Enhanced production of 2,3-butanediol from xylose by combinatorial engineering of xylose metabolic pathway and cofactor regeneration in pyruvate decarboxylase-deficient Saccharomyces cerevisiae

• Published in: Bioresource technology 2017-12, Vol.245 (Pt B), p.1551-1557
• Main Authors: Kim, Soo-Jung, Sim, Hee-Jin, Kim, Jin-Woo, Lee, Ye-Gi, Park, Yong-Cheol, Seo, Jin-Ho
• Format: Article
• Language: English
• Subjects: 2,3-Butanediol; batch fermentation; biochemical pathways; Butylene Glycols; Candida tropicalis; Ethanol; Fermentation; genes; glycerol; Lactococcus lactis; Metabolic Engineering; Metabolic Networks and Pathways; noxE; oxidation; oxidoreductases; Pyruvate Decarboxylase; Pyruvate decarboxylase-deficient Saccharomyces cerevisiae; pyruvic acid; Saccharomyces cerevisiae; Scheffersomyces stipitis; TAL1; transaldolase; Xylose
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2017.06.034

2,3-Butanediol biosynthetic pathway from xylose and glucose in recombinant C2-independent and Pdc-deficient S. cerevisiae; The superscripts in the enzyme names present as follows: Bs, Bacillus subtilis; Ct, Candida tropicalis; Ll, Lactococcus lactis; Ss, Scheffersomyces stipitis; The names of enzymes are abbreviated as follow: XR, xylose reductase; XDH, xylitol dehydrogenase; XK, xylulose kinase; TAL1, transaldolase; ALS, α-acetolactate synthase; ALDC, α-acetolactate decarboxylase; BDH, 2,3-BDO dehydrogenase; PDC, pyruvate decarboxylase; NoxE, NADH oxidase. [Display omitted] •Overexpression of TAL1 improved 2,3-BDO productivity from xylose.•Expression of mutant XR contributed to enhancing xylose consumption rate.•Expression of noxE increased 2,3-BDO yield from xylose by reducing glycerol production.•Introduction of C. tropicalis PDC1 allowed to escape from C2-auxotrophy.•Optimal fed-batch fermentation resulted in 96.8g/L of 2,3-BDO from xylose. The aim of this study was to produce 2,3-butanediol (2,3-BDO) from xylose efficiently by modulation of the xylose metabolic pathway in engineered Saccharomyces cerevisiae. Expression of the Scheffersomyces stipitis transaldolase and NADH-preferring xylose reductase in S. cerevisiae improved xylose consumption rate by a 2.1-fold and 2,3-BDO productivity by a 1.8-fold. Expression of the Lactococcus lactis noxE gene encoding NADH oxidase also increased 2,3-BDO yield by decreasing glycerol accumulation. Additionally, the disadvantage of C2-dependent growth of pyruvate decarboxylase-deficient (Pdc−) S. cerevisiae was overcome by expression of the Candida tropicalis PDC1 gene. A fed-batch fermentation of the BD5X-TXmNP strain resulted in 96.8g/L 2,3-BDO and 0.58g/L-h productivity from xylose, which were 15.6- and 2-fold increases compared with the corresponding values of the BD5X strain. It was concluded that facilitation of the xylose metabolic pathway, oxidation of NADH and relief of C2-dependency synergistically triggered 2,3-BDO production from xylose in Pdc−S. cerevisiae.