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Engineering of Klebsiella oxytoca for the Production of 2,3-Butanediol from High Concentration of Xylose

Authors
Cha, Ji WonJang, Seung HoonSon, JaewooKim, JungyeonJung, InhoKim, Kyoung HeonChang, Yong KeunJeong, Ki Jun
Issue Date
1-11월-2021
Publisher
AMER CHEMICAL SOC
Keywords
2,3-butanediol; Klebsiella oxytoca; adaptive laboratory evolution; biosensor; xylose
Citation
ACS SUSTAINABLE CHEMISTRY & ENGINEERING, v.9, no.43, pp.14395 - 14404
Indexed
SCIE
SCOPUS
Journal Title
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
Volume
9
Number
43
Start Page
14395
End Page
14404
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/135797
DOI
10.1021/acssuschemeng.1c04118
ISSN
2168-0485
Abstract
Klebsiella oxytoca is widely used for the biological production of 2,3-butanediol (2,3-BDO), a promising platform chemical with a broad range of applications. Here, to improve cell growth and production of 2,3-BDO under high concentration of xylose (100 g/L), we engineered K. oxytoca using an adaptive laboratory evolution and a biosensor-derived high throughput screening strategy. First, we developed a XylR-dependent xylose biosensor for the detection of intracellular xylose, and K. oxytoca containing the xylose biosensor was used for adaptive laboratory evolution in 100 g/L xylose. Cells were isolated by FACS screening, and the isolated strain (KO8S16) showed much improved cell growth with high xylose consumption rate (1.35 g/L/h) and 2,3-BDO productivity (0.53 g/L/h) compared with the wild-type strain. Through whole genome resequencing, it was revealed that a mutation in OmpR (a response regulator of osmotic stress) allowed to withstand high concentrations of xylose. Finally, fed-batch cultivation was performed by feeding high concentration of xylose, and K. oxytoca successfully produced 2,3-BDO at a concentration as high as 57.5 g/L by consuming 238.13 g/L xylose in 47 h.
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