Journal of Mushroom (한국버섯학회지)

The Korean Society of Mushroom Science (한국버섯학회)
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N-terminal amino acid sequence analysis of major manganese peroxidase (MnP3) produced by static culture of Pleurotus ostreatus

느타리 버섯균의 정치배양으로부터 생산되는 중요한 망간퍼옥시데이즈(MnP3)의 N-말단 아미노산 배열 분석

  • Ha, Hyo-Cheol (Department of bio-technology and convergence, Daegu Haany University)
  • 하효철 (대구한의대학교 바이오산업융합학부)
  • Received : 2019.08.28
  • Accepted : 2019.10.16
  • Published : 2019.12.31
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Abstract

Pleurotus ostreatus No.42, known as the ligninolytic basidiomycetes, showed production of MnP and Lac, but did not show any LiP acitivity in static culture, grown in GPYW liquid medium. Maximum production of MnP (80U/flask) was observed on day 11 of culturing in this medium. Chromatographic purification of MnP included the use of Sepharose CL-6B and Mono-Q. The major MnP isozyme purified by column chromatography was observed to be a 36.4 KDa (single band on SDS PAGE). The 19-amino acid sequence from the N-terminal was determined by protein sequencing to be ATCADGRTTANAACCVLFP. The N-terminal sequence of the major MnP isozyme of P. ostreatus No.42 was found to be the same as a previously reported sequence of an MnP3 isozyme from this fungus.

리그닌 분해 담자균류로 알려져 있는 느타리버섯균 No.42는 망간퍼옥시데이즈(MnP) 및 락게이즈(Lac)를 생산하였으나 글루코오스-펩톤-이스트-밀기울(GPYW)배지를 이용한 정치배양조건하에서 리그닌퍼옥시데이즈(LiP)활성은 검출되지 않았다. 한편, 동일배지에서 망간퍼옥시데이즈 활성은 11일째 80(3.6) U/flask(ml)의 최대 생산되었다. 망간퍼옥시데이즈 분리정제는 Sepha-ros CL-6B 및 Mono-Q 컬럼순으로 수행하였으며 주요 망간퍼옥시데이즈 isozyme은 단일밴드로 분자량은 36.4KDa이였다. N-말단으로부터 19개의 아미노산 배열은 단백질 자동 분석 장치로 분석한 결과 ATCADGRTTANAACCVLFP를 나타내었다. 느타리버섯균 No.42의 정치배양조건 하에서 세포외로 생산되는 중요한 망간퍼옥시데이즈 isozyme의 N-말단 아미노산 배열은 이전에 보고된 MnP3의 아미노산 배열과 동일하였다.

Keywords

References

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