A gene replacement strategy for engineering nisin.

Microbiology
Helen M DoddMichael J Gasson

Abstract

A lactococcal expression system was developed which allows the exclusive production of novel nisins encoded by mutated pre-nisin (nisA) genes. This system is based on a combination of a specifically constructed host strain and vectors which facilitate the genetic manipulation of the nisA gene. The wild-type chromosomal gene is effectively replaced with a variant nisA gene, by the technique of gene replacement. The recovery of full nisin immunity was employed as a means of directly selecting strains that had acquired an intact nisA gene by the gene replacement process. With this approach the other genes required for pre-nisin maturation are not affected and any alterations to DNA sequences are restricted to only those specific mutations introduced in the nisA gene. The effectiveness of the system was demonstrated by the expression of a number of variant nisA genes leading to the successful production and characterization of nisins containing the substitutions Dha5A, Dha33A, Dha5, 33A, H27K, 130W and K12L. The enhanced yields of these engineered nisin molecules, when compared to their production in a plasmid-complementation system, underlines the improvement offered by this gene replacement strategy.

References

Nov 1, 1992·Applied and Environmental Microbiology·H M DoddM J Gasson
Jan 1, 1992·Applied and Environmental Microbiology·C KleinK D Entian
Aug 1, 1991·Molecular & General Genetics : MGG·N HornM Gasson
Nov 1, 1991·European Journal of Biochemistry·J W MuldersW M de Vos
Mar 1, 1990·Journal of General Microbiology·H M DoddM J Gasson
Feb 1, 1989·Applied and Environmental Microbiology·K J LeenhoutsG Venema
Jul 1, 1989·Applied and Environmental Microbiology·M C ChopinN Galleron
Apr 1, 1980·Journal of Molecular Biology·M J Casadaban, S N Cohen
Aug 1, 1995·Applied and Environmental Microbiology·H S RollemaR J Siezen
Nov 10, 1995·The Journal of Biological Chemistry·O P KuipersW M de Vos
Mar 1, 1995·Applied and Environmental Microbiology·K Siegers, K D Entian
Aug 1, 1994·Applied and Environmental Microbiology·C Klein, K D Entian
Mar 1, 1994·Applied and Environmental Microbiology·G EngelkeK D Entian
Jan 1, 1993·Applied and Environmental Microbiology·C KleinK D Entian
Sep 1, 1990·Applied and Environmental Microbiology·K J LeenhoutsG Venema
Jun 1, 1975·Applied Microbiology·B E Terzaghi, W E Sandine

❮ Previous
Next ❯

Citations

May 3, 2005·International Journal of Food Microbiology·R Bauer, L M T Dicks
Jan 14, 2000·International Journal of Food Microbiology·N A OlasupoW H Holzapfel
Apr 24, 1999·European Journal of Biochemistry·A Karakas SenM J Gasson
Feb 3, 1999·Annals of the New York Academy of Sciences·H KimuraA Ishizaki
Jul 4, 2003·Applied and Environmental Microbiology·Christiane SzekatGabriele Bierbaum
Oct 1, 1996·Antonie van Leeuwenhoek·M E Stiles
Nov 26, 2010·Molecular Microbiology·Des FieldR Paul Ross
Dec 29, 2005·Biotechnology Advances·Xu Xia ZhouYuan Jiang Pan
Jan 1, 1997·Journal of Bacteriology·G N MollA J Driessen
Aug 1, 1996·Applied and Environmental Microbiology·W C ChanG C Roberts

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antifungals

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antifungals (ASM)

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.