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  • Inverse metabolic engineering of Bacillus subtilis for xylose utilization based on adaptive evolution and whole-genome sequencing.

Inverse metabolic engineering of Bacillus subtilis for xylose utilization based on adaptive evolution and whole-genome sequencing.

Applied microbiology and biotechnology (2015-01-27)
Bo Zhang, Ning Li, Zhiwen Wang, Ya-Jie Tang, Tao Chen, Xueming Zhao
ABSTRACT

Efficient utilization of xylose by bacteria is essential for production of fuels and chemicals from lignocellulosic biomass. In this study, Bacillus subtilis 168 was subjected to laboratory adaptive evolution, and a mutant E72, which could grow on xylose with a maximum specific growth rate of 0.445 h(-1), was obtained. By whole-genome sequencing, 16 mutations were identified in strain E72. Through further analysis, three of them, which were in the coding regions of genes araR, sinR, and comP, were identified as the beneficial mutations. The reconstructed strain 168ARSRCP harboring these three mutations exhibited similar growth capacity on xylose to the evolved strain E72, and the average xylose consumption rate of this strain is 0.530 g/l/h, much higher than that of E72 (0.392 g/l/h). Furthermore, genes acoA and bdhA were deleted and the final strain could utilize xylose to produce acetoin at 71 % of the maximum theoretical yield. These results suggested that this strain could be used as a potential platform for production of fuels and chemicals from lignocellulosic biomass.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetoin, May exist as crystalline dimer
Pricing and availability is not currently available.
Sigma-Aldrich
Acetoin, primarily dimer, ≥95%, FG
Pricing and availability is not currently available.
Supelco
Acetoin, analytical standard
Pricing and availability is not currently available.
Sigma-Aldrich
Acetoin, natural, ≥95%, FG
Pricing and availability is not currently available.
Sigma-Aldrich
2,3-Butanediol, 98%
Pricing and availability is not currently available.
Supelco
2,3-Butanediol, analytical standard, mixture of racemic and meso forms
Pricing and availability is not currently available.
Xylose, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.
Sigma-Aldrich
D-(+)-Xylose, ≥99%
Pricing and availability is not currently available.
Sigma-Aldrich
D-(+)-Xylose, ≥99% (GC)
Pricing and availability is not currently available.
Sigma-Aldrich
D-(+)-Xylose, BioXtra, ≥99% (GC)
Pricing and availability is not currently available.
Sigma-Aldrich
D-(+)-Xylose, BioUltra, ≥99.0% (sum of enantiomers, HPLC)
Pricing and availability is not currently available.