caa a22 4500 605505292 CHVBK 20210128100621.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6173-x doi (NATIONALLICENCE)springer-10.1007/s00253-014-6173-x Bacterial production of isobutanol without expensive reagents [Elektronische Daten] [Hironaga Akita, Nobutaka Nakashima, Tamotsu Hoshino] Isobutanol is attracting attention as a potential biofuel because it has higher energy density and lower hygroscopicity than ethanol. To date, several effective methods for microbial production of isobutanol have been developed, but they require expensive reagents to maintain expression plasmids and induce expression, which is not suitable for practical production. Here, we describe a simple and efficient method for isobutanol production in Escherichia coli. It is noteworthy that no expression plasmids or inducers were used during the production. Instead, heterologous genes necessary for isobutanol production were all knocked into the genome, and the expression of those genes was induced by xylose, which is present in most biomass feedstocks. The constructed strain (mlcXT7-LAFC-AAKCD) contains Bacillus subtilis alsS, E. coli ilvCD, Lactococcus lactis adhA, and L. lactis kivd genes in its genome and efficiently produced isobutanol from glucose and xylose in flask batch cultures. Under conditions in which the temperature and pH of the medium and the aeration in the culture were all optimized, the final isobutanol concentration reached 8.4gL−1 after 48h. Isobutanol was also produced using hydrolysate from Japanese cedar as the carbon source without supplemented glucose, xylose, or yeast extract. Under those conditions, isobutanol (3.7gL−1) was produced in 96h. Taken together, these results indicate that the developed strain is potentially useful for industrial isobutanol production. Springer-Verlag Berlin Heidelberg, 2014 BICES nationallicence Carbon catabolite repression nationallicence Isobutanol nationallicence Glucose nationallicence Lignocellulose biomass nationallicence Xylose nationallicence Akita Hironaga Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, 739-0046, Higashi-Hiroshima, Hiroshima, Japan aut Nakashima Nobutaka Bioproduction Research Institute, National Institute of Advanced Industrial Sciences and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, 062-8517, Sapporo, Toyohira-ku, Japan aut Hoshino Tamotsu Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, 739-0046, Higashi-Hiroshima, Hiroshima, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 991-999 0175-7598 99:2<991 2015 99 253 https://doi.org/10.1007/s00253-014-6173-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6173-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Akita Hironaga Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, 739-0046, Higashi-Hiroshima, Hiroshima, Japan aut NATIONALLICENCE 700 1- Nakashima Nobutaka Bioproduction Research Institute, National Institute of Advanced Industrial Sciences and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, 062-8517, Sapporo, Toyohira-ku, Japan aut NATIONALLICENCE 700 1- Hoshino Tamotsu Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, 739-0046, Higashi-Hiroshima, Hiroshima, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 991-999 0175-7598 99:2<991 2015 99 253