Screening of a novel strong promoter by RNA sequencing and its application to H-2 production in a hyperthermophilic archaeon
SCIE
SCOPUS
Cited 20 time in
WEB OF SCIENCE
Cited 21 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); Lee, Jung Hyun(이정현); Lee, Hyun Sook(이현숙); Kang, Sung Gyun(강성균)
-
Alternative Author(s)
- 이성혁; 김민식; 정해창; 이정현; 이현숙; 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
- Files in This Item:
-
There are no files associated with this item.
Items in ScienceWatch@KIOST are protected by copyright, with all rights reserved, unless otherwise indicated.