Metabolic pathway optimization for biosynthesis of 1,2,4-butanetriol from xylose by engineered Escherichia coli |
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| Institution: | 1. Green Chemistry and Materials Group, Korea Institute of Industrial Technology, Cheonan si, Chungcheongnam-do, 31056, Republic of Korea;2. Green Process and System Engineering Major, Korea University of Science and Technology (UST), Daejeon, 34141, Republic of Korea;3. STR Biotech Co., Ltd., Bioplaza 4-3, 56, Soyanggang-ro, Chuncheon-si, Gangwon-do, 200-957, Republic of Korea;4. Department of Molecular Bio-science, Kangwon National University, Chuncheon-si, Gangwon-do, 200-701, Republic of Korea;5. Department of Biological Engineering, Inha University, Incheon, 402-751, Republic of Korea;1. Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN 55455, USA;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;3. College of Life Science, Shanxi Normal University, Linfen 041004, China;1. Department of Chemical Engineering and Materials Science, Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea;2. Department of Chemical Engineering, Kwangwoon University, Seoul 01897, Republic of Korea;1. College of Engineering, University of Georgia, Athens, GA 30602, USA;2. State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China;3. BioChemical Engineering Program, College of Engineering, University of Georgia, Athens, GA 30602, USA |
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| Abstract: | 1,2,4-Butanetriol (BT) and related derivatives have been widely used in many fields, especially in the military and in medicine. In this paper, we systematically optimized the BT biosynthetic pathway. We first investigated the activities of various NADH dependent aldehyde reductases (ALRs), which catalyze the fourth reaction in the four-step pathway for BT production from xylose in E. coli, and found that a combination of multiple endogenous enzymes catalyzed aldehyde reduction in the BT production bioprocess and that YqhD in E. coli was a main ALR for BT production. In addition, ADH2 from Saccharomyces cerevisiae can effectively catalyze 3,4-dihydroxybutanal to BT. Also, YjhG was identified as the major xylonate dehydratase and was co-overexpressed with YqhD, resulting in an improvement of BT production by 30%. Moreover, we identified and eliminated the competing branch pathway by inactivating 2-keto acid reductases (yiaE). Finally, the combination of these approaches led to BT production of 5.1 g/L. In summary, our study provides insights into the biosynthetic pathway for BT production, demonstrates an effective strategy to enhance BT production, and paves the way toward in-depth research on BT biosynthesis. |
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| Keywords: | 1 2 4-Butanetriol metabolic pathway aldehyde reductase xylonate dehydratase branch pathway |
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