PMID: 2111228Mar 1, 1990Paper

Cloning and expression of a hybrid Streptomyces clavuligerus cefE gene in Penicillium chrysogenum

Current Genetics
C A CantwellS W Queener

Abstract

A hybrid cefE gene was constructed by juxtaposing promoter sequences from the Penicillium chrysogenum pcbC gene to the open reading frame of the Streptomyces clavuligerus cefE gene. In S. clavuligerus the cefE gene codes for the enzyme penicillin N expandase [also known as deacetoxycephalosporin C synthetase (DAOCS)]. To insert the hybrid cefE gene into P. chrysogenum the vector pPS65 was constructed; pPS65 contains the hybrid cefE gene and the Aspergillus nidulans amdS gene. The amdS gene encodes acetamidase and provides for dominant selection in P. chrysogenum. Protoplasts of P. chrysogenum were transformed with pPS65 and selected for the ability to grow on acetamide medium. Extracts of cells cultivated in penicillin production medium were analyzed for penicillin N expandase activity. Penicillin N expandase activity was detected in approximately one-third of the transformants tested. Transformants WG9-69C-01 and WG9-61L-03 had the highest specific activities of penicillin N expandase: 4.3% and 10.3%, respectively, relative to the amount of penicillin N expandase in S. clavuligerus. Untransformed P. chrysogenum exhibited no penicillin N expandase activity. Analysis of the penicillin V titer revealed that WG9-61L-03 produced ti...Continue Reading

References

Jan 1, 1987·Current Genetics·P L SkatrudD L Fisher
Mar 23, 1968·Nature·M Meselson, R Yuan

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Citations

Jan 1, 1990·Annals of the New York Academy of Sciences·W K Yeh, S W Queener
May 2, 1994·Annals of the New York Academy of Sciences·S W QueenerP Rosteck
Apr 8, 2003·Applied Microbiology and Biotechnology·R P Elander
Aug 2, 2011·Applied Microbiology and Biotechnology·Marco Alexander van den Berg
Feb 10, 2012·Journal of Biomedicine & Biotechnology·Carlos BarreiroCarlos García-Estrada
Sep 3, 2013·Fungal Genetics and Biology : FG & B·Paulo de BoerRemko Offringa
Nov 9, 2016·Journal of Industrial Microbiology & Biotechnology·Keqiang FanKeqian Yang
Jan 1, 1994·Critical Reviews in Biotechnology·W Piepersberg
Jul 5, 1997·Biotechnology and Bioengineering·B DiezJ L Barredo
Sep 30, 2008·Nature Biotechnology·Marco A van den BergRoel A L Bovenberg
May 12, 2019·Transfusion·Toshihiro HorikawaMitsunobu Tanaka
Jan 1, 2010·Biotechnology & Genetic Engineering Reviews·Marco Van Den BergIda Vander Keli
May 5, 2020·Natural Product Reports·Stephanie GalanieJames Lalonde
Nov 9, 2012·Natural Product Reports·Refaat B HamedChristopher J Schofield
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·A A Brakhage

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