Preparation method for high-molecular-weight hyaluronic acid (HA), and engineering bacterium

A hyaluronic acid, high molecular weight technology, applied in the field of bioengineering, can solve the problems of limited raw material sources, high production costs, cross-infection between virus species, etc.

Active Publication Date: 2014-08-20
JIANGSU HEVI BIOTECH CO LTD
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  • Abstract
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Problems solved by technology

This method mainly has the following disadvantages: the source of raw materials is limited, the extraction rate of the product is extremely low, the process procedure is complicated, the production cost remains high, and cross-infection between virus species may occur.

Method used

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  • Preparation method for high-molecular-weight hyaluronic acid (HA), and engineering bacterium
  • Preparation method for high-molecular-weight hyaluronic acid (HA), and engineering bacterium
  • Preparation method for high-molecular-weight hyaluronic acid (HA), and engineering bacterium

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Embodiment Construction

[0027] Below in conjunction with embodiment, following embodiment is illustrative, not limiting, can not limit protection scope of the present invention with following embodiment.

[0028] A vector containing a hyaluronan lyase gene, the vector contains a 1683bp gene fragment within the hylb gene (this fragment starts at 202bp downstream of the start codon ATG of the hylb gene and ends at 1310bp upstream of the stop codon TAG of the hylb gene) Insert the initial vector pSET4s (NCBI gene Bank: AB055650.1), construct the intermediate vector pSET4s::hylb, and then insert a gene fragment containing the chloramphenicol resistance gene cmr with a length of 1056bp into the DraIII position of the hylb gene fragment point, forming the vector pSET4s::hylb::cmr.

[0029] The content of the present invention includes: taking the temperature-sensitive plasmid pSET4s as the starting vector, inserting the 1683bp gene fragment inside the target gene hylb gene, and constructing the knockout pl...

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Abstract

The invention relates to a preparation method for high-molecular-weight HA and an engineering bacterium. According to the method, a coding hyaluronic acid lyase gene (hylb) in the genome of Streptococcus zooepidemicus ATCC39920 is knocked out, and a hyaluronic acid lyase gene (hylb)-knocked out vector pSET4s:: hylb:: cmr is constructed. The method provided by the invention subjects a recombinant bacterium in which the hylb gene is knocked out to fermentation and analysis, the molecular weight of HA is measured by using an intrinsic viscosity process, and HA with a molecular weight of 3.9 * 10<6> Da is obtained.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to knockout of the hyaluronic acid lyase gene in the genome of Streptococcus zooepidemicus, in particular to a method for preparing high-molecular-weight hyaluronic acid and engineering bacteria. Background technique [0002] High-molecular-weight hyaluronic acid plays an important role in the fields of cosmetics and medicine, and its preparation method is also a difficult and hot spot in current biotechnology. The method used to prepare high-molecular-weight hyaluronic acid is mainly animal extraction. [0003] Animal tissue extraction method, hyaluronic acid (HA) exists in almost all animal tissues, and the raw materials that can be used for production are mainly human umbilical cord, animal eyeball and chicken comb. The main process includes extraction, impurity removal, enzymatic hydrolysis, precipitation and separation. This method mainly has the following disadvantages: ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N1/21C12P19/26C12R1/46
Inventor 刘浩王震
Owner JIANGSU HEVI BIOTECH CO LTD
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