Comparison of CO-dependent H2 Production with Strong Promoters in Thermococcus onnurineus NA1

Title
Comparison of CO-dependent H2 Production with Strong Promoters in Thermococcus onnurineus NA1
Author(s)
이성혁; 김민식; 배승섭; 최애란; 이진원; 김태완; 이정현; 이현숙; 강성균
KIOST Author(s)
Lee, Seong Hyuk(이성혁)Lee, Jung Hyun(이정현)Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)
Publication Year
2014-05-02
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, PTN0413 and PTN0157, which drive expression of the genes encoding the Slayer 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 PTN0413 promoter exhibited 2.7 to 4.7 times higher transcript level than KS0157 with PTN0157 promoter, the levels of proteins were a little different between them. The biomass concentrations and H2 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.rive expression of the genes encoding the Slayer 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 PTN0413 promoter exhibited 2.7 to 4.7 times higher transcript level than KS0157 with PTN0157 promoter, the levels of proteins were a little different between them. The biomass concentrations and H2 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.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/26296
Bibliographic Citation
한국미생물학회 국제학술대회, pp.151, 2014
Publisher
한국미생물학회
Type
Conference
Language
English
Publisher
한국미생물학회
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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