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

Yurimoto, Hiroya; Yamane, Maiko; Kikuchi, Yoshimi; Matsui, Hiroshi; Kato, Nobuo; Sakai, Yasuyoshi

doi:10.1271/bbb.68.2058

Cited by 45 publications

(55 citation statements)

References 24 publications

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“…These vectors were transfected into either COS-1 cells or Pichia pastoris strain SMD1163 (his4 pep4 prb1), 15 respectively. For Pichia expression, the DNA coding region for TME5 was synthesized according to the preferred codon usage of the methylotrophic yeasts 16 and the transformant cells were grown to highcell density on methanol medium. 16 The generated TME5 proteins were purified with a His-tagged protein purification kit (MBL, Aichi, Japan) or Ni Sepharose 6 Fast Flow (GE Healthcare, Little Chalfont, UK).…”

Section: Generation Of Tme5mentioning

confidence: 99%

The fifth epidermal growth factor-like region of thrombomodulin exerts cytoprotective function and prevents SOS in a murine model

Ikezoe

Yang

Nishioka

et al. 2016

Bone Marrow Transplant

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The present study found that the fifth epidermal growth factor-like domain of thrombomodulin (TME5) possesses the cytoprotective function in association with an increase in levels of anti-apoptotic myeloid cell leukemia-1 protein in an activated protein C-independent manner in human umbilical vein endothelial cells (HUVECs). Importantly, TME5 counteracted calcineurin inhibitor-induced vascular permeability and successfully prevented monocrotaline-induced sinusoidal obstruction syndrome (SOS) in a murine model. Taken together, TME5 may be useful for preventing or treating lethal complications that develop after hematopoietic stem cell transplantation such as SOS and thrombotic microangiopathy in which endothelial cell damage has a role.

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Section: Generation Of Tme5mentioning

confidence: 99%

The fifth epidermal growth factor-like region of thrombomodulin exerts cytoprotective function and prevents SOS in a murine model

Ikezoe

Yang

Nishioka

et al. 2016

Bone Marrow Transplant

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“…It has been reported that mature MTG from S. mobaraensis is directly obtained in Corynebacterium glutamicum by the coexpression of pro-MTG and a protease (35,36). In addition, Yurimoto et al revealed that the coexpression of proregion and MTG from S. mobaraensis in methylotrophic yeasts could also secrete mature MTG (37). However, these expression systems have the drawbacks of long culture times and high cultivation costs.…”

Section: Sdb-mtg(⌬d1)mentioning

confidence: 99%

pH-Dependent Activation of Streptomyces hygroscopicus Transglutaminase Mediated by Intein

Liu

Cui

et al. 2014

Appl Environ Microbiol

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bMicrobial transglutaminase (MTG) from Streptomyces is naturally secreted as a zymogen (pro-MTG), which is then activated by the removal of its N-terminal proregion by additional proteases. Inteins are protein-intervening sequences that catalyze protein splicing without cofactors. In this study, a pH-dependent Synechocystis sp. strain PCC6803 DnaB mini-intein (SDB) was introduced into pro-MTG to simplify its activation process by controlling pH. The recombinant protein (pro-SDB-MTG) was obtained, and the activation process was determined to take 24 h at pH 7 in vitro. To investigate the effect of the first residue in MTG on the activity and the cleavage time, two variants, pro-SDB-MTG(D1S) and pro-SDB-MTG(⌬D1), were expressed, and the activation time was found to be 6 h and 30 h, respectively. The enzymatic property and secondary structure of the recombinant MTG and two variants were similar to those of the wild type, indicating that the insertion of mini-intein did not affect the function of MTG. This insignificant effect was further illustrated by molecular dynamics simulations. This study revealed a controllable and effective strategy to regulate the activation process of pro-MTG mediated by a mini-intein, and it may have great potential for industrial MTG production.

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“…[39][40][41] An efficient secretion system for the production of active transglutaminase was also developed. 42) 1. Oxidase production within peroxisomes of the aod1Á strain From a biotechnological point of view, peroxisomes of the methylotrophic yeasts are attractive organelles in which to pack the produced heterologous proteins where they are protected from degradation by cytosolic proteases and, in the case of proteins of a toxic nature, have to remain separated from the cytosol.…”

Section: Production Of Useful Enzymes In C Boidiniimentioning

confidence: 99%

“…We have reported the secretion of TGase in an enzymatically active form by the methylotrophic yeast as an expression host. 42) The coding regions for the pro-peptide and the mature TGase were synthesized according to the preferred codon usage in C. boidinii. Then we constructed different kinds of expression cassettes under the control of the AOD1 promoter (Fig.…”

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

Cited by 45 publications

References 24 publications

The fifth epidermal growth factor-like region of thrombomodulin exerts cytoprotective function and prevents SOS in a murine model

The fifth epidermal growth factor-like region of thrombomodulin exerts cytoprotective function and prevents SOS in a murine model

pH-Dependent Activation of Streptomyces hygroscopicus Transglutaminase Mediated by Intein

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

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