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Titolo:
Purification of alpha-N-acetyl-galactosaminidase from Aspergillus niger and its use in the synthesis of GalNAc-alpha-(1 -> O)-serine
Autore:
Naundorf, A; Ajisaka, K;
Indirizzi:
Meiji Milk Prod Co Ltd, Inst Hlth Sci, Odawara, Kanagawa 2500862, Japan Meiji Milk Prod Co Ltd Odawara Kanagawa Japan 2500862 gawa 2500862, Japan
Titolo Testata:
ENZYME AND MICROBIAL TECHNOLOGY
fascicolo: 6, volume: 25, anno: 1999,
pagine: 483 - 488
SICI:
0141-0229(199909)25:6<483:POAFAN>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Keywords:
alpha-N-acetyl-galactosaminidase; Aspergillus niger; purification; O-glycopeptide conjugate; enzymatic synthesis; gram-scale;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
10
Recensione:
Indirizzi per estratti:
Indirizzo: Ajisaka, K Meiji Milk Prod Co Ltd, Inst Hlth Sci, 540 Naruda, Odawara, Kanagawa 2500862, Japan Meiji Milk Prod Co Ltd 540 Naruda Odawara Kanagawa Japan 2500862
Citazione:
A. Naundorf e K. Ajisaka, "Purification of alpha-N-acetyl-galactosaminidase from Aspergillus niger and its use in the synthesis of GalNAc-alpha-(1 -> O)-serine", ENZYME MICR, 25(6), 1999, pp. 483-488

Abstract

alpha-N-Acetyl-galactosaminidase was purified from several commercially available Aspergillus niger enzyme fractions. By using GalNAc-alpha-pNP as the substrate, a K-m = 0.23 mM and V-max = 210 mu mol/min mg protein and a pH-optimum in the range of 2.6-3.2 were determined. These enzyme preparationswere tested under conditions of reverse hydrolysis with unprotected L-serine as the acceptor. Incubation conditions in the preparative synthesis of GalNAc-alpha-(1 --> O)-serine were optimized with respect to incubation timeand pH. By using 80 U of enzyme/mmol of starting material at pH 3.5, thermodynamic equilibration of the incubation mixture was reached after 4 days. To scale up the synthesis to the gram-scale, a new purification procedure was developed based on column chromatography on activated carbon. Because the starting material N-acetyl-galactosamine is itself an expensive component, we also examined the recovery of unconverted galactosamine. Recovery rates were determined to be in the range of 60-70%. (C) 1999 Elsevier Science Inc. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/10/20 alle ore 04:09:53