An Aspergillus awamori acetylesterase: purification of the enzyme, and cloning and sequencing of the gene.
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 alpha-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.
Koseki
T
T
National Research Institute of Brewing, Higashihiroshima, Japan.
Furuse
S
S
Iwano
K
K
Sakai
H
H
Matsuzawa
H
H
D000595:Amino Acid Sequence; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004950:Esterases / Q000096:biosynthesis; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000096:biosynthesis
D005656:Fungal Proteins; D004950:Esterases; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase; D016282:Aspartic Acid Endopeptidases; C026442:Aspergillus acid proteinase