Characterization of lignin-degrading enzymes (LDEs) from a dimorphic novel fungus and identification of products of enzymatic breakdown of lignin.
Lignin is a major component of all plants, the degradation of which remains a major challenge to date owing to its recalcitrant nature. Several classes of fungi have been studied to carry out this process to some extent, but overall the process remains inefficient. We have isolated a novel alkalophilic dimorphic lignin-degrading Deuteromycete from soil, identified as "uncultured" and coded as MVI.2011. Supernatant from 12-h culture of MVI.2011 in optimized mineral medium containing lignin pH 9.0 was analysed for Lignin Peroxidase, Manganese Peroxidase and Laccase. Enzyme purification was carried out by standard protocols using ammonium sulphate precipitation followed by further purification by Gel Permeation Chromatography. Analysis of total protein, specific enzyme activity and molecular weight of the GPC-purified LiP, MnP and Laccase showed 93.83 μg/ml, 5.27 U/mg, 42 kDa; 78.13 μg/ml, 13.18 U/mg, 45 kDa and 85.81 μg/ml, 4.77 U/mg, 62 kDa, respectively. The purified enzymes possessed high activity over a wide range of pH (4-11), and temperature (30-55 °C). The optimum substrate concentration was 20 μg/ml of lignin for all the three enzymes. CD spectra suggested that the predominant secondary structure was helix in LiP, and, turns in MnP and Laccase. The breakdown products of lignin degradation by MVI.2011 and the three purified enzymes were detected and identified by FTIR and GC-MS. They were oxalic acid, hentriacontane, derivatives of octadecane, nonane, etc. These vital compounds are certain to find application as biofuels, an alternate energy source in various industries.
School of Bio Sciences and Technology, VIT University, Vellore, 632014, Tamil Nadu, India.|Department of Biotechnology, Prathyusha Institute of Technology and Management, Tiruvallur, 602025, Tamil Nadu, India.
Lipin Dev Mundur
LDM
Sahadevan
School of Bio Sciences and Technology, VIT University, Vellore, 632014, Tamil Nadu, India.|, Avenida Rovisco Pais, 1, 1049-001, Lisbon, Portugal.
Chandra Shekhar
CS
Misra
School of Bio Sciences and Technology, VIT University, Vellore, 632014, Tamil Nadu, India. dr.thankamani@gmail.com.
V
V
Thankamani
Biofuels; Laccase; Lignin; Lignin peroxidase; MVI.2011; Manganese peroxidase