Jong Wan Yng Linda, Show Pau Loke, Ling Tau Chuan, Tan Yee Shin
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
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.
J Biosci Bioeng. 2017 Jul;124(1):91-98. doi: 10.1016/j.jbiosc.2017.02.008. Epub 2017 Mar 17.
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 (CuSO). 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 (KHPO) were tested. PEG 600 produced the highest top phase purification factor (P) of 1.33±0.62 with yield of 72.18±8.50%. The optimization of the ATPS parameters, such as volume ratio V, pH and crude enzyme loading are the factors controlling the phase partition. Our results showed that significant improvement (P of 6.26±2.87 with yield of 87.31±3.14%) of LiP recovery can be achieved by optimized the parameters.
皱盖乌芝是一种广泛分布于热带地区的野生蘑菇物种，属于担子菌纲。担子菌以其产生木质纤维素分解酶的能力而闻名，如木质素过氧化物酶（LiP）、漆酶（Lac）和锰过氧化物酶（MnP）。营养源、培养时间和搅拌等不同因素会影响木质纤维素分解酶的产生。皱盖乌芝在添加了马铃薯葡萄糖肉汤（PDB）、0.5%酵母和1.0%锯末的培养基中产生的LiP为26.70±3.31 U/mL。然而，当添加150μm的铜（硫酸铜）时，LiP活性增加到106.32±5.32 U/mL。双水相系统（ATPS）是一种用于LiP初步提取和回收的简单、快速且低成本的方法。共测试了由五种不同分子量的聚乙二醇（PEG）/磷酸氢二钾（KHPO）制成的25种系统。PEG 600产生的最高上相纯化因子（P）为1.33±0.62，产率为72.18±8.50%。双水相系统参数的优化，如体积比V、pH值和粗酶负载量是控制相分配的因素。我们的结果表明，通过优化参数可以显著提高LiP的回收率（P为6.26±2.87，产率为87.31±3.14%）。
