Journal of Environmental Biology

Size exclusion chromatography for the removal of pigments

from extracellular ligninolytic enzyme extracts

from decayed wheat straw

Author Details

Dharmendra Shukla

Department of Biosciences, Saurashtra University, Rajkot - 360 005, India

Bhavesh Patel

Department of Biosciences, Saurashtra University, Rajkot - 360 005, India

Hasmukh Modi

Department of Life Sciences, Gujarat University, Ahmedabad - 380 009, India

Bharat Rajiv Manuel Vyas

(Corresponding author)

Department of Biosciences, Saurashtra University, Rajkot - 360 005, India

e-mail: brmvyas@hotmail.com

Publication Data

Paper received:

12 April 2010

Revised received:

14 September 2010

Accepted:

24 September 2010

Abstract

Solid-state fermentation of wheat straw was carried out by a native white rot basidiomycete Daedaleopsis flavida strain 5A. Extract prepared from the 12-day decayed wheat straw contained extracellular ligninolytic enzymes like manganese peroxidase (MnP), manganese-independent peroxidase (MIP), lignin peroxidase (LiP) and laccase along with straw-degraded products and pigments. Sephacryl S-200 size exclusion chromatography in 16/100 column was used for the separation of these ligninolytic enzymes and straw-degraded products and pigments. Recovery of pigment-free ligninolytic enzyme activities as protein was 40% of the total proteins loaded and specific LiP activity increased 34 fold after size exclusion chromatography. Thus accurate estimation of LiP by veratryl alcohol oxidation assay was possible only after the removal of interfering pigments. The reproducibility of size exclusion chromatography is adjudged satisfactory from the consistent results obtained after seven repetitive uses of matrices.

Key words

Daedaleopsis flavida, Size exclusion chromatography, Extracellular ligninolytic enzymes, Lignin peroxidase, Pigments, Solid-state fermentation

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