An inducible acetylesterase was purified from the culture medium of Aspergillus awamori strain IFO4033 growing on wheat-bran culture by ion-exchange, gel-filtration and hydrophobic-interaction chromatographies. The purified enzyme had an Mr of 31000 and contained Asn-linked oligosaccharides. The enzyme liberated acetic acid from wheat bran, hydrolysed only α-naphthyl acetate and propionate when aromatic esters were used for the substrate, and was tentatively classified as a carboxylic esterase (EC 3.1.1.1). The gene encoding acetylesterase was cloned and sequenced. The deduced amino acid sequence showed that acetylesterase was produced as a 304-amino-acid-residue precursor, which was converted post-translationally into a 275-amino-acid-residue mature protein. Part of the sequence of acetylesterase was similar to the region near the active-site serine of lipases of Geotrichum candidum and Candida cylindracea. A unique site of putative Asn-linked oligosaccharides was presented.
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September 1997
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Research Article|
September 01 1997
An Aspergillus awamori acetylesterase: purification of the enzyme, and cloning and sequencing of the gene
Takuya KOSEKI;
*National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashihiroshima 739, Japan
2To whom correspondence should be addressed.
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Shinji FURUSE;
Shinji FURUSE
†General Research Laboratory, Ozeki Corporation, 4-9 Imazu Dezaike-cho, Nishinomiya, Hyogo 663, Japan
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Kimio IWANO;
Kimio IWANO
*National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashihiroshima 739, Japan
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Hiroshi SAKAI;
Hiroshi SAKAI
‡Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
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Hiroshi MATSUZAWA
Hiroshi MATSUZAWA
‡Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan
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Publisher: Portland Press Ltd
Received:
December 24 1996
Revision Received:
April 24 1997
Accepted:
May 01 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 326 (2): 485–490.
Article history
Received:
December 24 1996
Revision Received:
April 24 1997
Accepted:
May 01 1997
Citation
Takuya KOSEKI, Shinji FURUSE, Kimio IWANO, Hiroshi SAKAI, Hiroshi MATSUZAWA; An Aspergillus awamori acetylesterase: purification of the enzyme, and cloning and sequencing of the gene. Biochem J 1 September 1997; 326 (2): 485–490. doi: https://doi.org/10.1042/bj3260485
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