Purification and Characterization of Chitinase from Paenibacillus sp. D1

Abstract

A 56.56-kDa extracellular chitinase from Paenibacillus sp. D1 was purified to 52.3-fold by ion exchange chromatography using SP Sepharose. Maximum enzyme activity was recorded at pH 5.0 and 50 °C. MALDI-LC-MS/MS analysis identified the purified enzyme as chitinase with 60% similarity to chitinase Chi55 of Paenibacillus ehimensis. The activation energy (E (a)) for chitin hydrolysis and temperature quotient (Q (10)) at optimum temperature was found to be 19.14 kJ/mol and 1.25, respectively. Determination of kinetic constants k (m), V (max), k (cat), and k (cat)/k (m) and thermodynamic parameters ΔH*, ΔS*, ΔG*, ΔG*(E-S), and ΔG*(E-T) revealed high affinity of the enzyme for chitin. The enzyme exhibited higher stability in presence of commonly used protectant fungicides Captan, Carbendazim, and Mancozeb compared to control as reflected from the t (1/2) values suggesting its applicability in integrated pest management for control of soil-borne fungal phytopathogens. The order of stability of chitinase in presence of fungicides at 80 °C as revealed from t (1/2) values and thermodynamic parameters E (a(d)) (activation energy for irreversible deactivation), ΔH*, ΔG*, and ΔS* was: Captan > Carbendazim > Mancozeb > control. The present study is the first report on thermodynamic and kinetic characterization of chitinase from Paenibacillus sp. D1.

The Genomes, Proteomes, and Structures of Three Novel Phages That Infect the Bacillus cereus Group and Carry Putative Virulence Factors

“…The fourth protein (gp52) may aid Basilisk by allowing access to the cell, since it has significant similarity (41% identity over 251 amino acids) to a chitinase found in Paenibacillus sp. (65). A putative major capsid protein (MCP) was not identified by sequence comparison.…”

Cloning, expression, purification and application of a novel chitinase from a thermophilic marine bacterium Paenibacillus barengoltzii

“…They have gained a great deal of attention in recent years due to their diverse properties and broad range of potential industrial applications (Bhattacharya, Nagpure, & Gupta, 2007). For example, they can be used to produce N-acetyl COSs and GlcNAc in food industry (Singh & Chhatpar, 2011), convert chitin wastes to bio-ethanol in energy industry (Purushotham & Podile, 2012), and biologically control fungal pathogens in the field of agriculture (Patil & Jadhav, 2015).…”

Production and Characterization of Chitinases from Thermophilic Bacteria Isolated from Prataan Hot Spring, East Java

“…98CJ11027 [33] and Paenibacillus sp. D1 [34], which were mostly active at acidic pH. The optimum pH determined in this experiment was used for further experiments.…”

“…D1 and Bacillus sp. HSA,3-1a, which had optimum activity at 50 °C and 60 °C respectively [34,25]. Table 2 summarizes the results of each purification step of chitinase from Paenibacillus sp.…”

“…Two protein fractions were obtained from SDS-PAGE with molecular weights of 68 and 82 kDa. These results are similar to previously reported data from another Paenibacillus sp., where the chitinases had molecular masses ranging from 40 to 150 kDa [34,36].…”