Abstract
1,2,4-Butanetriol (BT) is an important non-natural chemical with a variety of industrial applications. Identifying the bottlenecks for BT biosynthesis is expected to contribute to improving the efficiency of this process. In this work, we first constructed a prototype strain for BT production by assembling a four-step synthetic pathway and disrupting the competing pathways for xylose in Escherichia coli BW25113. Using this prototype strain, we conducted systematic fine-tuning of the pathway enzyme expression level to identify the potential bottlenecks and optimize the BT biosynthesis. Production conditions were also optimized by exploring the effects of temperature, pH and cell density on BT titer. BT production was increased by 4.3-fold from the prototype strain, achieved a final titer of 1.58 g/L with a yield of 7.9 % after 72-h biotransformation.
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References
Abdel-Ghany SE, Day I, Heuberger AL, Broeckling CD, Reddy ASN (2013) Metabolic engineering of Arabidopsis for butanetriol production using bacterial genes. Metab Eng 20:109–120. doi:10.1016/j.ymben.2013.10.003
Amann E, Brosius J, Ptashne M (1983) Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene 25:167–178. doi:10.1016/0378-1119(83)90222-6
Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2(2006):0008. doi:10.1038/msb4100050
Cao Y, Xian M, Zou H, Zhang H (2013) Metabolic engineering of Escherichia coli for the production of xylonate. PLoS One 8:e67305. doi:10.1371/journal.pone.0067305
Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645. doi:10.1073/pnas.120163297
Davis JH, Rubin AJ, Sauer RT (2011) Design, construction and characterization of a set of insulated bacterial promoters. Nucl Acids Res 39:1131–1141. doi:10.1093/Nar/Gkq810
de Boer HA, Comstock LJ, Vasser M (1983) The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci USA 80:21–25
Frost JW, Niu W (2008) Microbial synthesis of d-1,2,4-butanetriol. WO2008091288 A2, Mar 19, 2009
Gouranlou F, Kohsary I (2010) Synthesis and characterization of 1,2,4-butanetrioltrinitrate. Asian J Chem 22:4221–4228
Jiang Y, Liu W, Cheng T, Cao Y, Zhang R, Xian M (2015) Characterization of d-xylonate dehydratase YjhG from Escherichia coli. Bioengineered 6:227–232. doi:10.1080/21655979.2015.1040208
Levin-Karp A, Barenholz U, Bareia T, Dayagi M, Zelcbuch L, Antonovsky N, Noor E, Milo R (2013) Quantifying translational coupling in E. coli synthetic operons using RBS modulation and fluorescent reporters. ACS Synth Biol 2:327–336. doi:10.1021/Sb400002n
Li XH, Cai Z, Li Y, Zhang YP (2014) Design and construction of a non-natural malate to 1,2,4-butanetriol pathway creates possibility to produce 1,2,4-butanetriol from glucose. Sci Rep UK. doi:10.1038/Srep05541
Lou CB, Stanton B, Chen YJ, Munsky B, Voigt CA (2012) Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nat Biotechnol 30:1137-+. doi:10.1038/Nbt.2401
Niu W, Molefe MN, Frost JW (2003) Microbial synthesis of the energetic material precursor 1,2,4-butanetriol. J Am Chem Soc 125:12998–12999. doi:10.1021/Ja036391
Ren T, Liu DX (1999) Synthesis of cationic lipids from 1,2,4-butanetriol. Tetrahedron Lett 40:209–212. doi:10.1016/S0040-4039(98)02381-8
Sun L, Yang F, Zhu T, Li X, Sun H, Li Y, Xu Z, Zhang Y (2015) Optimization of 1,2,4-butanetriol synthetic pathway in Escherichia coli. Chin J Biotech (in press)
Tandon VK, Van Leusen AM, Wynberg H (1983) Synthesis of enantiomerically pure (S)-(+)-3-hydroxytetrahydrofuran, and its (R)-enantiomer, from malic or tartaric acid. J Org Chem 48:2767–2769. doi:10.1021/jo00164a027
Valdehuesa KNG, Lee WK, Ramos KR, Cabulong RB, Choi J, Liu H, Nisola GM, Chung WJ (2015) Identification of aldehyde reductase catalyzing the terminal step for conversion of xylose to butanetriol in engineered Escherichia coli. Bioprocess Biosyst Eng. doi:10.1007/s00449-015-1417-4
Valdehuesa KNG, Liu H, Ramos KRM, Park SJ, Nisola GM, Lee WK, Chung WJ (2014) Direct bioconversion of d-xylose to 1,2,4-butanetriol in an engineered Escherichia coli. Process Biochem 49:25–32. doi:10.1016/j.procbio.2013.10.002
Van Dien S (2013) From the first drop to the first truckload: commercialization of microbial processes for renewable chemicals. Curr Opin Biotech 24:1061–1068. doi:10.1016/j.copbio.2013.03.002
Yim H, Haselbeck R, Niu W, Pujol-Baxley C, Burgard A, Boldt J, Khandurina J, Trawick JD, Osterhout RE, Stephen R, Estadilla J, Teisan S, Schreyer HB, Andrae S, Yang TH, Lee SY, Burk MJ, Van Dien S (2011) Metabolic engineering of Escherichia coli for direct production of 1,4-butanediol. Nat Chem Biol 7:445–452. doi:10.1038/Nchembio.580
Zhu L, Cai Z, Zhang Y, Li Y (2014) Engineering stress tolerance of Escherichia coli by stress-induced mutagenesis (SIM)-based adaptive evolution. Biotechnol J 9:120–127. doi:10.1002/biot.201300277
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This work was supported by the Knowledge Innovation Project of the Chinese Academy of Sciences (KSCX2-EW-G-5) and Beijing Municipal Natural Science Foundation (31170039).
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L. Sun and F. Yang have contributed equally to this work.
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Sun, L., Yang, F., Sun, H. et al. Synthetic pathway optimization for improved 1,2,4-butanetriol production. J Ind Microbiol Biotechnol 43, 67–78 (2016). https://doi.org/10.1007/s10295-015-1693-7
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DOI: https://doi.org/10.1007/s10295-015-1693-7