerevisiae so that it can secrete plantaricin A (PlnA), so as to inhibit plantaractobacillus infection in Saccharomyces cerevisiae ethanol fermentation, belonging to the field of biotechnology. Background technique [0002] In the industrial production of ethanol fermentation, bacterial contamination is a key problem that restricts the scale and sustainable development of fermentation for a long time. In the production of ethanol by fermentation of Saccharomyces cerevisiae, the main causes of bacterial contamination are equipment leakage and air pollution. After bacterial contamination occurs, the lactic acid content in the fermenter will increase, which shows that the main bacteria in the fermentation process are lactic acid bacteria. . Once infected, the conditions of the fermentation broth are suitable for the growth of Lactobacillus plantarum, and the growth ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/66C12N15/81C12P7/06
CPCY02E50/10
Inventor 李灏刘华擎王砚凤李思越
Owner BEIJING UNIV OF CHEM TECH