Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 110, Issue 10, Pages 2616-2623Publisher
WILEY-BLACKWELL
DOI: 10.1002/bit.24938
Keywords
lignocellulosic biomass; HMF; furfural; acetic acid; evolutionary engineering; complex phenotypes; genomics
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Funding
- US NSF [CBET-1032487, MCB-1054276]
- Directorate For Engineering
- Div Of Chem, Bioeng, Env, & Transp Sys [1032487] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [1054276] Funding Source: National Science Foundation
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Lignocellulosic biomass has become an important feedstock to mitigate current ethical and economical concerns related to the bio-based production of fuels and chemicals. During the pre-treatment and hydrolysis of the lignocellulosic biomass, a complex mixture of sugars and inhibitors are formed. The inhibitors interfere with microbial growth and product yields. This study uses an adaptive laboratory evolution method called visualizing evolution in real-time (VERT) to uncover the molecular mechanisms associated with tolerance to hydrolysates of lignocellulosic biomass in Saccharomyces cerevisiae. VERT enables a more rational scheme for isolating adaptive mutants for characterization and molecular analyses. Subsequent growth kinetic analyses of the mutants in individual and combinations of common inhibitors present in hydrolysates (acetic acid, furfural, and hydroxymethylfurfural) showed differential levels of resistance to different inhibitors, with enhanced growth rates up to 57%, 12%, 22%, and 24% in hydrolysates, acetic acid, HMF and furfural, respectively. Interestingly, some of the adaptive mutants exhibited reduced fitness in the presence of individual inhibitors, but showed enhanced fitness in the presence of combinations of inhibitors compared to the parental strains. Transcriptomic analysis revealed different mechanisms for resistance to hydrolysates and a potential cross adaptation between oxidative stress and hydrolysates tolerance in several of the mutants. Biotechnol. Bioeng. 2013;110: 2616-2623. (c) 2013 Wiley Periodicals, Inc.
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