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J Biol Chem

Title:		"Glucose oxidase from Aspergillus niger. Cloning, gene sequence, secretion from Saccharomyces cerevisiae and kinetic analysis of a yeast-derived enzyme"
Author(s):		Frederick KR; Tung J; Emerick RS; Masiarz FR; Chamberlain SH; Vasavada A; Rosenberg S; Chakraborty S; Schopfer LM;
Address:		"Chiron Research Labs, Chiron Corporation, Emeryville, California 94608"
Journal Title:		J Biol Chem
Year:		1990
Volume:		265
Issue:		7
Page Number:		3793 - 3802
DOI:
ISSN/ISBN:		0021-9258 (Print) 0021-9258 (Linking)
Abstract:		"The gene for Aspergillus niger glucose oxidase (EC 1.1.3.4) has been cloned from both cDNA and genomic libraries using oligonucleotide probes derived from the amino acid sequences of peptide fragments of the enzyme. The mature enzyme consists of 583 amino acids and is preceded by a 22-amino acid presequence. No intervening sequences are found within the coding region. The enzyme contains 3 cysteine residues and 8 potential sites for N-linked glycosylation. The protein shows 26% identity with alcohol oxidase of Hansenuela polymorpha, and the N terminus has a sequence homologous with the AMP-binding region of other flavoenzymes such as p-hydroxybenzoate hydroxylase and glutathione reductase. Recombinant yeast expression plasmids have been constructed containing a hybrid yeast alcohol dehydrogenase II-glyceraldehyde-3-phosphate dehydrogenase promoter, either the yeast alpha-factor pheromone leader or the glucose oxidase presequence, and the mature glucose oxidase coding sequence. When transformed into yeast, these plasmids direct the synthesis and secretion of between 75 and 400 micrograms/ml of active glucose oxidase. Analysis of the yeast-derived enzymes shows that they are of comparable specific activity and have more extensive N-linked glycosylation than the A. niger protein"
Keywords:		"Amino Acid Sequence Aspergillus niger/enzymology/*genetics Base Sequence *Cloning, Molecular *Genes, Fungal Glucose Oxidase/biosynthesis/*genetics/metabolism Kinetics Molecular Sequence Data Peptide Fragments/isolation & purification Plasmids Recombinant;"
Notes:		"MedlineFrederick, K R Tung, J Emerick, R S Masiarz, F R Chamberlain, S H Vasavada, A Rosenberg, S Chakraborty, S Schopfer, L M eng GM11106/GM/NIGMS NIH HHS/ Comparative Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. 1990/03/05 J Biol Chem. 1990 Mar 5; 265(7):3793-802"