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

고초균 아종 spizizenii의 α-acetolactate decarboxylase 결정 구조

  • 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)
  • 엄지영 (강원대학교 의생명과학대학 의생명융합학부) ;
  • 오한별 (강원대학교 의생명과학대학 의생명융합학부) ;
  • 윤성일 (강원대학교 의생명과학대학 의생명융합학부)
  • Received : 2019.01.07
  • Accepted : 2019.01.29
  • Published : 2019.03.31

Abstract

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.

Keywords

Bacillus subtilis;acetoin;${\alpha}$-acetolactate decarboxylase;structure;zinc

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

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

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Fig. 3. Overall structure of bssALDC monomer.

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Fig. 4. bssALDC dimer and its dimerization interface.

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

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Fig. 6. Zinc binding of bssALDC.

Table 1. Crystallographic statistics of the bssALDC structure

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Acknowledgement

Supported by : National Research Foundation of Korea, Kangwon National University

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