Leuconostoc mutant strain and constructing method and application method thereof

A technology of Leuconostoc and mutant strains, applied in the field of bacteria, can solve the problem of high cost and achieve the effect of overcoming the high cost

Inactive Publication Date: 2014-06-18
HEBEI UNIV OF TECH
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  • Abstract
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  • Claims
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a Leuconostoc mutant strain and its construction method and application method. The Leuconostoc mutant strain is a Leuconostoc mutant strain with dextran sucrase gene knockout. Recombination, by knocking out the gene to inactivate the dextran sucrase gen

Method used

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  • Leuconostoc mutant strain and constructing method and application method thereof
  • Leuconostoc mutant strain and constructing method and application method thereof
  • Leuconostoc mutant strain and constructing method and application method thereof

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Embodiment 1

[0034] The Leuconostoc MesenteroidesΔdtsl (Leuconostoc MesenteroidesΔdtsl) strain was constructed by inactivating the dextran sucrase gene of Leuconostoc by gene knockout. The steps are as follows:

[0035] The first step, the cloning of the partial sequence of Leuconostoc dextran sucrase gene

[0036]The Leuconostoc mesenteroides whose gene coding sequence length is more than 4kb, the preservation date is December 31, 2009, and it is preserved in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee (CGMCC), and the preservation number is CGMCC1.10327 ) dextran sucrase is divided into two segments and cloned and then spliced ​​into a part of the continuous sequence in the ORF. The specific operation steps are:

[0037] (1) Extraction of total DNA of Leuconostoc enterolis with deposit number CGMCC1.10327

[0038] Streak the strain with preservation number CGMCC1.10327 frozen at -80°C on the MRS solid plate and culture ove...

Embodiment 2

[0063]A Leuconostoc MesenteroidesΔdtsl (Leuconostoc MesenteroidesΔdtsl, deposit number: CCTCCM2013724) was used to ferment and produce mannitol.

[0064] The Leuconostoc Mesenteroides Δdtsl (Leuconostoc Mesenteroides Δdtsl, preservation number: CCTCCM2013724) strain was transferred to the MRS medium at 1% and cultured at 30°C on a shaker with a rotation speed of 120 rpm 20h, the metabolites were detected, and the comparison test proved that the mannitol yield of the Leuconostoc mesenteroides was 15.3% higher than that of the original Leuconostoc mesenteroides (preservation number: CGMCC1.10327), and the fructose part of the sucrose The conversion rate increased by 6.1%.

[0065] The application method of the above-mentioned Leuconostoc mutant strain, the composition of the MRS medium is: 2 grams of yeast extract powder, 20 grams of sucrose, 2 grams of ammonium citrate, 5 grams of sodium acetate, K 2 HPO 4 2 g, MnSO 4 ·H 2 O 0.039 g, Tween 80 0.4 ml and water 1000 ml, adjus...

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Abstract

The invention discloses a leuconostoc mutant strain and constructing method and application method thereof, relates to bacteria, is leuconostoc mutant strain knocked-out by dextransucrase gene, and is leuconostoc mesenteroides delta dtsl strain which is persevered by China Center for Type Culture Collection, the preservation date is 19th, Dec, 2013, and the preservation number is CCTCC NO.M2013724; the constructing method comprises the following steps: inactivating the dextransucrase gene of the leuconostoc through gene knock-out measurement and then constructing the leuconostoc mutant strain, the strain is transferred to an MRS culture medium in 1%, and then the stain is cultured on a shake bed with rotating speed of 120rpm for 20h at 30 DEG C, the mannitol yield of the leuconostoc mutant strain is improved by 15.3% in comparison with original leuconostoc mesenteroides, and the transfer rate of fructose in saccharose is improved by 6.1%.

Description

technical field [0001] The technical solution of the invention relates to bacteria, specifically a Leuconostoc mutant strain and its construction method and application method. Background technique [0002] Mannitol is widely used in pharmaceutical, food, chemical and electronic industries. Global mannitol consumption accounts for about 11% of the total polyols, about 150,000 tons per year. The United States is the largest consumer of mannitol. In other countries such as Europe, the amount used to produce chewing gum, buccal tablets, and chewable tablets is relatively large. my country's current consumption of mannitol is mainly used directly in the production of mannitol injections, pharmaceutical auxiliary additives and sugar-free food additives. Due to the rapid increase in the demand for mannitol in the food industry, especially chewing gum, the market capacity of mannitol has expanded rapidly. Mannitol is used in a large amount in the pharmaceutical industry. At pre...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12P7/18C12R1/01
Inventor 金红星成文玉张舟
Owner HEBEI UNIV OF TECH
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