Year-end Sale % OFF 
Abstract
The alcohol oxidase (AOx) cDNA from Aspergillus terreus MTCC6324 with an open reading frame (ORF) of 2001 bp was constructed from n-hexadecane induced cells and expressed in Escherichia coli with a yield of ∼4.2 mg protein g−1 wet cell. The deduced amino acid sequences of recombinant rAOx showed maximum structural homology with the chain B of aryl AOx from Pleurotus eryngii. A functionally active AOx was achieved by incubating the apo-AOx with flavin adenine dinucleotide (FAD) for ∼80 h at 16°C and pH 9.0. The isoelectric point and mass of the apo-AOx were found to be 6.5±0.1 and ∼74 kDa, respectively. Circular dichroism data of the rAOx confirmed its ordered structure. Docking studies with an ab-initio protein model demonstrated the presence of a conserved FAD binding domain with an active substrate binding site. The rAOx was specific for aryl alcohols and the order of its substrate preference was 4-methoxybenzyl alcohol >3-methoxybenzyl alcohol>3, 4-dimethoxybenzyl alcohol > benzyl alcohol. A significantly high aggregation to ∼1000 nm (diameter) and catalytic efficiency (kcat/Km) of 7829.5 min−1 mM−1 for 4-methoxybenzyl alcohol was also demonstrated for rAOx. The results infer the novelty of the AOx and its potential biocatalytic application.
Highlights
The alcohol oxidase (AOx) enzymes (Alcohol: O2 Oxidoreductase; EC 1.1.3.x) catalyze the oxidation of various alcohol substrates to the corresponding carbonyl compounds with a concomitant release of hydrogen peroxide
The results clearly demonstrates the substrate affinity for our predicted AOx model for aromatic alcohols in order of preference with its total binding energy in parenthesis as 4-methoxybenzyl alcohol (282.87 kJ mol21) .3methoxybenzyl alcohol (274.12 kJ mol21).3, 4 dimethoxybenzyl alcohol (271.75 kJ mol21). benzyl alcohol (256.45 kJ mol21)
The functional role of the AOxs from A.terreus is recognized as catalyst for the oxidation of the intermediate alcohol substrates formed during the cytochrome P450 (CYP450) catalyzed degradation of various hydrocarbon substrates [45]
Summary
The alcohol oxidase (AOx) enzymes (Alcohol: O2 Oxidoreductase; EC 1.1.3.x) catalyze the oxidation of various alcohol substrates to the corresponding carbonyl compounds with a concomitant release of hydrogen peroxide. Over the last decade these flavoenzymes have attracted wide attention for their potential industrial applications due to the fact that they can catalyze the oxidation of alcohol substrates irreversibly and selectively, and they do not require any external co-factors for the catalysis since the cofactor, which is flavin or their derivative, is avidly linked to the protein matrix of these redox enzymes. These groups of enzymes are available in nature because various aerobic microorganisms produce them and offer the scope for their large scale production using suitable bioreactors. The sources reported for these enzymes are mostly bacteria, yeast, fungi, plant, insect and mollusks
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
