Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris

Hongbo Li; Yanhua Cui; Lanwei Zhang; Huaxi Yi; Xue Han; Yuehua Jiao; Ming Du; Rongbo Fan; Shuang Zhang

doi:10.1007/s10068-014-0206-1

Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris

Hongbo Li
, Yanhua Cui
, Lanwei Zhang^*
, Huaxi Yi
, Xue Han
, Yuehua Jiao
, Ming Du
, Rongbo Fan
, Shuang Zhang

^*Corresponding author for this work

Harbin Institute of Technology
Qingdao Agricultural University

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Transglutaminases (TGases) are a family of enzymes that catalyze the cross-linking of proteins and are widely used in the food industry to improve the texture of dairy, meat, and bread products. Zea mays transglutaminase (TGZ) is a new type of TGase with a wide potential. TGZ was expressed in the yeast Pichia pastoris under an alcohol oxidase promoter. Maximal expression of recombinant TGZ was achieved by inducing recombinant GS115 (pPIC9K-tgz) in BMMY medium using 1.5% methanol for 96 h. Secreted TGZ was initially separated using Superdex 200 resin and further purified on cation exchange resin. The activity of TGZ following purification was 0.32 U/mg of protein. The polymerization effect of TGZ on casein catalyzed by recombinant TGZ was slightly lower than the effect of microbial transglutaminase (MTG). TGZ is a new potential additive for the food industry.

Original language	English
Pages (from-to)	1507-1513
Number of pages	7
Journal	Food Science and Biotechnology
Volume	23
Issue number	5
DOIs	https://doi.org/10.1007/s10068-014-0206-1
State	Published - 14 Oct 2014

Keywords

Pichia pastoris GS115
Zea mays transglutaminase
polymerization effect
protein expression
purification

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10.1007/s10068-014-0206-1

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title = "Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris",

abstract = "Transglutaminases (TGases) are a family of enzymes that catalyze the cross-linking of proteins and are widely used in the food industry to improve the texture of dairy, meat, and bread products. Zea mays transglutaminase (TGZ) is a new type of TGase with a wide potential. TGZ was expressed in the yeast Pichia pastoris under an alcohol oxidase promoter. Maximal expression of recombinant TGZ was achieved by inducing recombinant GS115 (pPIC9K-tgz) in BMMY medium using 1.5\% methanol for 96 h. Secreted TGZ was initially separated using Superdex 200 resin and further purified on cation exchange resin. The activity of TGZ following purification was 0.32 U/mg of protein. The polymerization effect of TGZ on casein catalyzed by recombinant TGZ was slightly lower than the effect of microbial transglutaminase (MTG). TGZ is a new potential additive for the food industry.",

keywords = "Pichia pastoris GS115, Zea mays transglutaminase, polymerization effect, protein expression, purification",

author = "Hongbo Li and Yanhua Cui and Lanwei Zhang and Huaxi Yi and Xue Han and Yuehua Jiao and Ming Du and Rongbo Fan and Shuang Zhang",

note = "Publisher Copyright: {\textcopyright} 2014, The Korean Society of Food Science and Technology and Springer Science+Business Media Dordrecht.",

year = "2014",

month = oct,

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doi = "10.1007/s10068-014-0206-1",

language = "英语",

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TY - JOUR

T1 - Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris

AU - Li, Hongbo

AU - Cui, Yanhua

AU - Zhang, Lanwei

AU - Yi, Huaxi

AU - Han, Xue

AU - Jiao, Yuehua

AU - Du, Ming

AU - Fan, Rongbo

AU - Zhang, Shuang

PY - 2014/10/14

Y1 - 2014/10/14

N2 - Transglutaminases (TGases) are a family of enzymes that catalyze the cross-linking of proteins and are widely used in the food industry to improve the texture of dairy, meat, and bread products. Zea mays transglutaminase (TGZ) is a new type of TGase with a wide potential. TGZ was expressed in the yeast Pichia pastoris under an alcohol oxidase promoter. Maximal expression of recombinant TGZ was achieved by inducing recombinant GS115 (pPIC9K-tgz) in BMMY medium using 1.5% methanol for 96 h. Secreted TGZ was initially separated using Superdex 200 resin and further purified on cation exchange resin. The activity of TGZ following purification was 0.32 U/mg of protein. The polymerization effect of TGZ on casein catalyzed by recombinant TGZ was slightly lower than the effect of microbial transglutaminase (MTG). TGZ is a new potential additive for the food industry.

AB - Transglutaminases (TGases) are a family of enzymes that catalyze the cross-linking of proteins and are widely used in the food industry to improve the texture of dairy, meat, and bread products. Zea mays transglutaminase (TGZ) is a new type of TGase with a wide potential. TGZ was expressed in the yeast Pichia pastoris under an alcohol oxidase promoter. Maximal expression of recombinant TGZ was achieved by inducing recombinant GS115 (pPIC9K-tgz) in BMMY medium using 1.5% methanol for 96 h. Secreted TGZ was initially separated using Superdex 200 resin and further purified on cation exchange resin. The activity of TGZ following purification was 0.32 U/mg of protein. The polymerization effect of TGZ on casein catalyzed by recombinant TGZ was slightly lower than the effect of microbial transglutaminase (MTG). TGZ is a new potential additive for the food industry.

KW - Pichia pastoris GS115

KW - Zea mays transglutaminase

KW - polymerization effect

KW - protein expression

KW - purification

UR - https://www.scopus.com/pages/publications/84918516204

U2 - 10.1007/s10068-014-0206-1

DO - 10.1007/s10068-014-0206-1

M3 - 文章

AN - SCOPUS:84918516204

SN - 1226-7708

VL - 23

SP - 1507

EP - 1513

JO - Food Science and Biotechnology

JF - Food Science and Biotechnology

IS - 5

ER -

Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris

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Keywords

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