- Autor
- Mellitzer, Andrea
- Weis, Roland
- Glieder, Anton
- Flicker, Karlheinz
- TitelExpression of lignocellulolytic enzymes in Pichia pastoris
- Datei
- DOI10.1186/1475-2859-11-61
- Persistent Identifier
- Erschienen inMicrobial Cell Factories
- Band11
- Erscheinungsjahr2012
- Heft1
- LicenceCC-BY
- ISSN1475-2859
- Download Statistik1666
- Peer ReviewNein
- AbstractBACKGROUND: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.
[598.79 KB]