Membrane chromatography for the purification of laccase from the supernatant of Pleurotus sapidus

Mayuratheepan Puthirasigamany; Monika Wirges; Tim Zeiner

doi:10.1016/j.bej.2012.11.003

Membrane chromatography for the purification of laccase from the supernatant of Pleurotus sapidus

Mayuratheepan Puthirasigamany, Monika Wirges, Tim Zeiner^*

^*Corresponding author for this work

Institute of Chemical Engineering and Environmental Technology (6670)

Research output: Contribution to journal › Article › peer-review

Abstract

Extracellular laccase from the fermentation broth of Pleurotus sapidus was purified using membrane chromatography. Five laccase iso-enzymes produced in submerged cultures were purified; the enzymes had isoelectric points (p. I) ranging from 3.3 to 4.7 and a calculated molecular weight of 57.4. kDa. The fermentation broth had a low product titre of 80 μg/l. Hydrophobic interaction (HIC) and ion exchange (IEX) membranes chromatography modules were used in an ÄKTApurifier 100 liquid chromatography system for the dynamic experiments. The pre-treatment of the fermentation broth was not required. The enzyme activity was retained, and enrichment up to 15-fold was observed via activity assays and SDS-Page analysis.

Original language	English
Pages (from-to)	180-187
Number of pages	8
Journal	Biochemical Engineering Journal
Volume	70
DOIs	https://doi.org/10.1016/j.bej.2012.11.003
Publication status	Published - 15 Jan 2013

Keywords

Downstream processing
Laccase
Membrane adsorption
Membrane chromatography
Pleurotus sapidus

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biomedical Engineering
Environmental Engineering

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10.1016/j.bej.2012.11.003

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title = "Membrane chromatography for the purification of laccase from the supernatant of Pleurotus sapidus",

abstract = "Extracellular laccase from the fermentation broth of Pleurotus sapidus was purified using membrane chromatography. Five laccase iso-enzymes produced in submerged cultures were purified; the enzymes had isoelectric points (p. I) ranging from 3.3 to 4.7 and a calculated molecular weight of 57.4. kDa. The fermentation broth had a low product titre of 80 μg/l. Hydrophobic interaction (HIC) and ion exchange (IEX) membranes chromatography modules were used in an {\"A}KTApurifier 100 liquid chromatography system for the dynamic experiments. The pre-treatment of the fermentation broth was not required. The enzyme activity was retained, and enrichment up to 15-fold was observed via activity assays and SDS-Page analysis.",

keywords = "Downstream processing, Laccase, Membrane adsorption, Membrane chromatography, Pleurotus sapidus",

author = "Mayuratheepan Puthirasigamany and Monika Wirges and Tim Zeiner",

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doi = "10.1016/j.bej.2012.11.003",

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AU - Puthirasigamany, Mayuratheepan

AU - Wirges, Monika

AU - Zeiner, Tim

PY - 2013/1/15

Y1 - 2013/1/15

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AB - Extracellular laccase from the fermentation broth of Pleurotus sapidus was purified using membrane chromatography. Five laccase iso-enzymes produced in submerged cultures were purified; the enzymes had isoelectric points (p. I) ranging from 3.3 to 4.7 and a calculated molecular weight of 57.4. kDa. The fermentation broth had a low product titre of 80 μg/l. Hydrophobic interaction (HIC) and ion exchange (IEX) membranes chromatography modules were used in an ÄKTApurifier 100 liquid chromatography system for the dynamic experiments. The pre-treatment of the fermentation broth was not required. The enzyme activity was retained, and enrichment up to 15-fold was observed via activity assays and SDS-Page analysis.

KW - Downstream processing

KW - Laccase

KW - Membrane adsorption

KW - Membrane chromatography

KW - Pleurotus sapidus

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JO - Biochemical Engineering Journal

JF - Biochemical Engineering Journal

SN - 1369-703X

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Membrane chromatography for the purification of laccase from the supernatant of Pleurotus sapidus

Abstract

Keywords

ASJC Scopus subject areas

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