Synergistic co-utilization of biomass-derived sugars enhances aromatic amino acid production by engineered Escherichia coli.

Autor: Liu A; Biological Design Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Machas M; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Mhatre A; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Hajinajaf N; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Sarnaik A; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Nichols N; US Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, Illinois, USA., Frazer S; US Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, Illinois, USA., Wang X; Biological Design Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA.; School of Life Sciences, Arizona State University, Tempe, Arizona, USA., Varman AM; Biological Design Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA.; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA., Nielsen DR; Biological Design Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA.; Chemical Engineering Program, School for Engineering of Matter, Transport, and Energy, Arizona State University, Tempe, Arizona, USA.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2024 Feb; Vol. 121 (2), pp. 784-794. Date of Electronic Publication: 2023 Nov 05.
DOI: 10.1002/bit.28585
Abstrakt: Efficient co-utilization of mixed sugar feedstocks remains a biomanufacturing challenge, thus motivating ongoing efforts to engineer microbes for improved conversion of glucose-xylose mixtures. This study focuses on enhancing phenylalanine production by engineering Escherichia coli to efficiently co-utilize glucose and xylose. Flux balance analysis identified E4P flux as a bottleneck which could be alleviated by increasing the xylose-to-glucose flux ratio. A mutant copy of the xylose-specific activator (XylR) was then introduced into the phenylalanine-overproducing E. coli NST74, which relieved carbon catabolite repression and enabled efficient glucose-xylose co-utilization. Carbon contribution analysis through 13 C-fingerprinting showed a higher preference for xylose in the engineered strain (NST74X), suggesting superior catabolism of xylose relative to glucose. As a result, NST74X produced 1.76 g/L phenylalanine from a model glucose-xylose mixture; a threefold increase over NST74. Then, using biomass-derived sugars, NST74X produced 1.2 g/L phenylalanine, representing a 1.9-fold increase over NST74. Notably, and consistent with the carbon contribution analysis, the xylR* mutation resulted in a fourfold greater maximum rate of xylose consumption without significantly impeding the maximum rate of total sugar consumption (0.87 vs. 0.70 g/L-h). This study presents a novel strategy for enhancing phenylalanine production through the co-utilization of glucose and xylose in aerobic E. coli cultures, and highlights the potential synergistic benefits associated with using substrate mixtures over single substrates when targeting specific products.
(© 2023 Wiley Periodicals LLC.)
Databáze: MEDLINE
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