Three-gene co-expression vector for synthesizing tetrahydropyrimidine and application of co-expression vector

A technology of co-expression vector and ectoine, which is applied in the field of enzyme engineering and compound biosynthesis, can solve the problem of low yield and achieve high synthesis efficiency
CN107475282AInactive Publication Date: 2017-12-15HEBEI NORMAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HEBEI NORMAL UNIV
Publication Date
2017-12-15
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses construction of a three-gene co-expression vector for efficiently synthesizing tetrahydropyrimidine and application of the co-expression vector. The construction method comprises the following steps: sequentially inserting coded L-diaminobutyrate aminotransferase (ectA) gene,coded L-diaminobutyric acid acetyltransferase (ectB) gene and coded tetrahydropyrimidine synthase (ectC) gene in pseudo-bacillus firmus into a transformed plasmid pET-22bNS according to an isocaudarner principle, and constructing the three-gene tandem co-expression vector pET-22bNS-EctA / B / C. When the constructed three-gene co-expression vector is transferred into Escherichia coli BL21 (DE3) and is subjected to IPTG induction, after a recombinant strain whole-cell catalysis reaction for 3 hours, the highest yield of tetrahydropyrimidine is 9.8mg / mL, and the synthetic efficiency is 78.4mg / mL / d and is higher than a synthesizing level in similar researches. The three-gene co-expression vector constructed by the invention has the ability of efficiently synthesizing the tetrahydropyrimidine and has an excellent application value.
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Description

technical field

[0001] The invention relates to a construction method and application of a three-gene co-expression vector for synthesizing ectoine, belonging to the technical fields of enzyme engineering and compound biosynthesis. Background technique

[0002] The ectoine compounds are a kind of compatible solutes synthesized by halophilic and halophilic bacteria that can resist external high-salt stress. The ectoine-compatible solutes have a wide range of anti-stress effects, and can stabilize enzyme proteins DNA and cell membrane structure help cells resist various adversities such as freezing, drought, and high-salt hyperosmotic radiation. At present, ectoine compounds have been widely used in industries such as medicine, beauty, fine chemicals and biomanufacturing. The traditional method of synthesizing ectoine is mainly "bacterial milking". This method is to synthesize and accumulate ectoine in a high-salt environment, release ectoine in a low-salt environment, and co...

Claims

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