High Recovery Purification and Some Properties of a<i>β</i>-Glucosidase from<i>Aspergillus niger</i>

Unno, Takehiro; Ide, Katsutoshi; Yazaki, Terutaka; Tanaka, Yoshimasa; Nakakuki, Teruo; Okada, Gentaro

doi:10.1271/bbb.57.2172

Cited by 34 publications

(13 citation statements)

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“…This enzyme effectively hydrolyzes flavor compound glycosides in certain low pH products, such as wine and passion fruit juice, thereby enhancing their flavor (8 -11) and is particularly attractive for use in the food industry because A. niger is considered nontoxic (3). Other A. niger ␤-glucosidases have also been purified (12)(13)(14); however, differences in their properties have been reported, including ranges of molecular masses (116 -137 kDa) and isoelectric points (pI values of 3.8 -4) and pH optima (3.4 -4.5). Indeed, at least two ␤-glucosidases with distinct substrate specificities have been identified in commercial A. niger ␤-glucosidase preparations (15).…”

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confidence: 99%

Cloning, Expression, Characterization, and Nucleophile Identification of Family 3, Aspergillus nigerβ-Glucosidase

Dan¹,

Marton²,

Dekel³

et al. 2000

Journal of Biological Chemistry

125

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The ␤-glucosidase from Aspergillus niger (CMI CC 324262) was purified, and an N-terminal sequence and two internal sequences were determined. BglI genomic gene and the cDNA were cloned from a genomic library and by reverse transcriptase-polymerase chain reaction, respectively. The cDNA was successfully expressed in Saccharomyces cerevisiae and Pichia pastoris. Sequence analysis revealed that the gene encodes a 92-kDa enzyme that is a member of glycosidase family 3. 1 H-NMR analysis of the reaction catalyzed by this enzyme confirmed that, in common with other family 3 glycosidases, this enzyme hydrolyzes with net retention of anomeric configuration. Accordingly, the enzyme was inactivated by 2-deoxy-2-fluoro ␤-glucosyl fluoride, with kinetic parameters of k i ‫؍‬ 4. ␤-Glucosidases (EC 3.2.1.21; ␤-D-glucoside glucohydrolase) play a number of different important roles in biology, including the degradation of cellulosic biomass by fungi and bacteria, degradation of glycolipids in mammalian lysosomes, and the cleavage of glucosylated flavonoids in plants. These enzymes are therefore of considerable industrial interest, not only as constituents of cellulose-degrading systems, but also in the food industry (2, 3).Aspergillus species are known as a useful source of ␤-glucosidases (4 -6), and Aspergillus niger is by far the most efficient producer of ␤-glucosidase among the microorganisms investigated (4). Shoseyov et al. (7) have described a ␤-glucosidase from A. niger B1 (CMI CC 324626), which is active at low pH values as well as in the presence of high ethanol concentrations. This enzyme effectively hydrolyzes flavor compound glycosides in certain low pH products, such as wine and passion fruit juice, thereby enhancing their flavor (8 -11) and is particularly attractive for use in the food industry because A. niger is considered nontoxic (3). Other A. niger ␤-glucosidases have also been purified (12-14); however, differences in their properties have been reported, including ranges of molecular masses (116 -137 kDa) and isoelectric points (pI values of 3.8 -4) and pH optima (3.4 -4.5). Indeed, at least two ␤-glucosidases with distinct substrate specificities have been identified in commercial A. niger ␤-glucosidase preparations (15). To clear this confusion and also to allow protein engineering work to be performed it was important to clone, express, and characterize a ␤-glucosidase from this source. Although the cloning and expression of a functional A. niger ␤-glucosidase gene in Saccharomyces cerevisiae has been reported previously (16), the protein was not characterized, and the sequence was not published.Glycosidases have been assigned to families on the basis of sequence similarities, there now being some 77 different such families defined containing over 2000 different enzymes (17) With the exception of the glucosylceramidases (family 30) all simple ␤-glucosidases belong to either family 1 or 3. Family 1 contains enzymes from bacteria, plants, and mammals including also 6-phospho-glucosidases and thioglucosidas...

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confidence: 99%

Cloning, Expression, Characterization, and Nucleophile Identification of Family 3, Aspergillus nigerβ-Glucosidase

Dan¹,

Marton²,

Dekel³

et al. 2000

Journal of Biological Chemistry

125

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“…25) The pH stability (3 7) of the enzyme is within the range of the glucosidase of A. fumigatus ( 2 8) 12) and A. niger (2.5 9.0). 10) The optimum temperature (60 C) of the purified enzyme is similar to that of A. sojae (60 C) 13) but higher than that of A. niger (50 C).…”

Section: Discussionmentioning

confidence: 77%

“…The glucosidase from A. niger, 10) A. oryzae, 11) A. fumigatus 12) and A. sojae 13) have been purified and their characteristics recorded. Above all, black Aspergillus species are one of the best producers of food processing enzymes recognized as food additives, both from the standpoint of historical results and non pathogenicity.…”

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confidence: 99%

Purification, Characterization, and a Potential Application of .BETA.-Glucosidase from Aspergillus pulverulentus YM-80

間瀬¹,

森²,

横江³

2004

J. Appl. Glycosci.

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“…P-Glucosidase from Phanerochaete chrysosporium is more active on P-1 ~3-glucosidic linkages (26), whereas the purified enzyme from Thermitomyces clypeatus (27) displays moderate activity against gentiobiose, salicin and methyl-glucoside. The 13-glucosidases purified by Unno et al (28) and Watanabe et al (29) showed broader substrate specificity than the purified by us 13-gl ucos i dase.…”

Section: Substrate Specificity and Kinetic Analysismentioning

confidence: 80%

Purification and Biochemical Characteristics of β-Glucosidase from Aspergillus Awamori K-1

Petrova

Andreev

Bakalova

et al. 2002

Biotechnology & Biotechnological Equipment

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An extracellular f3-glucosidase (/3-D-glucoside glucohydrolase, EC 3.2.1.21) produced by Aspergillus awamori K-1 was purified to electrophoretic homogeneity. The purified enzyme, which had a glycoprotein nature, was a monomer with a relative molecular mass of 116 kDa and pi= 5.8. The f3-glucosidase was the most active against a1yl-f3-glucosides and cellobiose. In addition, the enzyme was able to catalyze the hydrolysis of fJ-1 ~2 and fJ-1 ~-linked diglucosides. The apparent Km and V values for p-nitrophenyl-/3-D-glucopyranoside were calculated to be 0.8 mM and 5.29 J.lmol.min-1 respectively. Maximal /3-glucosidase activity occurred at 60 oc and pH 5. 0. This enzyme was competitively inhibited by /3-D-glucose with Ki value of 13.15 mM

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High Recovery Purification and Some Properties of aβ-Glucosidase fromAspergillus niger

Cited by 34 publications

References 1 publication

Cloning, Expression, Characterization, and Nucleophile Identification of Family 3, Aspergillus nigerβ-Glucosidase

Cloning, Expression, Characterization, and Nucleophile Identification of Family 3, Aspergillus nigerβ-Glucosidase

Purification, Characterization, and a Potential Application of .BETA.-Glucosidase from Aspergillus pulverulentus YM-80

Purification and Biochemical Characteristics of β-Glucosidase from Aspergillus Awamori K-1

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