미생물학회지 (Korean Journal of Microbiology)

  • 제55권1호
  • /
  • Pages.9-16
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  • 2019
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
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고초균 아종 spizizenii의 α-acetolactate decarboxylase 결정 구조

Crystal structure of α-acetolactate decarboxylase from Bacillus subtilis subspecies spizizenii

  • 엄지영 (강원대학교 의생명과학대학 의생명융합학부) ;
  • 오한별 (강원대학교 의생명과학대학 의생명융합학부) ;
  • 윤성일 (강원대학교 의생명과학대학 의생명융합학부)
  • Eom, Jiyoung (Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University) ;
  • Oh, Han Byeol (Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University) ;
  • Yoon, Sung-il (Division of Biomedical Convergence, College of Biomedical Science, Kangwon National University)
  • 투고 : 2019.01.07
  • 심사 : 2019.01.29
  • 발행 : 2019.03.31
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초록

다양한 미생물은 세포와 주변의 과산화를 방지하고 여분의 에너지를 보관하기 위해 ${\alpha}$-acetolactate decarboxylase(ALDC)를 이용해 아세토인을 생성한다. 아세토인은 안전한 식품 향미 개선제이기 때문에 ALDC를 이용한 아세토인 생합성에 많은 산업체가 관심을 가지고 있다. ALDC는 ${\alpha}$-acetolactate의 탈카르복실화 반응을 통해 아세토인을 생산하는 금속 의존 효소이다. 본 논문에서는 고초균 아종 spizizenii의 ALDC(bssALDC) 결정구조를 $1.7{\AA}$ 해상도에서 보고한다. bssALDC는 두 개의 ${\beta}$-sheet가 중앙부를 형성하는 ${\alpha}/{\beta}$ 구조를 가진다. bssALDC는 중앙부의 소수성 상호작용과 주변부의 친수성 상호작용을 통해 이합체를 형성한다. bssALDC는 세 개의 histidine 잔기와 세 개의 물 분자를 이용해 아연 이온에 배위결합한다. 구조와 서열의 비교 분석에 기초하여 아연 이온과 이 주변부 bssALDC 잔기들이 bssALDC의 효소 활성부위임을 제안한다.

Acetoin is generated by numerous microorganisms using ${\alpha}$-acetolactate decarboxylase (ALDC) to prevent overacidification of cells and their environment and to store remaining energy. Because acetoin has been used as a safe flavor enhancer in food products, industries have been interested in biotechnological production of acetoin using ALDC. ALDC is a metal-dependent enzyme that produces acetoin from ${\alpha}$-acetolactate through decarboxylation reaction. Here, we report the crystal structure of ALDC from Bacillus subtilis subspecies spizizenii (bssALDC) at $1.7{\AA}$ resolution. bssALDC folds into a two-domain ${\alpha}/{\beta}$ structure where two ${\beta}$-sheets form a central core. bssALDC assembles into a dimer through central hydrophobic interactions and peripheral hydrophilic interactions. bssALDC coordinates a zinc ion using three histidine residues and three water molecules. Based on comparative analyses of ALDC structures and sequences, we propose that the active site of bssALDC includes the zinc ion and its neighboring bssALDC residues.

키워드

MSMHBQ_2019_v55n1_9_f0001.png 이미지

Fig. 1. SDS-PAGE analysis of purified bssALDC protein.

MSMHBQ_2019_v55n1_9_f0002.png 이미지

Fig. 2. Amino acid sequence alignment of bssALDC and its orthologs (bs168ALDC, bbALDC, and kpALDC).

MSMHBQ_2019_v55n1_9_f0003.png 이미지

Fig. 3. Overall structure of bssALDC monomer.

MSMHBQ_2019_v55n1_9_f0004.png 이미지

Fig. 4. bssALDC dimer and its dimerization interface.

MSMHBQ_2019_v55n1_9_f0005.png 이미지

Fig. 5. Dimerization interface of bssALDC and its sequence conservation.

MSMHBQ_2019_v55n1_9_f0006.png 이미지

Fig. 6. Zinc binding of bssALDC.

Table 1. Crystallographic statistics of the bssALDC structure

MSMHBQ_2019_v55n1_9_t0001.png 이미지

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