Glucose oxidase from Penicillium amagasakiense

Abstract

The complete amino acid sequence of glucose oxidase from Penicillium amagasakiense was determined by Edman degradation and mass spectrometry of peptide fragments derived from three different specific proteolytic digests and a cyanogen bromide cleavage. The complete sequence of each monomer comprises 587 amino acid residues, contains three cysteine residues, and seven potential N-glycosylation sites, of which at least five were confirmed to be glycosylated. Glucose oxidase from P. amagasakiense shows a high degree of identity (66%) and 79% similarity to glucose oxidase from Aspergillus niger, and is a member of the glucose-methanol-choline (GMC) oxidoreductase family. The tertiary structures of glucose oxidase from A. niger and cholesterol oxidase from Brevibacterium sterolicum were superimposed to provide a template for the sequence comparison of members of the GMC family. Keywords : glucose oxidase; Penicillium amagasakiense ; amino acid sequence; glucose-methanol-choline oxidoreductase ; consensus pattern.The glycoprotein glucose oxidase catalyses the oxidation of stable but has a sixfold higher affinity for β-D-glucose and a β-D-glucose to D-glucono-1,5-lactone with the concomitant re-tenfold higher specificity constant than glucose oxidase from duction of molecular oxygen to hydrogen peroxide. The enzyme Aspergillus niger (Kalisz et al., 1997). Despite the commercial has been purified from Aspergillus (Pazur and Kleppe, 1964; importance and extensive industrial application of glucose oxiSwoboda and Massey, 1965) and Penicillium species (Kusai et dase (Röhr et al., 1983;Crueger and Crueger, 1984; Wilson and al., 1960). Fungal glucose oxidase is a homodimer of approxi-Turner, 1992), and the availability of the preliminary X-ray difmately 160 kDa containing two tightly, but non-covalently, fraction data , the three-dimensional strucbound FAD cofactors (Yoshimura and Isemura, 1971a,b; Hay-ture of P. amagasakiense glucose oxidase has only recently been ashi and Nakamura, 1976) and 13% carbohydrate of the high elucidated (Wohlfahrt, G., Witt, S., Herdle, J., Kalisz, H. M., mannose type (Hayashi and Nakamura, 1976; Kalisz et al., Schomburg, D. and Hecht, H. J., unpublished results). The main 1997). Glucose oxidase from Penicillium amagasakiense is less stumbling block was the unavailability of its primary structure.In contrast, both the amino acid sequence (Kriechbaum et al., Correspondence to H. M. Kalisz, Gesellschaft für Biotechnologische 1989; Frederick et al., 1990) and X-ray structure (Hecht et al.,

Genes involved in the methyl tert-butyl ether (MTBE) metabolic pathway of Mycobacterium austroafricanum IFP 2012

“…3(b). We were able to detect the four highly conserved representative regions of the FAD-binding domain of choline dehydrogenases (Kiess et al, 1998). This FAD-binding domain contains the Walker-type-A motif previously detected and a characteristic b-sheet domain which was revealed in MpdB by the secondary structure prediction.…”

“…(b) The deduced amino acid sequence of MpdB was structurally aligned with the sequences of choline dehydrogenase from E. coli (Eco; Lamark et al, 1991), Sinorhizobium meliloti (Sme; Pocard et al, 1997) and the COG2303 consensus sequence, which includes choline dehydrogenases (betA genes) and related flavoproteins. Motifs were detected by similarity with consensus sequences of the GMC oxidoreductase family, described by Kiess et al (1998), and secondary structure (TOPO) predicted as described above. X X X X (Fig.…”

A survey of genes encoding H2O2-producing GMC oxidoreductases in 10 Polyporales genomes

“…2 AAO, GOX, MOX, CDH, respectively) with the exception of P2Os. This asparagine residue, also conserved in other GMCs, is involved in flavin bent conformation (Kiess et al 1998).…”

“…Two additional GMC enzymes, which are inefficient reducing O 2 to H 2 O 2 , are cellobiose dehydrogenase (CDH, EC 1.1.99.18) and pyranose dehydrogenase (PDH, EC 1.1.99.29) (Zámocký et al 2006, Kruså et al 2008, Peterbauer and Volc 2010. All members of the GMC superfamily share similar structural features (Wierenga et al 1986, Kiess et al 1998. Recently several GMCs have been classified in the so-called subfamilies AA3_1 (CDH), AA3_2 (AAO/GOX), AA3_3 (MOX) and AA3_4 (P2O) of the CAZy database (Levasseur et al 2013), but this nomenclature is not used here.…”

Cloning and Expression of Three Formate Oxidase Genes fromDebaryomyces vanrijiaeMH201

“…The two sequences were consistent with xhxhGxGxxGxxxhxxh(x) 6{9 hxhE (x is any residue and h is a hydrophobic residue) shown to be a conserved motif of the FAD binding site. 10) Also, VVDPALKVHGVKN-LRVIDASIIP and VVDPALKVHGVKHLRVIDASIIP, which were identical with that of one of two GMC oxidoreductase signature patterns, V(A)XDXXXXVX-GXXG(N)LR-(K/Y)VXDXSXXP, 11) were found among residues 523-545 of FOD1 and FOD2 and 3 respectively. FOD1 and FOD2 and 3 had VIVSQGVFESPK-LLMLSGIG and VILSQGVFESPKLLMLSGVG among residues of 259-278 respectively, and the sequences were identical with that of the other GMC signature patterns, VIL(V)XAGXXXSPXXLXXSGI(V)G, except that the fifth residue was different.…”