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Recombinant strain of Hansenula polymorpha and its application in glycerol biosynthesis

A technology of Hansenula polymorpha and recombinant strains, applied in the direction of microorganism-based methods, microorganisms, microorganisms, etc., can solve the problems of glycerol production and substrate utilization to be improved, and achieve high-density fermentation, low cost, and effective The effect of biosynthesis

Inactive Publication Date: 2019-08-27
NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION +1
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Problems solved by technology

[0005] Zhao Dezhi et al. used the method of low-energy ion implantation to mediate the total DNA of yeast genome to transform yeast, and obtained a strain of yeast engineering strain with high glycerol production. However, the yield of glycerol and substrate utilization still need to be improved.

Method used

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Embodiment Construction

[0020] The present invention is described in detail below in conjunction with embodiment.

[0021] The present invention uses Hansenula polymorpha (H.polymorpha) DL-1 as the starting strain (the source is ATCC No.26012), and uses low-energy ion implantation to mediate random transformation of Hansenula polymorpha (H.polymorpha) DL- 1. Use the high-salt solid medium selective screening method to conduct preliminary screening of recombinant strains; carry out fermentation and culture of the preliminary screened recombinant strains, and use paper chromatography and gas chromatography to measure the yield of glycerol in the fermentation broth, thereby screening to obtain a A recombinant strain of Hansenula polymorpha that can biosynthesize glycerol more efficiently, and its genetic stability was analyzed.

[0022] (1) culture medium

[0023] The composition of the high-salt solid medium is: glucose 20.0g / L, peptone 20.0g / L, yeast extract 10.0g / L, agar 20.0g / L, sodium chloride 180...

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Abstract

The present invention provides a Hansenula polymorpha recombinant strain and its application in glycerol biosynthesis, mainly using low-energy ion implantation technology to mediate the method of transforming yeast genome total DNA into yeast to construct a high-salt-tolerant glycerol high-yielding recombinant strain . The recombinant bacterial strain provided by the invention produces glycerol through biotransformation by feeding batches of substrates, and the yield of glycerol and the conversion rate of substrate glucose can respectively reach 97.29g / L and 83.27% during fermentation.

Description

technical field [0001] The invention belongs to the field of ion beam-mediated transgenes, and in particular relates to the construction of high-salt-resistant recombinant yeast strains by using low-energy ion implantation to mediate the transformation of yeast genome total DNA and its application in glycerol biosynthesis. Background technique [0002] Glycerin (Glycerin), also known as glycerol, trihydroxypropane, molecular formula C 3 h 8 o 3 , relative density 1.26, melting point 17.8°C, boiling point 290.0°C. Glycerin is a colorless or yellowish transparent viscous liquid, non-toxic, odorless, non-corrosive, and has strong hygroscopicity. As an important organic chemical raw material, glycerin is widely used in military industry, food, medicine, daily chemical industry and other industries. my country's glycerin production capacity is about 86,000-90,000 tons, of which the production capacity of saponification method is 60,000 tons, the production capacity of chemica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12P7/20C12R1/78
CPCC12P7/20C12N1/165C12R2001/78
Inventor 李永东钱卫东倪红霞刘昱慧
Owner NINGBO MUNICIPAL CENT FOR DISEASE CONTROL & PREVENTION
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