Gelatin expression from an engineered Saccharomyces cerevisiae CUP1 promoter in Pichia pastoris
- 12 February 2021
- Vol. 38 (6), 382-387
- https://doi.org/10.1002/yea.3554
Abstract
The methylotrophic yeast Pichia pastoris (reclassified as Komagataella phaffii) is a versatile protein expression system, yet many commonly used promoters have attributes undesirable for fermentation or its optimization. Hence, the copper‐inducible CUP1 gene promoter from the related yeast Saccharomyces cerevisiae was used to express human gelatin. Multimerization of a potential copper response element in the CUP1 promoter, a S. cerevisiae Ace1p binding site, significantly increased gelatin expression. Expression was induced by copper in a dose‐dependent fashion and was not dependent on cell density. Gelatin was additionally induced in standard copper‐containing fermentation basal salts media. Removal of a S. cerevisiae heat shock factor (Hsf1p) binding site reduced copper‐dependent gelatin induction suggesting that a similar protein may regulate this promoter in P. pastoris. This engineered copper inducible promoter expands the yeast recombinant protein production tool kit.Keywords
Funding Information
- FibroGen
This publication has 23 references indexed in Scilit:
- Genome sequence of the recombinant protein production host Pichia pastorisNature Biotechnology, 2009
- Expression and characterization of a low molecular weight recombinant human gelatin: development of a substitute for animal-derived gelatin with superior featuresProtein Expression and Purification, 2005
- Heterologous protein expression in the methylotrophic yeastPichia pastorisFEMS Microbiology Reviews, 2000
- Applications of yeast in biotechnology: protein production and genetic analysisCurrent Opinion in Biotechnology, 1999
- Isolation of the Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase gene and regulation and use of its promoterGene, 1997
- Construction of a CUP1 promoter-based vector to modulate gene expression in Saccharomyces cerevisiaeGene, 1996
- Distinct Regions of Cu(I)·ACE1 Contact Two Spatially Resolved DNA Major Groove SitesOnline Journal of Public Health Informatics, 1995
- Heat shock factor and the heat shock responseCell, 1991
- Versatile cassettes designed for the copper inducible expression of proteins in yeastPlasmid, 1989
- Cloning and expression of a yeast copper metallothionein geneGene, 1984