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

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.

The Methylotrophic Yeast Pichia Pastoris as a Host for Heterologous Protein Production

1987 ◽  
pp. 289-296 ◽  
Author(s):  
G. Thill ◽  
G. Davis ◽  
C. Stillmann ◽  
J. F. Tschopp ◽  
W. S. Graig ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Protein Production ◽  
Heterologous Protein ◽  
Methylotrophic Yeast ◽  
Heterologous Protein Production ◽  
Yeast Pichia Pastoris ◽  
Methylotrophic Yeast Pichia Pastoris

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

Absence of See1p, a widely conservedSaccharomyces cerevisiaeprotein, confers both deficient heterologous protein production and endocytosis

Yeast ◽  
2008 ◽  
Vol 25 (12) ◽  
pp. 871-877 ◽  
Author(s):  
Cristina Martín-Granados ◽  
Sean-Patrick Riechers ◽  
Ulf Stahl ◽  
Christine Lang
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production

scholarly journals State and specific growth estimation in heterologous protein production by Pichia pastoris

AIChE Journal ◽  
2011 ◽  
Vol 58 (10) ◽  
pp. 2966-2979 ◽  
Author(s):  
José M. Barrigón ◽  
Ramon Ramon ◽  
Isabel Rocha ◽  
Francisco Valero ◽  
Eugénio C. Ferreira ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Protein Production ◽  
Heterologous Protein ◽  
Specific Growth ◽  
Heterologous Protein Production ◽  
Growth Estimation

Heterologous protein production using thePichia pastoris expression system

Yeast ◽  
2005 ◽  
Vol 22 (4) ◽  
pp. 249-270 ◽  
Author(s):  
Sue Macauley-Patrick ◽  
Mariana L. Fazenda ◽  
Brian McNeil ◽  
Linda M. Harvey
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Expression System ◽  
Heterologous Protein Production ◽  
Pastoris Expression System

Biotechnological Exploitation of Heterologous Protein Production in Fungi

Fungi ◽  
2017 ◽  
pp. 227-250
Author(s):  
Brendan Curran ◽  
Virginia Bugeja
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Heterologous Protein Production
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