Chemical Synthesis of the Gene for Microbial Transglutaminase from<i>Streptoverticillium</i>and Its Expression in<i>Escherichia coli</i>

Takehana, Shino; Washizu, Kinya; Ando, Kentaro; Koikeda, Satoshi; Takeuchi, Kikuko; Matsui, Hiroshi; Motoki, Masao; Takagi, Hiroshi

doi:10.1271/bbb.58.88

Cited by 51 publications

(36 citation statements)

References 3 publications

(3 reference statements)

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“…Induction of TGase in transformed Escherichia coli containing a synthetic gene coding for the mature enzyme results in inhibition of cell growth followed by lysis. Cell death is obviously caused by the TGase-catalysed crosslinking of essential cytosolic proteins [13].…”

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confidence: 99%

Bacterial pro‐transglutaminase from Streptoverticillium mobaraense

Pasternack

Dorsch

Otterbach

et al. 1998

European Journal of Biochemistry

142

117

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The zymogen of bacterial transglutaminase was found during cultivation of Streptoverticillium mobaraense (DSMZ strain) using rabbit antibodies raised against the active enzyme. Ion‐exchange chromatography at pH 5.0 yielded a highly purified pro‐enzyme. Structure information was obtained by means of Edman degradation and analysis of PCR amplified nucleotide fragments. The data revealed an excess of negatively charged amino acids in the pro‐region resulting in a decreased isoelectric point of the zymogen. Additionally, the new sequence gave rise to some modifications to the previously published hypothetical structure of prepro‐transglutaminase derived from genomic DNA [Washizu, K., Ando, K., Koikeda, S., Hirose, S., Matsuura, A., Takagi, H., Motoki, M. & Takeuchi, K. (1994) Biosci. Biotechnol. Biochem. 58, 82−87]. Inactive transglutaminase, which carries an activation peptide of 45 amino acids, has a calculated molecular mass of 42 445 Da. Its pro‐region provides for both suppression of activity and increased thermostability. Furthermore, it could be shown that the micro‐organism produces a protease which cleaves pro‐transglutaminase at the C‐side of Pro45. Rapid transformation of the mature enzyme also occurs by addition of other proteases. During conversion, 43 and 41 amino acid peptides are released by bovine trypsin and dispase from Bacillus polymyxa, respectively. The detection of endogenous substrates in the murein layer makes discussion of the physiological role of bacterial transglutaminases necessary.

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mentioning

confidence: 99%

Bacterial pro‐transglutaminase from Streptoverticillium mobaraense

Pasternack

Dorsch

Otterbach

et al. 1998

European Journal of Biochemistry

142

117

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“…The DNA coding regions for pro and mature TGase were chemically synthesized according to the preferred codon usage of the two methylotrophic yeast strains, and the sequence of this synthesized fragment on pOMPA BTG+3 12) was confirmed by DNA-sequencing and submitted to DDBJ (AB181959).…”

Section: Methodsmentioning

confidence: 99%

“…The pro-peptide appears to be cleaved by specific metalloprotease in the culture medium of S. mobaraensis. 10,11) Heterologous gene expression of microbial TGase has also been reported using Escherichia coli as the host, 12) although difficulties were encountered in obtaining secretion of TGase in an active form. In vitro refolding of the intracellularly produced mature TGase was found to be necessary to obtain an active enzyme.…”

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confidence: 99%

The Pro-peptide ofStreptomyces mobaraensisTransglutaminase Functions incisand intransto Mediate Efficient Secretion of Active Enzyme from Methylotrophic Yeasts

Yurimoto

Yamane

Kikuchi

et al. 2004

Bioscience, Biotechnology, and Biochemistry

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“…50,51) Heterologous gene expression of microbial TGase has also been reported using Escherichia coli as the host, although difficulties were encountered in getting TGase secreted in an active form. 52) Recently, secretion of active TGase was reported by coexpression of a subtilisin-like protease in a Streptomyces or Corynebacterium expression system. 48) But co-expression of the protease was difficult to control, and protease overproduction may lead to degradation of the produced TGase.…”

Section: Efficient Secretion Of Transglutaminasementioning

confidence: 99%

Molecular Basis of Methanol-Inducible Gene Expression and Its Application in the Methylotrophic YeastCandida boidinii

Yurimoto

2009

Bioscience, Biotechnology, and Biochemistry

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Methanol is a promising feedstock for biotechnological and chemical processes as well as a primary source of energy to replace coal and petroleum. Methylotrophic yeasts, that can utilize methanol as the sole source of carbon and energy, have been studied intensively in terms of both physiological activities and potential applications. During growth on methanol, the enzymes involved in methanol metabolism are massively produced in these yeasts, indicating that the gene promoters of these enzymes are strong methanol-inducible promoters. Using these promoters, high-level heterologous gene expression systems have been developed in several methylotrophic yeast strains, such as Pichia pastoris, Hansenula polymorpha, and Candida boidinii. To achieve efficient industrial use of methanol and efficient protein production by methylotrophic yeasts, it is important to elucidate the molecular basis of methanolinducible gene expression in these yeasts. This review describes recent advances in understanding of the regulation of methanol-inducible gene expression and the molecular mechanism of transcriptional activation in the methylotrophic yeast C. boidinii. Application of this gained knowledge led to successful production of useful enzymes in this yeast, which is also reviewed.

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Chemical Synthesis of the Gene for Microbial Transglutaminase fromStreptoverticilliumand Its Expression inEscherichia coli

Cited by 51 publications

References 3 publications

Bacterial pro‐transglutaminase from Streptoverticillium mobaraense

Bacterial pro‐transglutaminase from Streptoverticillium mobaraense

The Pro-peptide ofStreptomyces mobaraensisTransglutaminase Functions incisand intransto Mediate Efficient Secretion of Active Enzyme from Methylotrophic Yeasts

Molecular Basis of Methanol-Inducible Gene Expression and Its Application in the Methylotrophic YeastCandida boidinii

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