Recombinant pichia pastoris strain regulating expression of glutamine transaminase in Zea mays by using FLD1 promoter and construction method thereof

A kind of Pichia pastoris, glutamine technology

Inactive Publication Date: 2019-06-25
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
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[0005] Through the search, no patent publications related to

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  • Recombinant pichia pastoris strain regulating expression of glutamine transaminase in Zea mays by using FLD1 promoter and construction method thereof
  • Recombinant pichia pastoris strain regulating expression of glutamine transaminase in Zea mays by using FLD1 promoter and construction method thereof
  • Recombinant pichia pastoris strain regulating expression of glutamine transaminase in Zea mays by using FLD1 promoter and construction method thereof

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[0026] The present invention will be described in further detail below in conjunction with specific examples. The following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention.

[0027] The raw materials used in the present invention, unless otherwise specified, are conventional commercially available products; the methods used in the present invention, unless otherwise specified, are conventional methods in the art.

[0028] The medium used in the present invention can be as follows:

[0029] LB medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L.

[0030] YPD medium: peptone 20g / L, yeast extract 10g / L, glucose 20g / L.

[0031] The solid medium is agar added to the liquid medium in an amount of 20 g / L.

[0032] Fermentation medium: peptone 20g / L, yeast extract 10g / L, amino acid-free yeast nitrogen source 13.4g / L, sorbitol 5.4g / L, biotin 0.4mg / L, 100mL 1mol / L (pH 6.0) phosphoric acid salt buffer.

[00...

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Abstract

The invention relates to a recombinant pichia pastoris strain regulating expression of glutamine transaminase in Zea mays by using a FLD1 promoter and a construction method thereof. The recombinant pichia pastoris strain is a genetically engineered strain obtained by transferring a transglutaminase gene derived from Zea mays into pichia pastoris, the recombinant pichia pastoris strain is obtainedby replacing an alcohol oxidase promoter PAOX1 in the pPIC9K plasmids to a formaldehyde dehydrogenase promoter PFLD1, and connecting with the glutamine transaminase gene into a loop to form a recombinant expression vector. The recombinant strain is easy to culture has strong secreting capability, is beneficial to the large expression of glutamine transaminase, has the characteristics of safety andreliability, and has important application prospects.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a recombinant Pichia pastoris strain and a construction method using the FLD1 promoter to regulate the expression of glutamine transaminase in corn, more specifically, the construction of the recombinant plasmid pFLD9k-tgz and its activation Sub P FLD1 Regulating the expression of sticky maize transglutaminase in Pichia pastoris. Background technique [0002] Transglutaminase (Transglutaminase, TGase, EC 2.3.2.13) is a kind of acyl transfer reaction between glutamine residues and lysine residues to form lysine isopeptide bonds, so as to achieve protein intramolecular , Intermolecular cross-linking enzymes. TGase can improve the gelation, solubility, emulsification, foaming and water holding capacity of food without changing the flavor and color of the product. At the same time, this cross-linking reaction can transfer essential amino acids to proteins, thereby improv...

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Application Information

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IPC IPC(8): C12N1/19C12N15/81C12N15/66C12N9/10C12R1/84
Inventor 李洪波李春爽郑姗韩翼宇
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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