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Isolation of a low-temperature adapted lipolytic enzyme from uncultivated microorganism
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- 발행기관
- 한국생물공학회 바로가기
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- 간행물
- 한국생물공학회 학술대회 바로가기
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- 통권
- 2012 춘계학술대회 및 국제심포지움 (2012.04)바로가기
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- 페이지
- pp.177-177
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- 저자
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- 언어
- 영어(ENG)
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- URL
- https://www.earticle.net/Article/A174448
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원문정보
초록
- 영어
- The aim of this study is to isolate a novel cold-adapted lipolytic enzyme from the activated sludge of uncultured microorganisms. The metagenomic DNA was directly extracted from the activated sludge, and a metagenomic library was constructed by using the pUC vector. The library was screened for lipolytic enzyme activity on 1% tributyrin agar plate. A clone among approximately 100,000 recombinant libraries showed lipolytic activity. The putative lipolytic gene encoding lipo1 from the metagenomic library was subcloned and expressed in Escherichia coli BL21 using the pET expression system. The expressed recombinant enzyme was purified by Ni-NTA affinity chromatography and characterized using general substrates of lipolytic property. The gene consisted of 972 bp encoding a polypeptide of 324 amino acids with a molecular mass of 35.6 kDa. Typical residues essential for lipolytic activity such as penta-peptide (GXSXG) and catalytic triad sequences (Ser166, Asp221 and His258) were detected. The deduced amino acid sequence of lipo1 exhibits low identity with amino acid sequences of esterase/lipase (32%, ZP_01528487) from Pseudomonas mendocina ymp and esterase (31%, AAY45707) from uncultured bacterium. This lipolytic enzyme exhibited the highest activity at pH 7.5 and 10°C. Thermal stability analysis showed that lipo1 was more unstable at 40°C than at 10°C. An activity-based strategy is an effective method for fishing out a low temperature-adapted lipolytic enzyme from the metagenomic library. This lipo1 enzyme can be considered to belong to the HSL family due to the enzyme’s oxyanion hole by the sequence HGGG. Lipo1 is a novel psychrophilic esterase obtained directly from the metagenomic library. Owing its support of significant activity at low temperature, this enzyme is expected to be useful for potential application as a biocatalyst in organic chemistry.
키워드
Metagenome Lipolytic enzyme Screening저자
간행물 정보
발행기관
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- 발행기관명
- 한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
- 설립연도
- 1984
- 분야
- 공학>생물공학
- 소개
- 이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다 1. 생물공학 분야의 발전을 위한 연구 협력 2. 생물공학의 실용화를 촉진시키기 위한 산학 협동 3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최 4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간 5. 생물공학 발전을 위한 정책 건의 6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동
간행물
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- 간행물명
- 한국생물공학회 학술대회
- 간기
- 반년간
- 수록기간
- 1985~2013
- 십진분류
- KDC 476 DDC 576
이 권호 내 다른 논문 / 한국생물공학회 학술대회 2012 춘계학술대회 및 국제심포지움
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