Methanol-inducible gene expression and heterologous protein production in the methylotrophic yeast Candida boidinii

2009 ◽  
Vol 53 (2) ◽  
pp. 85 ◽  
Author(s):  
Hiroya Yurimoto ◽  
Yasuyoshi Sakai
Keyword(s):  
Gene Expression ◽  
Protein Production ◽  
Heterologous Protein ◽  
Methylotrophic Yeast ◽  
Heterologous Protein Production ◽  
Candida Boidinii ◽  
Inducible Gene Expression ◽  
Inducible Gene

scholarly journals Molecular Characterization of Hap Complex Components Responsible for Methanol-Inducible Gene Expression in the Methylotrophic Yeast Candida boidinii

2015 ◽  
Vol 14 (3) ◽  
pp. 278-285 ◽  
Author(s):  
Saori Oda ◽  
Hiroya Yurimoto ◽  
Nobuhisa Nitta ◽  
Yu Sasano ◽  
Yasuyoshi Sakai
Keyword(s):  
Gene Expression ◽  
Carbon Sources ◽  
Methylotrophic Yeast ◽  
Candida Boidinii ◽  
Promoter Regions ◽  
Inducible Gene Expression ◽  
Content Type ◽  
Inducible Promoters ◽  
Hap Complex ◽  
Inducible Gene

ABSTRACT We identified genes encoding components of the Hap complex, CbHAP2 , CbHAP3 , and CbHAP5 , as transcription factors regulating methanol-inducible gene expression in the methylotrophic yeast Candida boidinii . We found that the Cbhap2 Δ, Cbhap3 Δ, and Cbhap5 Δ gene-disrupted strains showed severe growth defects on methanol but not on glucose and nonfermentable carbon sources such as ethanol and glycerol. In these disruptants, the transcriptional activities of methanol-inducible promoters were significantly decreased compared to those of the wild-type strain, indicating that CbHap2p, CbHap3p, and CbHap5p play indispensable roles in methanol-inducible gene expression. Further molecular and biochemical analyses demonstrated that CbHap2p, CbHap3p, and CbHap5p localized to the nucleus and bound to the promoter regions of methanol-inducible genes regardless of the carbon source, and heterotrimer formation was suggested to be necessary for binding to DNA. Unexpectedly, distinct from Saccharomyces cerevisiae , the Hap complex functioned in methanol-specific induction rather than glucose derepression in C. boidinii . Our results shed light on a novel function of the Hap complex in methanol-inducible gene expression in methylotrophic yeasts.

scholarly journals Multiplexed CRISPR-mediated engineering of protein secretory pathway genes in the thermotolerant methylotrophic yeast Ogataea thermomethanolica

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261754
Author(s):  
Worarat Kruasuwan ◽  
Aekkachai Puseenam ◽  
Sutipa Tanapongpipat ◽  
Niran Roongsawang
Keyword(s):  
Protein Production ◽  
Secretory Pathway ◽  
Genome Engineering ◽  
Heterologous Protein ◽  
Methylotrophic Yeast ◽  
Strain Engineering ◽  
Heterologous Protein Production ◽  
Heterologous Protein Secretion ◽  
Multiple Genes ◽  
Simultaneous Activation

CRISPR multiplex gRNA systems have been employed in genome engineering in various industrially relevant yeast species. The thermotolerant methylotrophic yeast Ogataea thermomethanolica TBRC 656 is an alternative host for heterologous protein production. However, the limited secretory capability of this yeast is a bottleneck for protein production. Here, we refined CRISPR-based genome engineering tools for simultaneous mutagenesis and activation of multiple protein secretory pathway genes to improve heterologous protein secretion. We demonstrated that multiplexed CRISPR-Cas9 mutation of up to four genes (SOD1, VPS1, YPT7 and YPT35) in one single cell is practicable. We also developed a multiplexed CRISPR-dCas9 system which allows simultaneous activation of multiple genes in this yeast. 27 multiplexed gRNA combinations were tested for activation of three genes (SOD1, VPS1 and YPT7), three of which were demonstrated to increase the secretion of fungal xylanase and phytase up to 29% and 41%, respectively. Altogether, our study provided a toolkit for mutagenesis and activation of multiple genes in O. thermomethanolica, which could be useful for future strain engineering to improve heterologous protein production in this yeast.

scholarly journals Optimizing heterologous protein production in the periplasm of E. coli by regulating gene expression levels

2013 ◽  
Vol 12 (1) ◽  
pp. 24 ◽  
Author(s):  
Susan Schlegel ◽  
Edurne Rujas ◽  
Anders Jimmy Ytterberg ◽  
Roman A Zubarev ◽  
Joen Luirink ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production ◽  
Expression Levels ◽  
E Coli ◽  
Gene Expression Levels

Identification of proteins responsive to heterologous protein production in thermotolerant methylotrophic yeast Ogataea thermomethanolica TBRC656

Yeast ◽  
2021 ◽  
Author(s):  
Chitwadee Phithakrotchanakoon ◽  
Aekkachai Puseenam ◽  
Worarat Kruasuwan ◽  
Somsak Likhitrattanapisal ◽  
Narumon Phaonakrop ◽  
...  
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Methylotrophic Yeast ◽  
Heterologous Protein Production

New Promoters to Improve Heterologous Protein Production in Aspergillus vadensis

2014 ◽  
Vol 3 (3) ◽  
pp. 244-251 ◽  
Author(s):  
Helena Culleton ◽  
Ourdia Bouzid ◽  
Vincent McKie ◽  
Ronald Vries
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production
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