Glucosamine synthase producing bacterium based on CRISPR-Cas9 technology and construction method and application of glucosamine synthase producing bacterium

A technology for producing bacteria and ammonia sugar, which is applied in the field of building and producing bacteria for ammonia sugar synthase, which can solve the problems of high GlcN accumulation concentration and difficulty in obtaining it, and achieve the effects of reducing inhibition, low production cost and short fermentation time
CN110387345AInactive Publication Date: 2019-10-29YANGZHOU RIXING BIO TECH +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
YANGZHOU RIXING BIO TECH
Publication Date
2019-10-29
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a glucosamine synthase producing bacterium based on a CRISPR-Cas9 technology and a construction method and application of the glucosamine synthase producing bacterium. The glucosamine synthase producing bacterium is obtained in the modes that a glucosamine acetylase gene gnal and a glucosamine synthase gene glms are connected to a carrier pET-28a, and then a recombinant plasmid is transferred into a double-gene deletion host bacterium obtained by knocking out a glucosamine metabolism gene naggE and a glucosamine metabolism gene mannX of an E.coli BL21(DE3) genome through the CRISPR-Cas9 gene editing technology. The recombinant bacterium is applied to fermentation culture, on a shake flask, 7.51 g / L of glucosamine can be accumulated, 3.73 g / L of acetylglucosamine canbe accumulated, and the total output is 11.24 g / L; and on a fermentor of 5 L, 7.30 g / L of the glucosamine can be accumulated, 10.53 g / L of the acetylglucosamine can be accumulated, and the total output is 17.83 g / L.
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Description

technical field

[0001] The invention relates to the technical field of gene knockout, in particular to a CRISPR-Cas9 technology-based ammonia sugar synthase-producing bacterium, a construction method and an application thereof. Background technique

[0002] Glucosamine (GlcN) has a good curative effect in the prevention and treatment of arthritis. Arthritis is mainly caused by joint damage and abnormal articular cartilage, which causes inconvenience to patients in daily activities and even leads to loss of bone and joint function. , Glucosamine stimulates chondrocytes through a certain mechanism in the body, effectively promotes the synthesis of collagen, and gradually repairs damaged articular cartilage. Glucosamine supplementation can promote the synthesis of glycoproteins to repair damaged cartilage, increase the lubrication between joints, and reduce the wear and tear between joints. In addition, glucosamine can also regulate the body's endocrine and adsorb on the intes...

Claims

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