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Preparation and quick screening method of yeast fusant

A technology for yeast fusogen and screening method, which is applied in the field of yeast fusant preparation and high-throughput rapid screening, can solve the problems such as reports on the rapid screening method of Schizosaccharomyces pombe and Hansenula polymorpha that have not yet been seen. , to achieve the effect of efficient production and fast growth

Active Publication Date: 2015-11-18
SHAANXI UNIV OF SCI & TECH
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Problems solved by technology

[0004] At present, there are no reports on the preparation of protoplast fusions of Schizosaccharomyces pombe and Hansenula polymorpha and rapid screening methods

Method used

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Embodiment Construction

[0030] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0031] The strains involved in the present invention: Hansenula polymorpha (Hansenulapolymorpha) DL-1, derived from the China Industrial Microorganism Collection Center, with a preservation number of ATCC No.26012; Schizosaccharomycespombe (S.promb2.1794), purchased from the Institute of Microbiology, Chinese Academy of Sciences.

[0032] The culture medium that the present invention relates to:

[0033] 1) YPD medium: Yeast extract powder 10g / L, peptone 20g / L, glucose 20g / L, natural pH, (additional agar powder 20g / L is required to prepare solid medium).

[0034] 2) KCl hyperosmotic medium: yeast extract powder 10g / L, peptone 20g / L, glucose 20g / L, agar powder 20g / L, KCl 40g / L, pH natural.

[0035] 3) Vitamin K 3 Selection medium: add a certain amount of vitamin K to YPD medium 3 .

[0036] 4) Wort medium: Yeast extract powder 3g / L, peptone 4g / L, ...

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Abstract

The invention provides a preparation and quick screening method of yeast fusant. In order to realize mass production of coenzyme Q10, a method of protoplast fusion is utilized to fuse schizosaccharomyces pombe and hensenula polymorpha and combined with a method for efficiently screening target yeast fusant to quickly and efficiently obtain target yeast fusant having excellent characteristics of two yeast strains, and the target yeast fusant has the advantages of high growing speed, resistance to high temperature and capability of efficiently producing the coenzyme Q10. According to the yeast fusant screening method, growing performance including growing speed, resistance to high temperature and target metabolite of thalli is taken as a screening marker; specifically, temperature tolerance and a culture medium containing vitamin K3 which is a structural analog of the coenzyme Q10 are taken as screening strategies, and the target fusant can be directly screened in a high-throughput manner. The preparation and quick screening method is a yeast improving method which is simple, quick, efficient, economical, practical and safe.

Description

technical field [0001] The invention relates to the preparation of yeast fusion son and its high-throughput rapid screening method. Background technique [0002] coenzyme Q 10 As an important electronic hydrogen carrier in the respiratory chain, it can reversibly carry out oxidation-reduction reactions, and widely exists in humans, animals, plants and most microorganisms. At present, coenzyme Q is produced by microbial fermentation 10 The main strain of S. pombe is Schizosaccharomycespombe (S.sprombe), but S. pombe grows slowly and requires strict growth conditions, which seriously affects the coenzyme Q 10 production efficiency. In recent years, researchers have further genetically modified it through ultraviolet mutagenesis, ion beam mutagenesis, etc., making it a coenzyme Q 10 production carrier. [0003] As food-grade Hansenula polymorpha (H.polymorpha), it has attracted much attention in recent years because of its high safety and obvious advantages. Its advantage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/04C12N1/19C12R1/78C12R1/645
Inventor 钱卫东周颖欣吴启航宁肖肖
Owner SHAANXI UNIV OF SCI & TECH
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