Journal article
Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose by Clostridium thermocellum at elevated temperatures
Biotechnology for biofuels, Vol.12(1), pp.245-245
10/15/2019
DOI: 10.1186/s13068-019-1583-8
PMCID: PMC6792240
PMID: 31636704
Abstract
Background: Esters are versatile chemicals and potential drop-in biofuels. To develop a sustainable production platform, microbial ester biosynthesis using alcohol acetyltransferases (AATs) has been studied for decades. Volatility of esters endows high-temperature fermentation with advantageous downstream product separation. However, due to the limited thermostability of AATs known, the ester biosynthesis has largely relied on use of mesophilic microbes. Therefore, developing thermostable AATs is important for ester production directly from lignocellulosic biomass by the thermophilic consolidated bioprocessing (CBP) microbes, e.g., Clostridium thermocellum.
Results: In this study, we engineered a thermostable chloramphenicol acetyltransferase from Staphylococcus aureus (CAT(Sa)) for enhanced isobutyl acetate production at elevated temperatures. We first analyzed the broad alcohol substrate range of CAT(Sa). Then, we targeted a highly conserved region in the binding pocket of CAT(Sa) for mutagenesis. The mutagenesis revealed that F97W significantly increased conversion of isobutanol to isobutyl acetate. Using CAT(Sa) F97W, we demonstrated direct conversion of cellulose into isobutyl acetate by an engineered C. thermocellum at elevated temperatures.
Conclusions: This study highlights that CAT is a potential thermostable AAT that can be harnessed to develop the thermophilic CBP microbial platform for biosynthesis of designer bioesters directly from lignocellulosic biomass.
Details
- Title: Subtitle
- Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose by Clostridium thermocellum at elevated temperatures
- Creators
- Hyeongmin Seo - University of Tennessee at KnoxvilleJong-Won Lee - University of Tennessee at KnoxvilleSergio Garcia - University of Tennessee at KnoxvilleCong T. Trinh - University of Tennessee at Knoxville
- Resource Type
- Journal article
- Publication Details
- Biotechnology for biofuels, Vol.12(1), pp.245-245
- Publisher
- Springer Nature
- DOI
- 10.1186/s13068-019-1583-8
- PMID
- 31636704
- PMCID
- PMC6792240
- ISSN
- 1754-6834
- eISSN
- 1754-6834
- Number of pages
- 13
- Grant note
- U.S. Department of Energy (DOE) Bioenergy Research Centers - Office of Biological and Environmental Research in the DOE Office of Science; United States Department of Energy (DOE) 1553250 / NSF CAREER award (NSF); National Science Foundation (NSF) DE-AC02-05CH11231 / Office of Science of the U.S. Department of Energy; United States Department of Energy (DOE) Center for Bioenergy Innovation (CBI)
- Language
- English
- Date published
- 10/15/2019
- Academic Unit
- Chemical and Biochemical Engineering
- Record Identifier
- 9984696579602771
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