Korean Journal of Microbiology (미생물학회지)
- Volume 32 Issue 3
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- Pages.215-221
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- 1994
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- 0440-2413(pISSN)
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- 2383-9902(eISSN)
Nucleotide Sequence of the Penicillin G Acylase Gene from Bacillus megaterium and Characteristics of the Enzyme
Bacillus megaterium에서 발견된 Penicillin G Acylse 유전자의 염기서열과 그 효소의 특성
- Gang, Ju-Hyeon (Department of Life Science, Korea Advanced Institute of Science and Technology) ;
- Kim, Seong-Jae (Department of Microbiology, Chungbuk National University) ;
- Park, Yong-Chjun (Department of Microbiology, Chungbuk National University) ;
- Hwang, Young (Department of Life Science, Korea Advanced Institute of Science and Technology) ;
- Yoo, Ook-Joon (Department of Life Science, Korea Advanced Institute of Science and Technology) ;
- Kim, Young-Chang (Department of Microbiology, Chungbuk National University)
- 강주현 (한국과학기술원 생명과학과) ;
- 김성재 (충북대학교 자연과학대학 미생물학과) ;
- 박용춘 (충북대학교 자연과학대학 미생물학과) ;
- 황영 (한국과학기술원 생명과학과) ;
- 유욱준 (한국과학기술원 생명과학과) ;
- 김영창 (충북대학교 자연과학대학 미생물학과)
- Published : 1994.01.01
Abstract
The complete nucleotide sequence of the cloned pga gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 and its 5'- and 3'-flanking regions was determined. The sequence revealed only one large open reading frame (2,406 hp) of the penicillin G acylase (pga) gene. Upstream from ATG of the pga gene, there was a putative ribosome binding site, Shine-Dalgarno sequence. The promoter-like structure, - 10 and - 35 sequences, was also found. Following the stop codon, TAG, a structure reminiscent of the E. coli rho-independent transcription terminator was present. The amino acid sequence was deduced from the nucleotide sequence. The molecular mass of the polypeptide was 91,983 Da. There was a potential signal sequence in its amino-terminal region. A comparison of its deduced amino acid sequence with other characterized penicillin G acylases and the result of SDS-polyacrylamide gel electrophoresis of the purified enzyme showed that a precursor polypeptide of 92 kDa was processed into two dissimilar
Bacillus megaterium ATCC 14945의 penicillin G acylase 유전자의 염기배열을 결정하였다. 이 유전자에는 2,406 염기쌍으로 이루어진 하나의 open reading frame이 존재하는데, 개시코돈의 5' 위쪽에서 Shine-Dalgarno 배열과 promoter로 여겨지는 부분을 발견하였으며, 종결코돈의 3' 아래쪽에서 rho-independent한 전사종결체와 dby사한 구조를 발견하였다. 염기배열로부터 폴리펩티드의 아미노산 배열을 유추하였다. 이 폴리펩티드의 분자량은 91,983 Da이었으며, 아미노 말단 부이에 signal sequence가 존재하였다. 이 아미노산 배열을 여러 다른 penicillin G acylase의 아미노산 배열과 비교하고 분리 정제한 효소를 SDS-polyacrylamide gel 전기영동으로 분석한 결과로부터 이 효소는 92kDa의 전구체로 해독된 후 processing 과정을 거쳐 각각 25kDa과 61kDa의