Expression of lignocellulolytic enzymes in Pichia pastoris.

Microbial Cell Factories
A MellitzerKarlheinz Flicker

Abstract

Sustainable utilization of plant biomass as renewable source for fuels and chemical building blocks requires a complex mixture of diverse enzymes, including hydrolases which comprise the largest class of lignocellulolytic enzymes. These enzymes need to be available in large amounts at a low price to allow sustainable and economic biotechnological processes.Over the past years Pichia pastoris has become an attractive host for the cost-efficient production and engineering of heterologous (eukaryotic) proteins due to several advantages. In this paper codon optimized genes and synthetic alcohol oxidase 1 promoter variants were used to generate Pichia pastoris strains which individually expressed cellobiohydrolase 1, cellobiohydrolase 2 and beta-mannanase from Trichoderma reesei and xylanase A from Thermomyces lanuginosus. For three of these enzymes we could develop strains capable of secreting gram quantities of enzyme per liter in fed-batch cultivations. Additionally, we compared our achieved yields of secreted enzymes and the corresponding activities to literature data. In our experiments we could clearly show the importance of gene optimization and strain characterization for successfully improving secretion levels. We also pres...Continue Reading

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Citations

Nov 6, 2013·Microbial Cell Factories·Ewelina Celińska, Włodzimierz Grajek
Apr 20, 2014·Microbial Cell Factories·Anikó VárnaiVincent G H Eijsink
Jul 16, 2014·Critical Reviews in Biotechnology·Montarop YamabhaiDietmar Haltrich
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Apr 19, 2015·Microbial Cell Factories·Pengfei LiTaicheng Zhu
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May 16, 2020·Frontiers in Bioengineering and Biotechnology·Moira GiovannoniBenedetta Mattei
Jul 7, 2020·Frontiers in Bioengineering and Biotechnology·Lingkai ZhangJian Li
Dec 14, 2017·Chembiochem : a European Journal of Chemical Biology·Veeresh Juturu, Jin Chuan Wu
Oct 21, 2020·Scientific Reports·Hongguang FuZhong Liu
Jul 2, 2021·Microbial Cell Factories·Aravind MadhavanParameswaran Binod

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Methods Mentioned

BETA
protein folding
glycosylation
PCR
electrophoresis

Software Mentioned

Gene Designer

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