Recovery of lignin peroxidase from submerged liquid fermentation of Amauroderma rugosum (Blume & T. Nees) Torrend using polyethylene glycol/salt aqueous two-phase system.
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 106.32±5.32 U/mL when supplemented with 150 μm 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 (PFT) of 1.33±0.62 with yield of 72.18±8.50%. The optimization of the ATPS parameters, such as volume ratio VR, pH and crude enzyme loading are the factors controlling the phase partition. Our results showed that significant improvement (PFTof 6.26±2.87 with yield of 87.31±3.14%) of LiP recovery can be achieved by optimized the parameters.
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Wan Yng Linda
WYL
Jong
Department of Chemical and Environmental Engineering, Faculty of Engineering, University of Nottingham Malaysia Campus, Jalan Broga, 43500 Semenyih, Selangor Darul Ehsan, Malaysia; Manufacturing and Industrial Processes Division, Faculty of Engineering, Centre for Food and Bioproduct Processing, University of Nottingham Malaysia Campus, Jalan Broga, 43500 Semenyih, Selangor Darul Ehsan, Malaysia.
Pau Loke
PL
Show
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Tau Chuan
TC
Ling
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia; Mushroom Research Centre, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: tanyeeshin@um.edu.my.
Yee Shin
YS
Tan
D005591:Chemical Fractionation / Q000379:methods*; D005285:Fermentation; D007101:Immersion; D008970:Molecular Weight; D010544:Peroxidases / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry*; D020072:Polyporales / Q000378:metabolism*; D012965:Sodium Chloride / Q000737:chemistry; D014867:Water / Q000737:chemistry*
D014867:Water; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D010544:Peroxidases; C042858:lignin peroxidase
Aqueous two-phase system; Lignin peroxidase; Mushroom; Polyethylene glycol; Purification factor; Submerged fermentation