Journal of Agricultural Engineering and Food Technology Print ISSN: 2350-0085; Online ISSN: 2350-0263; Volume 2, Number 2; April-June, 2015 pp. 131-135 © Krishi Sanskriti Publications http://www.krishisanskriti.org/jaeft.html Heat Tolerant Superoxide Dismutase from Therapeutically Significant Cuminum cyminum: Partial Purification and Kinetic Studies Shikha Agarwal 1* , Priyanka Sharma 2 and Nidhee Chaudhary 3 1,2,3 Amity Institute of Biotechnology, Amity University Uttar Pradesh Sector- 125, Noida- 201313, India E-mail: 1* shikhu1904@gmail.com, 2 priyankasharma.bscmicro@gmail.com, 3 nchaudhary@amity.edu Abstract—Superoxide dismutase (SOD, EC 1.15.1.1), an antioxidant enzyme; strengthens body’s primary antioxidant system playing a key role in scavenging the free radicals (disease causing agents) and helps fighting the diseases such rheumatoid arthritis, neurological disorders, Alzheimer’s disease, Diabetes, Parkinson’s disease, heart disease and cancer. Since the digestive tract enzymes present in human body easily deactivate SOD molecules, great need develops for bioavailable forms of SOD. In the present investigation, Cuminum cyminum (Zeera) has been studied for the presence of high specific activity of SOD. C. cyminum or Cumin seeds, used in Indian food on almost daily basis, are well known for great health benefits such as enhancing appetite, digestion, vision, strength and lactation. The specific activity of the crude enzyme extracted from C. cyminum was found to be 0.57 Units/mg. The SOD enzyme obtained has been partially purified into three fractions based on 0-30%, 30-60% and 60-90% saturation level of (NH 4 ) 2 SO 4 ; 0-30% fraction (5.46 Units/mg) having higher specific activitywith purification fold of 9.57. The screened enzyme fraction after dialysis was further characterized on the basis of various biochemical parameters. The enzyme was found to show maximum activity at pH 8.0, temperature 30 0 C in 30 minutes of reaction time. The thermal and pH stability values of the enzyme were 50 0 C and 6.0-9.0, respectively. The K m and V max values were determined to be 0.284 mM and 2.142 inhibition percent ml -1 min -1 , respectively. Various ions in the form of their respective salts were used to perform inhibition studies. They were found to inhibit SOD activity in the given descending order; Na + >Mg 2+ >Ca 2+ >Zn 2+ . The results obtained suggest that C. cyminum, a medicinal important plant, can serve as potential source of heat tolerant SOD, which further have great importance in therapeutics and other bio-industries. Keywords: Cuminum cyminum, Superoxide dismutase, Heat tolerant, Antioxidant, Therapeutics. 1. INTRODUCTION Reactive oxygen species (ROS) are small, highly reactive, oxygen-containing molecules (free radicals) that are naturally generated in small amounts during the body’s metabolic reactions [1]. These free radicals can react and damage complex cellular molecules such as fats, proteins, or DNA. The cellular damage caused by free radicals, initiates accumulation of mutations in nucleic acids sequences, which increases in number by time and finally gives rise to rapid aging, rheumatoid arthritis, heart disease, Parkinson’s disease, Diabetes, Alzheimer’s disease, neurological disorders and cancer [2]. Because ROS form naturally during many metabolic processes, cells have developed several protective mechanisms to detoxify the ROS. Co-operative defense systems that protect the body from free radical damage include the anti-oxidant nutrients and enzymes [3]. Antioxidant plays an important role in human health because the biological defense mechanism cannot operate under severe oxygen stress. Nature has endowed us with protective antioxidant mechanisms- superoxide dismutase (SOD), catalase, glutathione peroxidases etc, which protects us from metabolic damages [4]. Superoxide Dismutase (SOD: 1.15.1.1) an antioxidant enzyme omnipresent in all living cell inner and outer membranes acts as catalyst in dismutation of Superoxide (O 2 -) radical into hydrogen peroxide and molecular oxygen and helps in scavenging these free radicals to fight against the diseases [5]. Since the digestive tract enzymes present in human body easily deactivate SOD molecules, a high need is developed of finding bioavailable forms of SOD [6]. High Intake of fruits and vegetable can promote longevity and offers a number of health benefits against degenerative diseases. Intake of medicinal plants in rats results in an increase in of activity superoxide dismutase [7]. Plant producing significant amount of SOD enzyme play crucial role against various pathological diseases. Various sources have been found to contain substantial amount of SOD viz; spices [8], dry fruits [9] and flowering plants [10]. Cuminum Cyminum, belongs to family Apiaceae and is extensively used in culinary practices of the Indian Subcontinent and some other Asian, African and Latin American countries as a spice [11].Cumin (zeera) seeds have been used as antipyretic,