Production of chemicals via fermentation has been evolving over the past 30 years in search of economically viable systems. Thus far, there have been few industrially relevant chemicals that have seen commercialization, examples being lactic acid and ethanol. Currently, many of these fermentation processes still compete with food sources. In order to reduce this competition fermentation of alternative feedstocks, such as lignocellulosic biomass must to be utilized. Hemicellulosic sugars can be employed effectively for the production of chemicals by incorporating nonphosphorylative metabolism. This review covers nonphosphorylative metabolism, the pathways and enzymes involved, as well as the products that have been produced using nonphosphorylative metabolism.
Vancomycin-resistant Enterococci infections are a significant clinical problem. One proposed solution is to use probiotics, such as lactic acid bacteria, to produce antimicrobial peptides at the site of infection. Enterocin A, a class 2a bacteriocin, exhibits inhibitory activity against E. faecium and E. faecalis, which account for 86% of vancomycin-resistant Enterococci infections. In this study, we aimed to engineer enterocin A mutants with enhanced potency within a lactic acid bacterial production system. Peptide mutants resulting from saturation mutagenesis at sites A24 and T27 were efficiently screened in a 96-well plate assay for inhibition of pathogen growth. Several mutants exhibit increased potency relative to wild-type enterocin A in both liquid- and solid-medium growth assays. In particular, A24P and T27G exhibit enhanced inhibition of multiple strains of E. faecium and E. faecalis, including clinically isolated vancomycin-resistant strains. A24P and T27G enhance killing of E. faecium 8 by 13±3- and 18±4-fold, respectively. The engineered enterocin A/lactic acid bacteria systems offer significant potential to combat antibiotic-resistant infections.
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