A novel continuous enzyme coupled colorimetric assay for phospholipase A2 and its application in the determination of catalytic activity of oil body associated oleosin protein

Title : A novel continuous enzyme coupled colorimetric assay for phospholipase A2 and its application in the determination of catalytic activity of oil body associated oleosin protein

Abstract:

The present work reports a novel, continuous, specific and colorimetric coupled method for the assay of phospholipase A₂ (PLA₂), which involves use of acyl-CoA synthetase (ACS) as coupling enzyme and Dilinoleoyl Phosphatidylcholine (DLPC) as substrate. This method is based on the principle that free fatty acids (FFA) produced by PLA₂ are acted upon by ACS, which requires Coenzyme A (CoA) as co-substrate. The PLA₂ activity was measured in terms of amount of CoA utilized in the coupled reaction, determined indirectly by measuring unreacted CoA using 5,5’-dithiobis (2-nitrobenzoate). The PLA₂ assay was in agreement with Michaelis–Menten equation (Vm = 11.9 nmole/min/mg), Km = 5.3 nmole, Vm/Km = 2.2 min^-1mg^-1). Compared to other available methods, this method provides an exact measure of PLA₂ as natural PLs are used as substrate in place of PL analogues. The embedded advantages of this method would have wider acceptability and applicability.

Biography:

Dr. Sweta Kumari Holding the Position of Scientist at ICAR- Indian Agricultural Research Institute (IARI), New Delhi. She Graduated in Agricultural Science from Banaras Hindu University (BHU), Varanasi and Post Graduates (Master’s and Ph.D.) in Biochemistry from ICAR- IARI, New Delhi. She is a Life Member of Society of Journal of plant Biochemistry and Biotechnology, New Delhi and Society of Indian Journal of Plant Physiology, New Delhi. She received much recognition in terms of awards and fellowship. She awarded two times with ICAR- NET by Agricultural Scientist Recruitment Board (ASRB), New Delhi, two times with JRF-NET by Council of Scientific and Industrial Research (CSIR) New Delhi, IARI –SRF by ICAR- Indian Agricultural Research Institute (IARI), New Delhi, ICAR-JRF (Junior Research Fellowship) for M.Sc by Indian Council of Agricultural Research (ICAR), New Delhi and UG-NTS (Undergraduate National Talent Scholarship) for B.Sc (Agri) by Indian Council of Agricultural Research (ICAR), New Delhi. She published several articles in the area of enhancement of nutrition and flavor quality of crop plants as plant food for improved human nutrition and health in different national and international journals and magazines.