Engineering strain for producing high-molecular-weight hyaluronic acid, construction method thereof and application of engineering strain in producing high-molecular-weight hyaluronic acid

Abstract

The invention provides an engineering strain for producing high-molecular-weight hyaluronic acid, a construction method thereof and application of the engineering strain in producing the high-molecular-weight hyaluronic acid. The engineering strain is bacillus subtilis, and the Latin scientific name of the engineering strain is Bacillus subtilis; the engineering strain is named as BH-2, and the preservation number of the strain is CGMCC No.16841; the engineering strain selects bacillus subtilis WB600 as a basic strain, a hyaluronic synthase hasA gene, a UDP-glucose dehydrogenase has B gene anda UDP-glucose pyrophosphorylase hasC gene are introduced into the genome of the engineering strain, the characteristic of the low protease activity of the engineering strain is maintained, the growthand metabolism of the engineering strain are controlled through temperature, the high-molecular-weight hyaluronic acid with the molecular weight of 6.9*106 Dalton is obtained, and the engineering strain has industrial application value.

Description

technical field [0001] The invention relates to a method for constructing hyaluronic acid engineering bacteria and the obtained engineering bacteria, belonging to the fields of genetic engineering and microorganisms. Background technique [0002] Hyaluronic acid (hyaluronic acid or hyaluronan, HA) is a high-molecular glycosaminoglycan present in cartilage tissue, joint fluid, and dermis and epidermis of skin tissue, and its market demand is increasing day by day. Hyaluronic acid is involved in many important biological processes and can be widely used in fields such as ophthalmology, plastic surgery and cosmetics. At present, hyaluronic acid is generally produced in the industry using type C streptococci with weakened hemolytic toxicity. However, due to its genetic instability and complicated treatment methods, the production of HA by Streptococcus as a fermentation strain is limited. With the development of synthetic biology, it is possible to produce HA using strains wit...

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Problems solved by technology

[0004] At present, there is a lack of an engineering bacterium that can produce high molecular weight and medium molecular weight hyaluronic acid respectively under certain culture conditions (such as temperature, etc.), and will not contain a large amount of protease and hemolysin

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