+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Mutations of Thr169 affect substrate specificity of pyranose 2-oxidase from Trametes multicolor



Mutations of Thr169 affect substrate specificity of pyranose 2-oxidase from Trametes multicolor



Biocatalysis and Biotransformation 26(1-2): 120-127



Site-directed mutagenesis was used to enhance the catalytic activity of pyranose 2-oxidase (P2Ox) from Trametes multicolor with different substrates. To this end, threonine at position 169 was replaced by glycine, alanine and serine, respectively. Using oxygen as electron acceptor the mutant T169G was equally active with d-glucose and d-galactose, whereas wild-type recombinant P2Ox only showed 5.2% relative activity with the latter substrate. When d-galactose was used as electron donor in saturating concentrations, T169G showed a 4.5-fold increase in its catalytic efficiency k(cat)/K-M for the alternative electron acceptor 1,4-benzoquinone and a nine-fold increased k(cat)/K-M value with the ferricenium ion compared with wt recP2Ox. Variant T169S showed an increase in its catalytic efficiency both with 1,4-benzoquinone (3.7 times) as well as with the ferricenium ion (1.4 times) when D-glucose was the substrate.

(PDF emailed within 0-6 h: $19.90)

Accession: 021385320

Download citation: RISBibTeXText

DOI: 10.1080/10242420701789320


Related references

Oxidation of Phe454 in the Gating Segment Inactivates Trametes multicolor Pyranose Oxidase during Substrate Turnover. Plos One 11(2): E0148108, 2016

Production of a novel pyranose 2-oxidase by basidiomycete Trametes multicolor. Applied Biochemistry and Biotechnology 70-72: 237-248, 1998

Kinetic mechanism of pyranose 2-oxidase from trametes multicolor. Biochemistry 48(19): 4170-4180, 2009

Biochemical characteristics of Trametes multicolor pyranose oxidase and Aspergillus niger glucose oxidase and implications for their functionality in wheat flour dough. Food Chemistry 131(4): 1485-1492, 2012

Purification and characterization of pyranose oxidase from the white rot fungus Trametes multicolor. Applied and Environmental Microbiology 67(8): 3636-3644, 2001

The bread dough stability improving effect of pyranose oxidase from trametes multicolor and glucose oxidase from Aspergillus niger: unraveling the molecular mechanism. Journal of Agricultural and Food Chemistry 61(32): 7848-7854, 2014

Impact of pyranose oxidase from Trametes multicolor, glucose oxidase from Aspergillus niger and hydrogen peroxide on protein agglomeration in wheat flour gluten-starch separation. Food Chemistry 148: 235-239, 2014

Crystallization and preliminary X-ray diffraction analysis of pyranose 2-oxidase from the white-rot fungus Trametes multicolor. Acta Crystallographica. Section D, Biological Crystallography 60(Pt 1): 197-199, 2003

Conformational heterogeneity in pyranose 2-oxidase from Trametes multicolor revealed by ultrafast fluorescence dynamics. Journal of Photochemistry and Photobiology A: Chemistry 234: 44-48, 2012

Identification of the covalent flavin adenine dinucleotide-binding region in pyranose 2-oxidase from Trametes multicolor. Analytical Biochemistry 314(2): 235-242, 2003

A thermostable triple mutant of pyranose 2-oxidase from Trametes multicolor with improved properties for biotechnological applications. Biotechnology Journal 4(4): 525-534, 2009

Probing active-site residues of pyranose 2-oxidase from Trametes multicolor by semi-rational protein design. Biotechnology Journal 4(4): 535-543, 2009

Enzymatic formation of dicarbonyl sugars: C-2 oxidation of 1->6 disaccharides gentiobiose, isomaltose and melibiose by pyranose 2-oxidase from Trametes multicolor. Journal of Carbohydrate Chemistry 18(8): 999-1007, 1999

Enzymatic formation of dicarbonyl sugars: C-2 oxidation of 1 leads to 6 disaccharides gentiobiose, isomaltose and melibiose by pyranose 2-oxidase from Trametes multicolor. Journal of carbohydrate chemistry8(8): 999-1007, 1999

Evaluation of different expression systems for the heterologous expression of pyranose 2-oxidase from Trametes multicolor in E. coli. Microbial Cell Factories 9: 14, 2010