Screening of a novel strong promoter by RNA sequencing and its application to H-2 production in a hyperthermophilic archaeon SCIE SCOPUS

Cited 14 time in WEB OF SCIENCE Cited 14 time in Scopus
Title
Screening of a novel strong promoter by RNA sequencing and its application to H-2 production in a hyperthermophilic archaeon
Author(s)
Lee, Seong Hyuk; Kim, Min-Sik; Jung, Hae Chang; Lee, Jinwon; Lee, Jung-Hyun; Lee, Hyun Sook; Kang, Sung Gyun
KIOST Author(s)
Lee, Seong Hyuk(이성혁)Jung, Hae Chang(정해창)Lee, Jung Hyun(이정현)Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)
Publication Year
2015-05
Abstract
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 H-2 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, H-2 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 H-2 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.
ISSN
0175-7598
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/2488
DOI
10.1007/s00253-015-6444-1
Bibliographic Citation
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.99, no.9, pp.4085 - 4092, 2015
Publisher
SPRINGER
Subject
THERMOCOCCUS-ONNURINEUS NA1; CARBON-MONOXIDE; TRANSCRIPTION; GENERATION; DIVERSITY; REVEALS; FORMATE; SEQ
Keywords
Strong promoter; RNA sequencing; Thermococcus onnurineus NA1; Carbon monoxide; H-2 production
Type
Article
Language
English
Document Type
Article
Publisher
SPRINGER
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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