Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae

Zhang, Yanfei; Lane, Stephan; Chen, Jhong-Min; Hammer, Sarah K.; Luttinger, Jake; Yang, Lifeng; Jin, Yong-Su; Avalos‬, José L.

doi:10.1186/s13068-019-1560-2

Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae

Journal Article · Fri Sep 20 00:00:00 EDT 2019 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-019-1560-2· OSTI ID:1618761

Zhang, Yanfei; Lane, Stephan; Chen, Jhong-Min; Hammer, Sarah K.; Luttinger, Jake; Yang, Lifeng; Jin, Yong-Su;

Abstract Background

Branched-chain higher alcohols (BCHAs), including isobutanol and 2-methyl-1-butanol, are promising advanced biofuels, superior to ethanol due to their higher energy density and better compatibility with existing gasoline infrastructure. Compartmentalizing the isobutanol biosynthetic pathway in yeast mitochondria is an effective way to produce BCHAs from glucose. However, to improve the sustainability of biofuel production, there is great interest in developing strains and processes to utilize lignocellulosic biomass, including its hemicellulose component, which is mostly composed of the pentose xylose.

Results

In this work, we rewired the xylose isomerase assimilation and mitochondrial isobutanol production pathways in the budding yeast Saccharomyces cerevisiae . We then increased the flux through these pathways by making gene deletions of BAT1 , ALD6 , and PHO13 , to develop a strain (YZy197) that produces as much as 4 g/L of BCHAs (3.10 ± 0.18 g isobutanol/L and 0.91 ± 0.02 g 2-methyl-1-butanol/L) from xylose. This represents approximately a 28-fold improvement on the highest isobutanol titers obtained from xylose previously reported in yeast and the first report of 2-methyl-1-butanol produced from xylose. The yield of total BCHAs is 57.2 ± 5.2 mg/g xylose, corresponding to ~ 14% of the maximum theoretical yield. Respirometry experiments show that xylose increases mitochondrial activity by as much as 7.3-fold compared to glucose.

Conclusions

The enhanced levels of mitochondrial BCHA production achieved, even without disrupting ethanol byproduct formation, arise mostly from xylose activation of mitochondrial activity and are correlated with slow rates of sugar consumption.

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Research Organization:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: SC0018420; SC0019363

OSTI ID:: 1618761

Alternate ID(s):: OSTI ID: 1571260

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 12; ISSN 1754-6834

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: Netherlands

Language:: English

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09 BIOMASS FUELS
2-methyl-1-butanol
Saccharomyces cerevisiae
branched-chain higher alcohols
isobutanol
mitochondrial engineering
xylose

Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae

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