Publication: A Novel Potential Signal Peptide Sequence and Overexpression of ER-Resident Chaperones Enhance Heterologous Protein Secretion in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica
Issued Date
2016-02-01
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ISSN
15590291
02732289
02732289
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2-s2.0-84959464555
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Mahidol University
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SCOPUS
Bibliographic Citation
Applied Biochemistry and Biotechnology. Vol.178, No.4 (2016), 710-724
Suggested Citation
Niran Roongsawang, Aekkachai Puseenam, Supattra Kitikhun, Kittapong Sae-Tang, Piyanun Harnpicharnchai, Takao Ohashi, Kazuhito Fujiyama, Witoon Tirasophon, Sutipa Tanapongpipat A Novel Potential Signal Peptide Sequence and Overexpression of ER-Resident Chaperones Enhance Heterologous Protein Secretion in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica. Applied Biochemistry and Biotechnology. Vol.178, No.4 (2016), 710-724. doi:10.1007/s12010-015-1904-8 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/43155
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Title
A Novel Potential Signal Peptide Sequence and Overexpression of ER-Resident Chaperones Enhance Heterologous Protein Secretion in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica
Abstract
© 2015, Springer Science+Business Media New York. The thermotolerant methylotrophic yeast Ogataea thermomethanolica is a host for heterologous protein expression via secretion to the culture medium. Efficient secretion is a major bottleneck for heterologous protein production in this strain. To improve protein secretion, we explored whether the use of a native signal peptide sequence for directing heterologous protein secretion and overexpression of native ER-resident chaperone genes could improve heterologous protein secretion in O. thermomethanolica. We cloned and characterized genes encoding α-mating factor (Otα-MF) and ER-resident chaperones OtBiP, OtCNE1, and OtPDI. The pre and pre-pro sequences of Otα-MF were shown to promote higher secretion of heterologous endoxylanase comparing with the classical pre-pro sequence of Saccharomyces cerevisiae. However, in the case of heterologous glycosylated phytase, only the Otα-MF pre-pro sequence significantly enhanced protein secretion. The effect of chaperone overexpression on heterologous protein secretion was tested in cotransformant cells of O. thermomethanolica. Overexpression of ER-resident chaperones improved protein secretion depending on heterologous protein. Overexpression of OtBiP, OtCNE1, and OtPDI significantly increased unglycosylated endoxylanase secretion at both 30 and 37 °C while only OtBiP overexpression enhanced glycosylated phytase secretion at 30 °C. These observations suggested the possibility to improve heterologous protein secretion in O. thermomethanolica.
