Abstract
BACKGROUND: 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.
RESULTS: 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.
CONCLUSION: In our experiments we could clearly show the importance of gene optimization and strain characterization for successfully improving secretion levels. We also present a basic guideline how to correctly interpret the interplay of promoter strength and gene dosage for a successful improvement of the secretory production of lignocellulolytic enzymes in Pichia pastoris.
Original language | English |
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Article number | 61 |
Number of pages | 11 |
Journal | Microbial Cell Factories |
Volume | 11 |
DOIs | |
Publication status | Published - 14 May 2012 |
Keywords
- Alcohol Oxidoreductases
- Cellulose 1,4-beta-Cellobiosidase
- Endo-1,4-beta Xylanases
- Fermentation
- Gene Dosage
- Gene Expression Regulation, Fungal
- Pichia
- Promoter Regions, Genetic
- Recombinant Proteins
- Trichoderma
- beta-Mannosidase
- Journal Article
- Research Support, Non-U.S. Gov't