Comparative genomics revealed key molecular targets to rapidly convert a reference rifamycin-producing bacterial strain into an overproducer by genetic engineering.
Metab Eng, 2014/11;26:1-16.
Peano C[1], Damiano F[2], Forcato M[3], Pietrelli A[1], Palumbo C[2], Corti G[1], Siculella L[2], Fuligni F[1], Tagliazucchi GM[3], De Benedetto GE[4], Bicciato S[3], De Bellis G[1], Alifano P[5]
Affiliations
PMID: 25149266DOI: 10.1016/j.ymben.2014.08.001
Impact factor: 8.829
Abstract
Rifamycins are mainstay agents in treatment of many widespread diseases, but how an improved rifamycin producer can be created is still incompletely understood. Here, we describe a comparative genomic approach to investigate the mutational patterns introduced by the classical mutate-and-screen method in the genome of an improved rifamycin producer. Comparing the genome of the rifamycin B overproducer Amycolatopsis mediterranei HP-130 with those of the reference strains A. mediterranei S699 and U32, we identified 250 variations, affecting 227 coding sequences (CDS), 109 of which were HP-130-specific since they were absent in both S699 and U32. Mutational and transcriptional patterns indicated a series of genomic manipulations that not only proved the causative effect of mutB2 (coding for methylmalonyl-CoA mutase large subunit) and argS2 (coding for arginyl tRNA synthetase) mutations on the overproduction of rifamycin, but also constituted a rational strategy to genetically engineer a reference strain into an overproducer.
Keywords: Amycolatopsis mediterranei; Arginyl tRNA synthetase; Methylmalonyl-CoA mutase; Rifamycin; Strain improvement
MeSH terms
Actinobacteria; Arginine-tRNA Ligase; Chromosome Mapping; Comparative Genomic Hybridization; Gene Targeting; Genetic Enhancement; Genome, Bacterial; Metabolic Engineering; Methylmalonyl-CoA Mutase; Rifamycins; Species Specificity; Up-Regulation
More resources
EndNote: Download