The effect of pre- and pro-sequences and multicopy integration on heterologous expression of the Fusarium solani pisi cutinase gene in Aspergillus awamori

Applied Microbiology and Biotechnology
I A van GemerenC T Verrips

Abstract

A synthetic derivative of the cutinase cDNA of Fusarium solani pisi was expressed in Aspergillus awamori using the A. awamori endoxylanase II (exlA) promoter and terminator. The influence of the origin of the pre-sequence and the presence of a pro-sequence on the efficiency of extracellular cutinase production was analysed in single-copy transformants containing an expression cassette integrated at the pyrG locus. Transformants containing a construct encoding a direct, inframe fusion of the xylanase pre-peptide to the mature cutinase showed a 2-fold higher cutinase production level compared to strains containing constructs with an additional cutinase pro-peptide. The effect of multicopy integration of the expression cassette on cutinase production was analysed in strains with different numbers of a cutinase construct containing its own pre-pro-sequence. The multicopy strains showed a 6-to 12-fold increased production of extracellular cutinase relative to the single-copy strains. No linear dose response relation to the number of expression cassettes present in the strains was observed. The amount of active enzyme produced by the strains correlated with the amount of cutinase-specific mRNA, suggesting that cutinase overproduction...Continue Reading

Citations

Jan 17, 2007·Applied Microbiology and Biotechnology·Sriappareddy TamalampudiHideki Fukuda
Jun 23, 2000·Enzyme and Microbial Technology·T VerripsabC Sagt
Jun 20, 2003·Journal of Biotechnology·B Christien LokmanCees A M J J van den Hondel
Dec 2, 2000·Biochimie·M R Egmond, J de Vlieg
May 26, 2001·Applied and Environmental Microbiology·R J GoukaC T Verrips
Dec 9, 2008·Bioscience, Biotechnology, and Biochemistry·Yali ChengRichard R Bélanger
Sep 21, 2004·Applied Microbiology and Biotechnology·Vivi JoostenB Christien Lokman
Jan 1, 1997·Critical Reviews in Biotechnology·D B Archer, J F Peberdy
Jul 8, 2009·Protein Expression and Purification·Min-A KwonJae Kwang Song
Oct 9, 2008·Biotechnology Advances·David Lubertozzi, Jay D Keasling
Aug 10, 2001·Fungal Genetics and Biology : FG & B·A ConesaC A van den Hondel
Sep 24, 2013·Biotechnology Advances·Sheng ChenJing Wu
Mar 18, 2006·Advances in Applied Microbiology·O P WardA Singh
Nov 11, 1999·Biotechnology and Bioengineering·C M CarvalhoJ M Cabral
Aug 28, 2014·Critical Reviews in Microbiology·Monika Sharma, Rohit Sharma
Aug 4, 1998·Applied and Environmental Microbiology·I A van GemerenC T Verrips
Sep 3, 1998·Applied and Environmental Microbiology·S P GollerC P Kubicek

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