A mannitol-producing Leuconostoc intestinalis mutant strain and its application method

A technology of Leuconostoc enterococcus and mutant strains, applied in the field of bacteria, can solve the problem of insufficient mannitol production rate

Active Publication Date: 2019-11-12
HEBEI UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The technical problem to be solved by the present invention is to provide a mannitol-producing Leuconostoc enteritidis mutant strain and its application method. Bacteria Δdtsl (Leuconostoc mesenteroides Δdtsl) was used as the starting bacterium, and molecular biology techniques were used to knock out the genes encoding D-lactate dehydrogenase and acetaldehyde dehydrogenase that consume NADH, and constructed glucan sucrase and D-lactate dehydrogenase Leuconostoc mesenteroides Δdts1ΔD-ldhΔaldh (Leuconostoc mesenteroidesΔdtslΔD-ldhΔaldh) strain with the knockout of enzyme and acetaldehyde dehydrogenase genes, that is, the preservation number is CCTCC No: M2016638, overcomes the existing The defect that the yield of mannitol produced by using sucrose as substrate in Leuconostoc fermentation technology is still not high enough

Method used

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  • A mannitol-producing Leuconostoc intestinalis mutant strain and its application method
  • A mannitol-producing Leuconostoc intestinalis mutant strain and its application method
  • A mannitol-producing Leuconostoc intestinalis mutant strain and its application method

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Embodiment 1

[0029] To construct a mutant strain of Leuconostoc enterococcus with dextran sucrase, D-lactate dehydrogenase and acetaldehyde dehydrogenase gene knockout, the specific steps are as follows:

[0030] In the first step, Leuconostoc enteritidis Δdtsl (CCTCC M2013724) was used as the starting bacterium to construct a mutant strain of Leuconostoc enteritidis with knockout of dextran sucrase and D-lactate dehydrogenase genes:

[0031] (1.1) Cloning of the partial sequence of the D-lactate dehydrogenase gene of Leuconostoc enterolis:

[0032] Using chromosomal DNA as a template, the Leuconostoc mesenteroidesΔdtsl (Leuconostoc mesenteroidesΔdtsl) strain with a coding sequence length of 996bp was cloned, and the preservation date was December 29, 2013. CCTCC) preservation, the preservation number is the partial continuous sequence of D-lactate dehydrogenase gene of CCTCC M2013724), and the specific operation steps are:

[0033] (1.1.1) The preservation number is the extraction of tot...

Embodiment 2

[0078] Leuconostoc mesenteroides ΔdtslΔD-ldhΔaldh (Leuconostoc mesenteroidesΔdtslΔD-ldhΔaldh) strain, which was preserved in the China Center for Type Culture Collection (CCTCC), the preservation date is November 14, 2016, and the preservation number is CCTCC M2016638), that is, the present invention The specific steps for the fermentation application of a mannitol-producing Leuconostoc enterica mutant strain are as follows:

[0079] Leuconostoc mesenteroides Δdts1ΔD-ldhΔaldh (Leuconostoc mesenteroidesΔdtslΔD-ldhΔaldh) in a 250ml Erlenmeyer flask, which will be preserved in the China Center for Type Culture Collection (CCTCC) with the preservation date of November 14, 2016, and the preservation number is CCTCC M2016638 The bacterial strain is transferred to MRS medium at 1% by weight, and cultivated at 30°C for 20 hours on a shaker with a rotation speed of 120 rpm. The concentration of mannitol can reach 9.35 g / L, and the conversion rate of fructose in sucrose 93.5%.

[0080]...

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Abstract

The present invention relates to a mannitol-producing Leuconostoc enterococcus mutant strain and its application method, and relates to bacteria. Mutant strain of Leuconostoc, Leuconostoc enterolis Δdtsl ∆D‑ldh∆aldh ( Leuconostoc mesenteroidesΔ dtsl∆D‑ldh∆aldh ) strain, which was preserved in the China Center for Type Culture Collection (CCTCC), the preservation date is November 14, 2016, and the preservation number is CCTCC M2016638. The strain was transferred to MRS medium at 1% by weight, and cultured at 30°C for 20 hours on a shaker with a rotation speed of 120 rpm. The concentration of mannitol could reach 9.35 g / L, and the fructose part of The conversion rate is 93.5%.

Description

technical field [0001] The technical scheme of the invention relates to bacteria, specifically a mannitol-producing Leuconostoc enterococcus mutant strain and its application method. Background technique [0002] Mannitol is a kind of hexyl alcohol, which is widely used in the fields of medicine, food and plastics. [0003] At present, there are two main processes for the industrial production of mannitol in the world. The first is the seaweed extraction method: extracting 1 ton of mannitol requires about 13 to 15 tons of dried kelp. While producing alginate, the kelp soaking liquid after extracting iodine is extracted and concentrated for many times, impurities are removed, and ion exchange is performed. , Concentrated by evaporation, cooled and crystallized; the production process produces a large amount of waste water, high energy consumption, serious pollution, and low yield. The second is catalytic hydrogenation method: using sucrose or glucose as raw material, it is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/18C12R1/01
CPCC12P7/18C12N1/205C12R2001/01
Inventor 金红星王立东成文玉
Owner HEBEI UNIV OF TECH
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