4.4 Article

Recovery of lignin peroxidase from submerged liquid fermentation of Amauroderma rugosum (Blume & T. Nees) Torrend using polyethylene glycol/salt aqueous two-phase system

Journal

JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 124, Issue 1, Pages 91-98

Publisher

SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2017.02.008

Keywords

Aqueous two-phase system; Lignin peroxidase; Polyethylene glycol; Purification factor; Mushroom; Submerged fermentation

Funding

  1. Fundamental Research Grant Scheme (Malaysia) from the Ministry of Higher Education Malaysia [FRGS/1/2015/SG05/UNIM/03/1]
  2. Mushroom Research Centre grant [UM.0000117-KWIAK]

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Amauroderma rugosum is a wild mushroom species widely distributed in tropics and is classified under the class of Basidiomycetes. Basidiomycetes are well-known for their abilities of producing lignocellulolytic enzymes such as lignin peroxidase (LiP), laccase (Lac) and manganese peroxidase (MnP). Different factors such as nutrient sources, incubation period and agitation affect the production of lignocellulolytic enzymes. The A. rugosum produced LiP in the medium supplemented with potato dextrose broth (PDB), 0.5% yeast and 1.0% saw dust at 26.70 +/- 3.31 U/mL. However, the LiP activity was increased to 10632 +/- 5.32 U/mL when supplemented with 150 mu m of copper (CuSO4). The aqueous two-phase system (ATPS) is a simple, rapid and low cost method for primary extraction and recovery of LiP. A total of 25 systems made from five different molecular weights of polyethylene glycol (PEG)/dipotassium hydrogen phosphate (K2HPO4) were tested. PEG 600 produced the highest top phase purification factor (P-FT) of 133 +/- 0.62 with yield of 72.18 +/- 8.50%. The optimization of the ATPS parameters, such as volume ratio V-R, pH and crude enzyme loading are the factors controlling the phase partition. Our results showed that significant improvement (P-FT of 6.26 +/- 2.87 with yield of 87.31 +/- 3.14%) of LiP recovery can be achieved by optimized the parameters. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

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