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논문 기본 정보

자료유형
학술저널
저자정보
Yao Zhuang (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) Meng Yu (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) Le Huong Giang (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) Lee Se Jin (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) Jeon Hye Sung (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) Ji Yeon Yoo (Division of Applied Life Science (BK21 Four) Graduate School Gyeongsang National University Jinju 5) 김정환 (경상대학교)
저널정보
한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제31권 제6호
발행연도
2021.1
수록면
833 - 839 (7page)

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Bacillus subtilis CH3-5 isolated from cheonggukjang secretes a 28 kDa protease with a strong fibrinolytic activity. Its gene, aprE3-5, was cloned and expressed in a heterologous host (Jeong et al., 2007). In this study, the promoter of aprE3-5 was replaced with other stronger promoters (Pcry3A, P10, PSG1, PsrfA) of Bacillus spp. using PCR. The constructed chimeric genes were cloned into pHY300PLK vector, and then introduced into B. subtilis WB600. The P10 promoter conferred the highest fibrinolytic activity, i.e., 1.7-fold higher than that conferred by the original promoter. Overproduction of the 28 kDa protease was confirmed using SDS-PAGE and fibrin zymography. RTqPCR analysis showed that aprE3-5 expression was 2.0-fold higher with the P10 promoter than with the original promoter. Change of the initiation codon from GTG to ATG further increased the fibrinolytic activity. The highest aprE3-5 expression was observed when two copies of the P10 promoter were placed in tandem upstream of the ATG initiation codon. The construct with P10 promoter and ATG and the construct with two copies of P10 promoter in tandem and ATG exhibited 117% and 148% higher fibrinolytic activity, respectively, than that exhibited by the construct containing P10 promoter and GTG. These results confirmed that significant overproduction of a fibrinolytic enzyme can be achieved by suitable promoter modification, and this approach may have applications in the industrial production of AprE3-5 and related fibrinolytic enzymes.

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