Comparison of CO-dependent H-2 production with strong promoters in Thermococcus onnurineus NA1
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Title
- Comparison of CO-dependent H-2 production with strong promoters in Thermococcus onnurineus NA1
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Author(s)
- Lee, Seong Hyuk; Kim, Min-Sik; Bae, Seung Seob; Choi, Ae Ran; Lee, Jin-Won; Kim, Tae Wan; 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)
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Alternative Author(s)
- 이성혁; 김민식; 배승섭; 최애란; 김태완; 이정현; 이현숙; 강성균
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Publication Year
- 2014-01
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Abstract
- To overproduce biotechnologically valuable products, the expression level of target genes has been modulated by using strong promoters. In a hyperthermophilic archaeon Thermococcus onnurineus NA1, two promoters, P (TN0413) and P (TN0157) , which drive expression of the genes encoding the S-layer protein and glutamate dehydrogenase were inserted in front of a gene cluster encoding a carbon monoxide dehydrogenase, a hydrogenase and a Na+/H+ antiporter. Two promoters exhibited strong activity by increasing the transcription and translation levels of the gene cluster in the mutant strains by 2.5- to 49-folds and 1.4- to 3.3-folds, respectively, than the native promoter in the wild-type strain. While KS0413 with P (TN0413) promoter exhibited 2.7 to 4.7 times higher transcript level than KS0157 with P (TN0157) promoter, the levels of proteins were a little different between them. The biomass concentrations and H-2 production rates of two mutants were 2- to 3-fold higher than those of the wild-type strain in a bioreactor where CO was supplied at a flow rate of 120 ml min(-1). Two mutants showed differential response to the higher CO flow rate, 240 ml min(-1), in terms of growth pattern and product formation, indicating two promoters were regulated by culture conditions. The results demonstrate that not only promoter strength but also product-forming conditions should be considered in promoter engineering.
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ISSN
- 0175-7598
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URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/2922
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DOI
- 10.1007/s00253-013-5448-y
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Bibliographic Citation
- APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.98, no.2, pp.979 - 986, 2014
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Publisher
- SPRINGER
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Subject
- SEA HYDROTHERMAL VENT; ESCHERICHIA-COLI; GENE-EXPRESSION; GLUTAMATE-DEHYDROGENASE; CONSTITUTIVE PROMOTERS; BACTERIA; CONSTRUCTION; SEQUENCE; ELEMENTS; CLONING
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Keywords
- Strong promoter; Hyperthermophilic archaeon; Carbon monoxide; H-2 production
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Type
- Article
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Language
- English
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Document Type
- Article
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