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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Griffin M, Casadio R, Bergamini CM. Transglutaminases: Nature’s biological glues. Biochem. J. 368: 377–396 (2002)
Yokoyama K, Nio N, Kikuchi Y. Properties and applications of microbial transglutaminase. Appl. Microbiol. Biotechnol. 64: 447–454 (2004)
DeJong G, Koppelman S. Transglutaminase catalyzed reactions: Impact on food applications. J. Food Sci. 67: 2798–2806 (2002)
Marx CK, Hertel TC, Pietzsch M. Purification and activation of a recombinant histidine-tagged pro-transglutaminase after soluble expression in Escherichia coli and partial characterization of the active enzyme. Enzyme Microb. Technol. 42: 568–575 (2008)
Santos M, Torne J. Recent patents on transglutaminase production and applications: A brief review. Recent Patents Biotechnol. 3: 166–174 (2009)
Icekson I, Apelbaum A. Evidence for transglutaminase activity in plant tissue. Plant Physiol. 84: 972–974 (1987)
Margosiak SA, Dharma A, Bruce-Carver MR, Gonzales AP, Louie D, Kuehn GD. Identification of the large subunit of ribulose 1, 5-bisphosphate carboxylase/oxygenase as a substrate for transglutaminase in Medicago sativa L. (Alfalfa). Plant Physiol. 92: 88–96 (1990)
Duca S, Tidu V, Bassi R, Esposito C, Serafmi-Fracassini D. Identification of chlorophyll-a/b proteins as substrates of transglutaminase activity in isolated chloroplasts of Helianthus tuberosus L. Planta 193: 283–289 (1994)
Kang H, Cho YD. Purification and properties of transglutaminase from soybean (Glycine max) leaves. Biochem. Bioph. Res. Co. 223: 288–292 (1996)
Bernet E, Claparols I, Dondini L, Asunción Santos M, Serafini-Fracassini D, Torné JM. Changes in polyamine content, arginine and ornithine decarboxylases and transglutaminase activities during light/dark phases (of initial differentiation) in maize calluses and their chloroplasts. Plant Physiol. Bioch. 37: 899–909 (1999)
Villalobos E, Torné J, Rigau J, Ollés I, Claparols I, Santos M. Immunogold localization of a transglutaminase related to grana development in different maize cell types. Protoplasma. 216: 155–163 (2001)
Villalobos E, Santos M, Talavera D, Rodrlguez-Falcón M, Torné J. Molecular cloning and characterization of a maize transglutaminase complementary DNA. Gene 336: 93–104 (2004)
Carvajal P, Villalobos E, Campos A, Torné JM, Barberà E, Santos M. Zea mays L. transglutaminase expression in Escherichia coli. Microb. Cell. Fact. 5: P8 (2006)
Carvajal P, Gibert J, Campos N, Lopera O, Barberà E, Torné JM, Santos M. Activity of maize transglutaminase overexpressed in Escherichia coli inclusion bodies: An alternative to protein refolding. Biotechnol. Prog. 27: 232–240 (2011)
Daly R, Hearn MTW. Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production. J. Mol. Recognit. 18: 119–138 (2005)
Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev. 24: 45–66 (2000)
Zhang J, Wang X, Su E, Fang G, Ren Y, Wei D. A new fermentation strategy for S-adenosylmethionine production in recombinant Pichia pastoris. Biochem. Eng. J. 41: 74–78 (2008)
Kang HK, Yun SI, Lim TY, Xia YM, Kim D. Cloning of levansucrase from Leuconostoc mesenteroides and its expression in Pichia pastoris. Food Sci. Biotechnol. 20: 277–281 (2011)
Ryu HJ, Seo ES, Kang HK, Kim YM, Kim D. Expression, purification, and characterization of human intestinal maltase secreted from Pichia pastoris. Food Sci. Biotechnol. 20: 561–565 (2011)
Zhang L, Zhang L, Yi H, Du M, Ma C, Han X, Feng Z, Jiao Y, Zhang Y. Enzymatic characterization of transglutaminase from Streptomyces mobaraensis DSM 40587 in high salt and effect of enzymatic cross-linking of yak milk proteins on functional properties of stirred yogurt. J. Dairy Sci. 95: 3559–3568 (2012)
Qin LX, Wang L, Zhang LW. Cloning and expression of maize transglutaminase in Escherichia coli. Food Sci. 31: 177–181 (2010)
Chen CC, Wu PH, Huang CT, Cheng KJ. A Pichia pastoris fermentation strategy for enhancing the heterologous expression of an Escherichia coli phytase. Enzyme Microb. Technol. 35: 315–320 (2004)
Grossowicz N, Wainfan E, Borek E, Waelsch H. The enzymatic formation of hydroxamic acids from glutamine and asparagine. J. Biol. Chem. 187: 111–125 (1950)
Wang H, Wang Q, Zhang F, Huang Y, Ji Y, Hou Y. Protein expression and purification of human Zbtb7A in Pichia pastoris via gene codon optimization and synthesis. Protein Expr. Purif. 60: 97–102 (2008)
Hermanrud CE, Lucas CL, Sykes M, Huang CA, Wang Z. Expression and purification of soluble murine CD40L monomers and polymers in yeast Pichia pastoris. Protein Expr. Purif. 76: 115–120 (2011)
Kupcsulik B, Sevella B. Effect of methanol concentration on the recombinant Pichia pastoris Muts fermentation. Chem. Eng. 48: 73–87 (2004)
Hu XQ, Chu J, Zhang SL, Zhuang YP, Wang YH, Zhu S, Zhu ZG, Yuan ZY. A novel feeding strategy during the production phase for enhancing the enzymatic synthesis of S-adenosyl-l-methionine by methylotrophic Pichia pastoris. Enzyme Microb. Technol. 40: 669–674 (2007)
Smiddy M, Martin JEGH, Kelly A, De Kruif C, Huppertz T. Stability of casein micelles cross-linked by transglutaminase. J. Dairy Sci. 89: 1906–1914 (2006)
Moon JH, Hong YH, Huppertz T, Fox PF, Kelly AL. Properties of casein micelles cross-linked by transglutaminase. Int. J. Dairy Technol. 62: 27–32 (2009)
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, H., Cui, Y., Zhang, L. et al. Heterologous expression and purification of Zea mays transglutaminase in Pichia pastoris . Food Sci Biotechnol 23, 1507–1513 (2014). https://doi.org/10.1007/s10068-014-0206-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10068-014-0206-1