Isolation, purification and characterization of acid phosphatase from Chara sp.

摘要

Algae are also very important ecologically because, used in production of many economical valuable products and they are beginning of the food chain for other animals. Whereas, ample of reports are available in morphological studies on fresh water algae throughout the globe. However, enzyme research of algal origin is ephemeral. Hydrolytic enzymes have great reasonable significance owing to their central role in biological processes. In this study, acid phosphatase enzyme characterized from Chara sp. fresh water alga isolated province Waghur River located near Skegaon, district Jalgaon, Maharashtra (India). The acid phosphatase is a monomer protein purified by ion-exchange and gel filtration to 3.87 fold with an apparent molecular mass 66 kDa on SDS PAGE. The purified acid phosphatase has an optimum pH of 4.0, and optimum temperature for the hydrolysis of p-Nitro phenyl phosphate at 50oC and the km and Vmax 0.25 mm and 0.012 mu;mol minndash;1mgndash;1 respectively at the same conditions. The activation energy found to be 54.20 KJ/mole and Q10 value was 2.08 between 40 and 50deg;C and fairly stable at temperature up to 37deg;C. The activity of the enzyme enhanced by EDTA, Tween 20, Triton-X and Guaiacol and heavy metals Fe3+, Cu2+, Ca2+, Mg2+, Hg2+ and K+. The enzyme strongly inhibited by organic solvents, SDS, tween 20 and heavy metals Zn2+. The present article reveals on bio-molecular characterization of acid phosphatase with kinetic studies.