Optimization and characterization of trypsin of Labeo rohita from its visceral waste

Charu Batav; Ragini Gothalwal

doi:10.30574/gscarr.2020.3.2.0041

Authors

Charu Batav Department of Biotechnology, Barkatullah University, Bhopal, M.P. India.
Ragini Gothalwal Department of Biotechnology, Barkatullah University, Bhopal, M.P. India.

DOI:

https://doi.org/10.30574/gscarr.2020.3.2.0041

Keywords:

Fish visceral waste, Protease, Trypsin, SDS-PAGE, LCMS, Enzyme kinetics

Abstract

The disposal of fish visceral waste, is currently posing a major threat to fishery industries and environment. 35% of the total global catch and 27% of the landed fish is wasted. The inappropriate disposal of fish waste pollutes ground water and soil (by nitrate leaching), surface water (by eutrophication) and harbors insect/rodent infestations due to release of noxious odor such as indole, sketol, cadavarine, putricine etc. Technical advancement has enabled recovery of value-added products. Present investigation involved isolation of visceral enzyme from widely consumed freshwater fish Labeo rohita. The enzyme was purified and characterized as trypsin. Among the different precipitating agents chosen cold acetone was found the best. The amidolytic activity of the enzyme using BAPNA substrate was studied. From the Lineweaver-Burk plot the Km and Vmax were calculated. The molecular weight, optimum pH and temperature were 24 KDa, 8.5 and 40ºC, resp. The LCMS studies showed that the enzyme exhibited 19.3% similarity with commercial (bovine) trypsin. It was thus, anticipated to minimize the waste burden in an eco-friendly manner.

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