Strain improvement for cephalosporin production by Acremonium chrysogenum using geneticin as a suitable transformation marker.

Abstract:

:An Acremonium chrysogenum strain improvement program based on the transformation with cephalosporin biosynthetic genes was carried out to enhance cephalosporin C production. Best results were obtained with cefEF and cefG genes, selecting transformants with increased cephalosporin C production and lower accumulation of biosynthetic intermediates. Phleomycin resistant transformants, designated B1 and C1, showed a single copy random integration event, higher levels of cefEF transcript and, according to immunoblotting analyses, higher amounts of deacetylcephalosporin C acetyltransferase (DAC-AT) protein than their parental strains. Moreover, DAC-AT activity was higher in the transformants. Plasmids carrying geneticin resistance markers based on the nptII gene from Tn5 and the aphI gene from Tn903 were constructed to transform again B1 and C1, showing that the cassette Pgdh-nptII-trpC was able to confer geneticin resistance to A. chrysogenum and demonstrating that geneticin is a helpful selection marker.

journal_name

FEMS Microbiol Lett

authors

Rodríguez-Sáiz M,Lembo M,Bertetti L,Muraca R,Velasco J,Malcangi A,de la Fuente JL,Barredo JL

doi

10.1016/j.femsle.2004.04.010

keywords:

subject

Has Abstract

pub_date

2004-06-01 00:00:00

pages

43-9

issue

1

eissn

0378-1097

issn

1574-6968

pii

S0378109704002691

journal_volume

235

pub_type

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