TAILIEUCHUNG - Kataoka et al. AMB Express 2011, 1:10 http://www.amb-express.com/content/1/1/10 ORIGINAL Open

Kataoka et al. AMB Express 2011, 1:10 ORIGINAL Open Access Development of butanol-tolerant Bacillus subtilis strain GRSW2-B1 as a potential bioproduction host Naoya Kataoka1, Takahisa Tajima1, Junichi Kato1, Wanitcha Rachadech2 and Alisa S Vangnai 2,3* Abstract As alternative microbial hosts for butanol production with organic-solvent tolerant trait are in high demands, a butanol-tolerant bacterium, Bacillus subtilis GRSW2-B1, was thus isolated. Its tolerance covered a range of organic solvents at high concentration (5%v/v), with remarkable tolerance in particular to butanol and alcohol groups. It was susceptible for butanol acclimatization, which resulted in significant tolerance improvement. It has versatility for application in a variety of fermentation process. | Kataoka et al. AMB Express 2011 1 10 http content 1 1 10 o AMB Express a SpringerOpen Journal ORIGINAL Open Access Development of butanol-tolerant Bacillus subtilis strain GRSW2-B1 as a potential bioproduction host Naoya Kataoka1 Takahisa Tajima1 Junichi Kato1 Wanitcha Rachadech2 and Alisa S Vangnai 2 3 Abstract As alternative microbial hosts for butanol production with organic-solvent tolerant trait are in high demands a butanol-tolerant bacterium Bacillus subtilis GRSW2-B1 was thus isolated. Its tolerance covered a range of organic solvents at high concentration 5 v v with remarkable tolerance in particular to butanol and alcohol groups. It was susceptible for butanol acclimatization which resulted in significant tolerance improvement. It has versatility for application in a variety offermentation process because it has superior tolerance when cells were exposed to butanol either as high-density late-exponential grown cells up to 5 v v or under growing conditions up to v v . Genetic transformation procedure was optimized yielding the highest efficiency at X 103 colony forming unit pg DNA -1. Gene expression could be effectively driven by several promoters with different levels where as the highest expression was observed with a xylose promoter. The constructed vector was stably maintained in the transformants in the presence or absence of butanol stress. Adverse effect of efflux-mediated tetracycline resistance determinant TetL to bacterial organic-solvent tolerance property was unexpectedly observed and thus discussed. Overall results indicate that B. subtilis GRSW2-B1 has potential to be engineered and further established as a genetic host for bioproduction of butanol. Keywords Organic-solvent tolerant bacteria Butanol-tolerant bacteria Heterologous gene-expression host Introduction n-Butanol hereafter referred to as butanol is an important industrial chemical widely used as a solvent a stabilizer and feedstock for the production .

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