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Heterologous protein expression in Pichia thermomethanolica BCC16875, a thermotolerant methylotrophic yeast and characterization of N-linked glycosylation in secreted protein

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논문

Heterologous protein expression in Pichia thermomethanolica BCC16875, a thermotolerant methylotrophic yeast and characterization of N-linked glycosylation in secreted protein

학술지

FEMS microbiology letters

저자명

Tanapongpipat, Sutipa; Promdonkoy, Peerada; Watanabe, Toru; Tirasophon, Witoon; Roongsawang, Niran; Chiba, Yasunori; Eurwilaichitr, Lily

초록

<P><B>Abstract</B></P><P>This study describes <I>Pichia thermomethanolica </I>BCC16875, a new methylotrophic yeast host for heterologous expression. Both methanol&#8208;inducible alcohol oxidase (<I>AOX1</I>) and constitutive glyceraldehyde&#8208;3&#8208;phosphate dehydrogenase (<I>GAP</I>) promoters from <I>Pichia pastoris</I> were shown to drive efficient gene expression in this host. Recombinant phytase and xylanase were expressed from both promoters as secreted proteins, with the former showing different patterns of <I>N</I>&#8208;glycosylation dependent on the promoter used and culture medium. In addition, growth temperature also had an effect on <I>N</I>&#8208;glycan modification of cell wall mannoproteins. The major glycoprotein oligosaccharide species produced from <I>P.?thermomethanolica </I>BCC16875 is Man<SUB>8&#8208;12</SUB>GlcNAc<SUB>2</SUB>, which is similar to that from other methylotrophs. Moreover, mannosylphosphate and &alpha;&#8208;1,6&#8208; and &alpha;&#8208;1,2&#8208;linked mannose modifications of heterologous secreted protein were also detected. The attainably high level of protein production in complement to distinctive thermotolerance rarely found in other industrial yeasts makes this microorganism an attractive host for large&#8208;scale fermentation.</P>

발행연도

2012

ISSN

0378-1097

ISSN

1574-6968

334

2

페이지

pp.127-134

주제어

N&#x2010; linked glycans; Pichia thermomethanolica; thermotolerant;

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1 2023-12-11
2 2023-12-11

논문; 2012-07-18

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