caa a22 4500 605500010 CHVBK 20210128100553.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00253-015-6444-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6444-1 Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon [Elektronische Daten] [Seong Lee, Min-Sik Kim, Hae Jung, Jinwon Lee, Jung-Hyun Lee, Hyun Lee, Sung Kang] A strong promoter increases transcription of the genes of interest and enhances the production of various valuable substances. For a hyperthermophilic archaeon Thermococcus onnurineus NA1, which can produce H2 from carbon monoxide oxidation, we searched for a novel endogenous strong promoter by transcriptome analysis using high-throughput RNA sequencing. Based on the relative transcript abundance, we selected one promoter to encode a hypothetical gene, of which homologs were found only in several Thermococcales strains. This promoter, P TN0510 , was introduced into the front of CO-responsible hydrogenase gene cluster encoding a carbon monoxide dehydrogenase (CODH), a hydrogenase, and a Na+/H+ antiporter. In the resulting mutant strain, KS0510, transcription and translation level of the gene cluster increased by 4- to 14-folds and 1.5- to 1.9-folds, respectively, in comparison with those of the wild-type strain. Additionally, H2 production rate of KS0510 mutant was 4.8-fold higher than that of the wild-type strain. The P TN0510 was identified to be much stronger than the well-known two strong promoters, gdh and slp promoters from Thermococcus strains, through RT-qPCR and Western blotting analyses and kinetics of H2 production. In this study, we demonstrated that the RNA-seq approach is a good strategy to mine novel strong promoters of use to a Thermococcus strain when developed as a biotechnologically promising strain to produce valuable products such as enzymes and metabolites through metabolic engineering. Springer-Verlag Berlin Heidelberg, 2015 Strong promoter nationallicence RNA sequencing nationallicence Thermococcus onnurineus NA1 nationallicence Carbon monoxide nationallicence H2 production nationallicence Lee Seong Korea Institute of Ocean Science and Technology, Ansan, Korea aut Kim Min-Sik Korea Institute of Ocean Science and Technology, Ansan, Korea aut Jung Hae Korea Institute of Ocean Science and Technology, Ansan, Korea aut Lee Jinwon Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, Korea aut Lee Jung-Hyun Korea Institute of Ocean Science and Technology, Ansan, Korea aut Lee Hyun Korea Institute of Ocean Science and Technology, Ansan, Korea aut Kang Sung Korea Institute of Ocean Science and Technology, Ansan, Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/9(2015-05-01), 4085-4092 0175-7598 99:9<4085 2015 99 253 https://doi.org/10.1007/s00253-015-6444-1 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/s00253-015-6444-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lee Seong Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 700 1- Kim Min-Sik Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 700 1- Jung Hae Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 700 1- Lee Jinwon Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, Korea aut NATIONALLICENCE 700 1- Lee Jung-Hyun Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 700 1- Lee Hyun Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 700 1- Kang Sung Korea Institute of Ocean Science and Technology, Ansan, Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/9(2015-05-01), 4085-4092 0175-7598 99:9<4085 2015 99 253