Multisubstrate specific flavin containing monooxygenase from Chlorella pyrenoidosa with potential application for phenolic wastewater remediation and biosensor application.

Abstract:

:Microbial degradation of phenolic pollutants in industrial wastewater is dependent on enzymatic pathway comprising a cascade of phenol metabolizing enzymes. Phenol hydroxylase is the first enzyme of the pathway catalysing the initial attack on phenol in green algae Chlorella pyrenoidosa. The present work reports cost-effective production of partially purified microalgal phenol hydroylase by single-step purification and characterization of its kinetic properties with the view of application for enzyme-based remediation of phenolic wastewater or in phenolic biosensor. The enzyme with a molecular weight of 25 kDa shows all characteristics of phenol hydroxylases, that is, hydroxylation of phenol to catechol (confirmed by HPLC), substrate-dependent NADPH oxidation, absorption spectrum typical of flavoproteins and peptide mass fingerprint corresponding to flavoprotein hydroxylase. The enzyme utilizes phenol with apparent Michealis constant (Km) of 1.71 µM, maximal velocity (Vmax) of 0.4 µM/min with optimal activity at pH 7 and 35°C. Fe2+chelators (Phenanthroline and sodium arsenate), heavy metals, denaturants and oxidizing agents showed inhibitory effect on phenol hydroxylation activity of the enzyme. The enzyme has broad substrate specificity against isomeric diphenols, isomeric methylphenols, halogen-substituted phenols, amino-substituted phenols, nitrophenols, hydroxybenzaldehyde and hydroxylbenzoic acid. The enzyme shows remarkable storage stability at room temperature and at 4°C. The multisubstrate specificity coupled to remarkable storage stability of the microalgal phenol hydroxylase opens up avenues for its application in remediation of a wide range of phenolics released in industrial wastewater or phenolic biosensor application.

journal_name

Environ Technol

journal_title

Environmental technology

authors

Das B,Patra S

doi

10.1080/09593330.2017.1349838

subject

Has Abstract

pub_date

2018-08-01 00:00:00

pages

2073-2089

issue

16

eissn

0959-3330

issn

1479-487X

journal_volume

39

pub_type

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