Xylose isomerase overexpression along with engineering of the pentose phosphate pathway and evolutionary engineering enable rapid xylose utilization and ethanol production by Saccharomyces cerevisiae

Zhou, Hang; Cheng, Jun; Wang, Benjamin; Fink, Gerald R.; Stephanopoulos, Gregory

doi:10.1016/j.ymben.2012.07.011

Cited by 264 publications

(268 citation statements)

References 41 publications

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“…Here we engineered a new pathway not present in nature into yeast to couple xylose fermentation and acetate detoxification under industrially relevant anaerobic conditions. Although the XR/XDH pathway usually generates surplus NADH compared with the alternative redox-neutral XI pathway 34 and results in wasteful byproducts, we show that the surplus NADH can be used as an advantage to drive reduction of acetate into ethanol. Meanwhile, the ATP need for activating acetate to acetyl-CoA before AADH reaction may be provided by the co-fermentation of xylose.…”

Section: Discussionmentioning

confidence: 86%

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

et al. 2013

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The anticipation for substituting conventional fossil fuels with cellulosic biofuels is growing in the face of increasing demand for energy and rising concerns of greenhouse gas emissions. However, commercial production of cellulosic biofuel has been hampered by inefficient fermentation of xylose and the toxicity of acetic acid, which constitute substantial portions of cellulosic biomass. Here we use a redox balancing strategy to enable efficient xylose fermentation and simultaneous in situ detoxification of cellulosic feedstocks. By combining a nicotinamide adenine dinucleotide (NADH)-consuming acetate consumption pathway and an NADH-producing xylose utilization pathway, engineered yeast converts cellulosic sugars and toxic levels of acetate together into ethanol under anaerobic conditions. The results demonstrate a breakthrough in making efficient use of carbon compounds in cellulosic biomass and present an innovative strategy for metabolic engineering whereby an undesirable redox state can be exploited to drive desirable metabolic reactions, even improving productivity and yield.

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Section: Discussionmentioning

confidence: 86%

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

et al. 2013

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“…Various attempts have also been made to establish xylose metabolic pathway into S. cerevisiae through genetic engineering by incorporating xylose reductase-xylitol dehydrogenase or xylose isomerase pathway for the coutilization of glucose and xylose (Kuyper et al, 2005;Liu and Hu, 2010;Zhou et al, 2012). However, there are various limitations including competitive inhibition due to presence of glucose and degradation of xylose transporter, which inhibits coutilization of xylose (Apel et al, 2016).…”

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confidence: 99%

“…Evolutionary adaptation together with genetic engineering can be a better approach to develop the sustainable and economic process (Behera et al, 2016;Kuyper et al, 2005;Liu and Hu, 2010;Zhou et al, 2012;Sharma et al, 2016). This strategy has already been implemented along with various conventional techniques including protein engineering, intergeneric hybridization, and metabolic engineering (Kuhad et al, 2011;Zhang et al, 2015).…”

mentioning

confidence: 99%

“…This strategy has already been implemented along with various conventional techniques including protein engineering, intergeneric hybridization, and metabolic engineering (Kuhad et al, 2011;Zhang et al, 2015). Furthermore, evolutionary adaptation has been proven a useful approach for the development of sustainable technology (Kuyper et al, 2005;Liu and Hu, 2010;Zhou et al, 2012;Pereira et al, 2015).…”

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confidence: 99%

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Evolutionary Adaptation of Kluyveromyces marxianus NIRE-K3 for Enhanced Xylose Utilization

et al. 2017

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The evolutionary adaptation was approached on the thermotolerant yeast Kluyveromyces marxianus NIRE-K3 at 45°C on xylose as a sole source of carbon for enhancement of xylose uptake. After 60 cycles, evolved strain K. marxianus NIRE-K3.1 showed comparatively 3.75-and 3.0-fold higher specific growth and xylose uptake rates, respectively, than that of native strain. Moreover, the short lag phase was also observed on adapted strain. During batch fermentation with xylose concentration of 30 g l −1, K. marxianus NIRE-K3.1 could utilize about 96% of xylose in 72 h and produced 4.67 and 15.7 g l −1 of ethanol and xylitol, respectively, which were 9.72-and 4.63-fold higher than that of native strain. Similarly, specific sugar consumption rate, xylitol, and ethanol yields were 5.07-, 1.15-, and 2.44-fold higher as compared to the native strain, respectively. The results obtained after evolutionary adaptation of K. marxianus NIRE-K3 show the significant improvement in the xylose utilization, ethanol and xylitol yields, and productivities. By understanding the results obtained, the significance of evolutionary adaptation has been rationalized, since the adapted culture could be more stable and could enhance the productivity.

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“…Two types of xylose-assimilating pathways have been identified and used to engineer xylose-utilizing S. cerevisiae strains. One is the redox cofactor-dependent xylose reductase (XR)/xylitol dehydrogenase (XDH) pathway (8)(9)(10)(11)(12)(13)(14)(15), and the other is the redoxneutral xylose isomerase (XI) pathway (16)(17)(18)(19)(20)(21)(22)(23). Introduction of the XR/XDH pathway into S. cerevisiae by metabolic engineering approaches has been widely studied, but redox imbalance is a key problem, because XR can use both NADPH and NADH, while XDH uses NAD ϩ exclusively (6,24).…”

mentioning

confidence: 99%

Deletion of FPS1 , Encoding Aquaglyceroporin Fps1p, Improves Xylose Fermentation by Engineered Saccharomyces cerevisiae

Wei

Kim

et al. 2013

Appl Environ Microbiol

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Accumulation of xylitol in xylose fermentation with engineered Saccharomyces cerevisiae presents a major problem that hampers economically feasible production of biofuels from cellulosic plant biomass. In particular, substantial production of xylitol due to unbalanced redox cofactor usage by xylose reductase (XR) and xylitol dehydrogenase (XDH) leads to low yields of ethanol. While previous research focused on manipulating intracellular enzymatic reactions to improve xylose metabolism, this study demonstrated a new strategy to reduce xylitol formation and increase carbon flux toward target products by controlling the process of xylitol secretion. Using xylitol-producing S. cerevisiae strains expressing XR only, we determined the role of aquaglyceroporin Fps1p in xylitol export by characterizing extracellular and intracellular xylitol. In addition, when FPS1 was deleted in a poorly xylose-fermenting strain with unbalanced XR and XDH activities, the xylitol yield was decreased by 71% and the ethanol yield was substantially increased by nearly four times. Experiments with our optimized xylose-fermenting strain also showed that FPS1 deletion reduced xylitol production by 21% to 30% and increased ethanol yields by 3% to 10% under various fermentation conditions. Deletion of FPS1 decreased the xylose consumption rate under anaerobic conditions, but the effect was not significant in fermentation at high cell density. Deletion of FPS1 resulted in higher intracellular xylitol concentrations but did not significantly change the intracellular NAD ؉ / NADH ratio in xylose-fermenting strains. The results demonstrate that Fps1p is involved in xylitol export in S. cerevisiae and present a new gene deletion target, FPS1, and a mechanism different from those previously reported to engineer yeast for improved xylose fermentation.

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Xylose isomerase overexpression along with engineering of the pentose phosphate pathway and evolutionary engineering enable rapid xylose utilization and ethanol production by Saccharomyces cerevisiae

Cited by 264 publications

References 41 publications

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

Evolutionary Adaptation of Kluyveromyces marxianus NIRE-K3 for Enhanced Xylose Utilization

Deletion of FPS1 , Encoding Aquaglyceroporin Fps1p, Improves Xylose Fermentation by Engineered Saccharomyces cerevisiae

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