A lactococcal expression system for engineered nisins

• Published in: Applied and Environmental Microbiology 1992-11, Vol.58 (11), p.3683-3693
• Main Authors: Dodd, H.M. (AFRC Institute of Food Research, Norwich, UK), Horn, N, Hao, Z, Gasson, M.J
• Format: Article
• Language: English
• Subjects: ADN RECOMBINADO; ADN RECOMBINE; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Biology; Biotechnology; CHROMOSOME; Cloning, Molecular; CROMOSOMAS; Drug Resistance, Microbial; EXPRESION GENICA; EXPRESSION DES GENES; Fundamental and applied biological sciences. Psychology; Genes, Bacterial - genetics; Genetic engineering; Genetic technics; Genetic Vectors - genetics; Genetics; LACTOCOCCUS LACTIS; Lactococcus lactis - genetics; Methods. Procedures. Technologies; Molecular Sequence Data; Molecules; MUTACION; Mutagenesis, Insertional; MUTATION; Nisin - biosynthesis; Nisin - genetics; NISINA; NYSINE; Plasmids - genetics; Recombinant Proteins - metabolism; SECUENCIA NUCLEICA; SEQUENCE NUCLEIQUE; Transcription, Genetic; Vectors (cloning, transfer, expression). Insertion sequences and transposons
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.58.11.3683-3693.1992

The nisin-producing Lactococcus lactis strain F15876 has been modified and developed for use as an expression system for engineered nisin variants. Insertional inactivation of the resident nisA gene had a polar effect on downstream genes, including those involved in nisin immunity. However, subsequent chromosomal rearrangements in this region involving a newly discovered insertion element (IS905) generated a strain that was deficient in the nisA gene product but expressed those nisin determinants necessary for prenisin maturation, secretion, and immunity. Complementation of the lesion in the nisA gene by plasmid-encoded nisA genes containing site-specific mutations resulted in the exclusive production of altered nisins containing specific amino acid substitutions