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Title 

Enhancing the 1-butanol tolerance in Escherichia coli through repetitive proton beam irradiation

Authors 

Haeyoung JeongJ Han

Publisher 

Korean Physical Society

Issue Date 

2010

Citation 

Journal of the Korean Physical Society, vol. 56, no. 6, pp. 2041-2045

Keywords 

1-butanolEscherichia coliGenome sequencingMutationProton beamTolerance

Abstract 

The low butanol tolerance of most microorganisms severely limits the production of 1-butanol on an economical scale in alternative hosts other than the natural butanol producer Clostridium acetobutylicum, which is not amenable for genetic manipulation and requires demanding culture conditions. To generate butanol-tolerant E. coli, we devised a cyclic selection strategy that consists of an iterative application of proton irradiation at a dose of ∼250 Gy by using 45-MeV protons and selection by daily serial transfer in a minimal medium containing increasing concentrations of 1-butanol. Applying five rounds of the cyclic selection of E. coli ATCC 8739 (a C strain) over 61 days resulted in a mutant population that could tolerate 1.2% 1-butanol (v/v). However, without proton irradiation, the cells were unable to grow at ≥0.8% 1-butanol in a control experiment. Seven different mutations were identified within one clone from the endpoint population through 454 pyrosequencing of the genome. Tracing each mutation in terms of the prevalence in the population during the period of evolution suggested that proton beam irradiation-induced mutations were rapidly fixed during the early phase of the selection procedure. This approach, which is still being applied to increase butanol tolerance beyond 1.3%, can be considered useful for improving targeted traits whose corresponding genes are unknown.

ISSN 

0374-4884

Link 

http://dx.doi.org/10.3938/jkps.56.2041

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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