Actinoplanes spp. genetic manipulation system

Abstract

An Actinoplanes spp. genetic manipulation system introduces a phi derivative plasmid and a pIJ101 derived replicating plasmid into Actinoplanes spp. In a mycelium conjugal transfer mode in order to realize gene expression and knockout. The Actinoplanes spp. strain genetic manipulation system is established and systemically optimized, and a genetic manipulation tool suitable for like strains is developed and is successfully applied to accurate genome editing to improve the acarbose output and lays a foundation for realization of rational design and reconstruction of the Actinoplanes spp. strain. The system is adopted to reinforce expression of acarbose biosynthesis gene cluster (acb cluster) in order to increase of the final fermentation output of acarbose by 35%.

Description

technical field

[0001] The invention belongs to biotechnology, and in particular relates to a genetic operation system of the acarbose-producing bacterium Actinoplanes spp. Background technique

[0002] Acarbose is a pseudotetrasaccharide of C7 cyclic polyols, which can compete with α-glucosidase and reduce the effect of α-glucosidase on disaccharides and oligosaccharides in small intestinal wall cells. And the hydrolysis of polysaccharides to reduce the production, absorption and utilization of glucose and fructose, so as to achieve the purpose of reducing postprandial blood sugar in patients with type 2 diabetes. Because of its good pharmacokinetic properties and low toxicity, it has become an ideal drug for the treatment of type Ⅱ diabetes, and its sales share has surpassed traditional hypoglycemic drugs such as sulfonylureas and biguanides.

[0003] At present, the industry mainly uses Actinoplanes spp. to ferment and produce acarbose. The gene cluster responsible for i...

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