Purification and Analytical Application of Vigna mungo Chitinase for Determination of Total Fungal Load of Stored Cereals

Abstract

A novel chitinase from urd bean (Vigna mungo) seeds was purified up to homogeneity and optimized with respect to its optimum working conditions of pH, temperature, and substrate concentration. Overall, 145-fold purification with 70% yield of the purified chitinase was achieved. The notable features of the purified enzyme were its appreciative substrate affinity as well as catalytic efficiency, high thermo stability (70% retention of initial activity at 70 °C after 60 min of continuous exposure), and pretty good storage stability (half-life of 45 days at 5 °C). The enzyme was used for determination of total chitin contents of the stored cereals that in the absence of any insect infestation, were considered to be directly proportional to the total fungal load of the tested samples. The method was linear up to 7.0 mM with 0.04 mM as the limit of detection. Percent recoveries of added chitin were < 90.0% and within-day and between-day coefficients of variations were > 3.0% for all the samples. Chitin values in stored cereal samples obtained by the present method and the popular DNS method showed good correlation, the value for coefficient of determination (R) being < 0.98. Overall, the method yielded acceptable sensitivity, reproducibility, and accuracy. Graphical Abstract ᅟ.

Nanoparticles of Lignins and Saccharides from Fishery Wastes as Sustainable UV-Shielding, Antioxidant, and Antimicrobial Biofillers

“…CH/OLNPs showed melting point values compatible with that of common bioplastics, such as poly­(lactic acid) (PLA: 120–175 °C), poly­(butylene succinate) (PBS: 107 °C), poly­(butylene succinate adipate) (PBSA: 83,7 °C), and poly­(butylene adipate terephthalate) (PBAT: 120 °C), making them a useful biofiller for advanced blends . The relative amount of CH and lignin embedded into nanoparticles was evaluated by the 3,5-dinitrosalicylic acid (DNSA) colorimetric assay. , The type of lignin and the molecular weight and DD of the saccharide affected the amount of the two components (Table ). CH aggregated with OL better than KL (Table , entries 10–16 versus entries 3–9), and the amount of embedded saccharide increased by increasing the value of DD (Table , entries 3, 6, and entries 10, 13 versus entries 8, 9 and entries 15, 16).…”

Domiciliation of Trichoderma asperellum Suppresses Globiosporangium ultimum and Promotes Pea Growth, Ultrastructure, and Metabolic Features

“…Chitin azure was used to measure chitinase at A 575 nm. The U of chitinase was expressed as the amount that increases the absorbance by 0.01 [ 29 ].…”

A chitinase with antifungal activity from naked oat (Avena chinensis) seeds

“…Using 1% (w/v) colloidal chitin as substrate, chitinase activity was determined by DNS method with measuring the reducing end group N‐acetylglucosamine (GlcNAc) (Preety & Hooda, ). Colloidal chitin was prepared by the method of Sandhya (Sandhya et al, ).…”