High-level expression of the chemically synthesized gene for microbial transglutaminase from Streptoverticillium in Escherichia coli

Bioscience, Biotechnology, and Biochemistry
M KawaiH Takagi

Abstract

We developed a novel approach for the high-level production of a microbial transglutaminase (TGase) from Streptoverticillium in E. coli. The direct expression of the TGase gene in E. coli cells did not cause overproduction, probably due to the harmful influence of TGase activity, which introduces covalent crosslinks between proteins. Therefore, we fused the chemically synthesized TGase gene coding for the entire 331 amino acid residues at the amino terminus to a bacteriophage T7 gene 10 leader peptide (260 amino acids) using an inducible expression vector. The TGase gene was expressed as inclusion bodies in the E. coli cytoplasm. Restoring 15 amino acid residues upstream of the amino terminus of the mature TGase by a two-step deletion of the fusion sequence facilitated solubilization and subsequent proteolytic cleavage, thus releasing mature TGase. Although the mature form had less TGase activity than native TGase, because of the poor refolding rate, these results suggest that this system is suitable for the efficient production of TGase.

Citations

Jan 29, 2002·Analytical Biochemistry·Nobuhisa ShimbaEi-ichiro Suzuki
Oct 21, 2015·Protein Science : a Publication of the Protein Society·Mathias RickertArvind Rajpal
Dec 21, 2010·Protein Expression and Purification·Christian SommerMarkus Pietzsch
Aug 19, 2008·Trends in Biotechnology·Yang Zhu, Johannes Tramper
Aug 14, 2012·Biotechnology and Bioengineering·Jae-Hun LeeByung-Gee Kim
Jan 1, 2010·Biotechnology & Genetic Engineering Reviews·Dongxu ZhangJian Chen
Jan 4, 2020·World Journal of Microbiology & Biotechnology·Lovaine DuarteMarco Antônio Záchia Ayub

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