In vitro digestibility study of some plant protein sources as aquafeed for carps Labeo rohita and Cyprinus carpio using pH-Stat method.

Aquaculture, as a promising food industry, is expected to meet the demand for quality food from the increasing human population. As the diet is critical for feeding farm fish, such a faster growth in the industry is destined to create stress in the fishmeal market to supply diets to the tune. In this context, here, we studied the protein content of 20 plant ingredients, including aquatic weeds, cereals, pulses and oil-cakes using micro-Kjeldahl method and evaluated in vitro digestibility of these ingredients for rohu Labeo rohita and common carp Cyprinus carpio using pH-Stat method. The protein contents of water fern, duckweed, almond oil-cake and soybean product were 20.81, 39.75, 47.78 and 57.48%, respectively. Species-specific digestibility was found for the same plant ingredient. The degree of hydrolysis for water fern, duck weed, almond oil-cake and soybean product were 14.17, 4.80, 17.30 and 3.57%, respectively for rohu and 4.58, 6.03, 12.17 and 3.35%, respectively for common carp. This study showed that incorporation of water fern and almond oil-cake in the diet of rohu, and duck weed and almond oil-cake in the diet of common carp are beneficial considering their protein content and digestibility. These are cost-effective, protein-rich feed ingredients for aquafeed.

Trypsin from the digestive system of carp Cirrhinus mrigala: purification, characterization and its potential application.

Trypsin was purified 35.64-fold with 4.97% recovery from the viscera of carp Cirrhinus mrigala (mrigal) by ammonium sulfate precipitation, ion exchange and affinity chromatography. The purified enzyme was active at a wide range of pH (7.0-9.2) and temperature (10-50°C). The purified enzyme exhibited high thermal stability up to 50°C for 1h. The enzyme activity was stabilized by Ca(+2) (2mM) up to 7h at 40°C. The Km and kcat values of purified enzyme were 0.0672 mM and 92.09/s/mM, respectively. Soybean trypsin inhibitor and phenylmethylsulphonylflouride completely inhibited the enzyme activity. The specific inhibitor of trypsin, N-α-p-tosyl-L-lysine chloromethyl ketone inhibited 99.67% activity. Na(+), K(+) and Li(+) inhibited 20.99 ± 5.25%, 16.53 ± 4.80% and 18.99 ± 1.42% of enzyme activity, respectively. Divalent ions Mg(+2), Zn(+2), Co(+2), Hg(+2) and Cd(+2) inhibited 21.61 ± 2.22%, 31.62 ± 1.78%, 31.62 ± 1.96%, 85.68 ± 1.51% and 47.95 ± 2.13% enzyme activity, respectively. SDS-PAGE showed that the molecular mass of purified enzyme was 21.7 kDa. MALDI-TOF study showed a peptide sequence of AFCGGSLVNENKMHSAGHCYKSRIQV at the N-Terminal. This sequence recorded 76-84% identity with trypsin from Thunnus thynnus and other fish species. This confirmed that the purified protein was trypsin. The purified enzyme has potential applications in detergent and food industry because of its thermal stability and alkaline nature.