Isocitrate lyase has been extracted from castor endosperm and purified about 70 fold with an overall recovery of about 28%. The purified ICL shows a single protein band on gel electrophoresis and only one wide band on SDS-PAGE. T he enzyme is eluted as a single symmetrical peak through 2.5x 50 cm column of sephadex G-200-120 molecular weight of enzyme is estimated to be 140,000 from gel filtration. The sub unit molecular weight has been found to be 34,000 to 35,000. Thus the enzyme is tetrameric protein made up of apparently identical subunits. Thermal inactivation of purified enzyme at 400 & 450 shows biphasic kinetics in which half of the activity is destroyed quiet fast and then the inactivation slows down from the effect of pH on the steady state kinetics of the enzyme catalyse reaction, it has been concluded that a masked basic group (pKa = 6.9) is present at the active site. Succinate has been found to exhibit specific interaction with the enzyme, which is characterised by spectral effect. The enzyme possesses to type of succinate binding refered to as the "tight" and the "loose" sites. It has been suggested that "loose" sites may actually represent the "regulatory" sites of the enzyme. ICL has been shown to be a metalloenzyme the thermal inactivation of native and apoenzyme, binding of succinate and Mg 2+ to the apo-enzyme are suggestive of asymmetry in tetrameric enzyme molecule. It has been concluded that enzyme has a pre -existent asymmetry, which is an inherent property of the proteins and is not induce by any substrate or effector.

• Awarded CERTIFICATE OF MERIT along with a cash prize of Rs 75/- from Faculty of Science, BHU for securing FIRST RANK in M.Sc. (Biochemistry Previous) examination of Banaras Hindu University held in 1972.
• Chaired a Scientific Session of Workshop cum Symposium at Jodhpur, 1988.
• Served as Resource person to deliver Lecture in Academic Staff College, BHU, 1999.
• Delivered a series of Lectures at A.P.S. University, Rewa as visiting fellow, 2000.
• Delivered a series of Lectures at Biotechnology Centre, Allahabad University, 2001.
• Assigned substantial number of Lectures in Post Graduate Teaching Programme in Chemistry, Biotechnology, Applied Microbiology and Home Science for last several years.
• Awarded VIJAY RATTAN AWARD along with CERTIFICATE OF EXCELLENCE in 2005 by India International Friendship Society, New Delhi.
• Selected BEST CITIZENS OF INDIA AWARD in 2006 by Best Citizen Publishing House, New Delhi.
• Selected for RAJIV GANDHI SHIROMANI AWARD 2007 by India International Friendship Society, New Delhi.
• Selected for Shiksha Rattan Puraskar award , 2008 by India International Friendship Society, New Delhi.
• Appointed as Member of Academic Council, BHU , 2008-2011.

EXTRA CURRICULAR ACTIVITIES:

• Worked as Warden, Bhabha Hostel, BHU (1984-1987)
• Worked as Proctor in the Proctoral Board, BHU (1991-1992)
• Worked as Programme Officer (NSS) BHU (2002-2005).
• Worked as Administrative Warden of Broacha Hostel and Dr. C.P.R. Aiyer Hostel (2003-2006).
• Working as Observer in Central Vigilance Board, BHU for P.E.T. and other University Examinations.
• Working as University Representative to conduct MCA, MBA, RET, UET, PET tests at Delhi, Calcutta and Mumbai centers for several years.
• Elected as Treasurer, the University Club BHU, 2010.

TEACHING EXPERIENCE: 1977 To-date: Courses Taught:

• Organic Reaction Mechanism as applied to Biochemical Systems.
• Bioenergetics and Intermediary Metabolism of carbohydrates, proteins and amino acids, nucleic acid, lipids and fats.
• Isolation and purification of enzymes, kinetics and mechanism of enzyme action, regulation of enzyme activity and enzyme technology.

RESEARCH EXPERIENCE:

37 years: 1973-79:- Doctoral

1979 - to continue:- Post Doctoral

Ph.D. Thesis Title: Kinetics and Regulatory role of Isocitrate Lyase of Castor Endosperm

Four students have received Ph.D. under my supervision and one has submitted the thesis (Aug.,2010)

Five students are currently working under my supervision for their Ph.D. Degree.

Seminar/Symposia Organized


(i) UGC sponsored workshop on Biochemical Education, during October 24-28, 1985 (member local organizing committee).

(ii) International Seminar on Structure and Function of Enzymes, during Oct. 22-25, 1986 (member local organizing committee).

(iii) 4th Convention of The Indian Society of Agricultural Biochemists and Symposium on Recent Developments in Biochemistry, during March 20-21, 1995 (member local organizing committee).

(iv) International conference on "Role of Biomolecule in Food and Health Improvement"(ICBFH-2010) during Feb.17-20, 2010 (member local organizing committee).

I - Papers Presented in Symposium:

1. Kinetics and Regulatory role of isocitrate lyase from castor seedlings. Presented in Annual Convention of Chemists, 1977, at Jaipur, Abst. P.14.

2. Molecular asymmetry in isocitrate lyase of castor endosperm. Presented in Society of Biological Chemists, 1978, New Delhi, Abstract p. 153.

3. Possible active site groups and regulatory significance of isocitrate lyase of castor endosperm. Presented in Annual Convention of Chemists at Madras, 1981, abstract p.5.

4. Role of Mg2+ ions in the catalytic activity of isocitrate lyase. Presented in Indian Council of Chemists at Srinagar (J & K), 1982, Abstract p. 9.

5. Significance of SH group in the catalysis of isocitrate lyase of caster endosperm. Published in anuual convention of chemists at Calcutta, 1984, Abstract p.3.

6. Molecular Asymmetry and site : site heterogeneity in the action of isocitrate lyase. Presented and participated in All India Summer Institute in Biochemistry, held at Hyderabad, 1984.

7. Partcipated in Workshop on Biochemical Education, held at Banaras Hindu University, 1985.

8. Presented a paper in 73rd Indian Science Congress, held at Delhi, 1986.

9. Chemical reaction mechanism and active site groups of isocitrate lyase. Presented in International Seminar on Enzyme Structure and Function, held at BHU, 1986, Abstract p.15.

10. Studies on steady state kinetics and regulation of isocitrate lyase of castor endosperm. Presented Seminar on Enzyme Structure and Function, held at BHU, 1986, Abstract p.37.

11. Prartipated in the Workshop on Computer Aided Instructions in Chemistry, held at Banaras Hindu University, 1987.

12. Studies of NADP-Linked isocitrate dehydrogenase. Presented in 75th Indian Science Congress, held at Pune, 1988, abstract p. 58.

13. Participated & presented a paper in workshop cum Symposium on Polymer Aided Reactions, held at Jodhpur, 1988, abstract p. 41.

14. Isolated and charactesization of NADP-specific isocitrate dehydrogenase of germinating Mung beans. Presented in 75th Annual Meeting of SBC, held at Delhi, 1988, abstract no. 186.

15. Isolation and characterization of NADP+-specific isocitrate dehydrogenase. Presented in 25th Annual Convention of Chemists, held at Calcutta, 1988 abstract Bio. 6.

16. Participated in 1st National Workshop on High performance Liquid Chromatography, held at AIIMS, New Delhi, 1989.

17. Studies on Steady State kinetics of NADP+ isocitrate dehydrogenase from mung bean. Presented in 79th Indian Science Congress, held at Baroda, 1992.

18. Regulation of NADP+-linked isocitrate dehdrogenase of germinating mung bean. Presented in SBC, held at Madurai, 1993.

19. Molecular Symmetry in NADP+-specific isocitrate dehydrogenase of mung bean. Presented in 16th IUBMB, Satellite Meeting on Enzyme Mechanism, IISc, Bangalore, 1994.

20. Molecular Symmetry in germinating pea seeds NADP+ specific isocitrate dehydrogenase. Published in 17th International Congress of Biochemistry and Molecular Biology, 1997. Annual Meeting of the American Society of Biochemistry and Molecular Biology in San Francisco, California, USA.

21. Molecular Symmetry in germinating mung bean NADP+ specific isocitrate dehydrogenase. Presented in 67th Annual Meeting of SBC, 1998, held at New Delhi, Abstract p. 59.

22. Studies on NADP+ -linked isocitrate dehydrogenase in germinating urd bean seeds (Phaseolus mungo L). Presented in third symposium on current advances: Application in Health Environment and Agriculture at Lucknow (2007). Abstract p. 79.

23. Internal Transcribed Spacer 2 of Ribosomal DNA: A molecular marker for phylogenetic and population analyses in Mahseer. Presented in International Conference on Role of Biomolecule in Food Security and Health Improvement held at Banaras Hindu University (2010). p.139.

II- Journal Papers (Published/Accepted)


1. Malhotra, O.P., and Srivastava, P.K. (1982): Isolation and characterization of isocitrate lyase of caster endosperm. Archives of Biochemistry and Biophysics, 214, 164-171.

2. Malhotra, O.P., Dwivedi, U.N., and Srivastava, P.K. (1984): Steady state kinetics and negative cooperativity in the action of isocitrate lyase. Indian Journal of Biochemistry & Biophysics, 21, 99-105.

3. Malhotra, O.P., Srivastava, P.K., and Dwivedi, U.N. (1984): Role of metal ions in activity and site: site heterogeneity in isocitrate lyase of caster seedling endosperm. Archives of Biochemistry and Biophysics, 235, 612-617.

4. Malhotra, O.P., Dwivedi, U.N., Singh, J., and Srivastava, P.K. (1987): Chemical reaction mechanism and active site groups of isocitrate lyase. Indian Journal of Biochemistry & Biophysics, 24, 57-62.

5. Malhotra, O.P., Singh, J., and Srivastava, P.K. (1991): Significance of sulphydryl groups in the activity of isocitrate lyase of caster endosperm. Plant Science, 75, 47-53.

6. Srivastava, P.K., and Singh, D.S. (1999): Studies on NADP+ specific isocitrate dehydrogenase of germinating pea seeds. Journal of Scientific Research, 49, 117-129.

7. Srivastava, P.K., and Singh, D.S., (2001): Isolation and characterization of NADP+-linked isocitrate dehydrogenase of germinating pea seeds (Pisum sativum). Indian Journal of Biochemistry & Biophysics, 38, 335-341.

8. Srivastava, P.K., Pathak, K., and Prakash, Om (2004): A Study of NADP+-linked Isocitrate dehydrogenase from germinating mung bean (Vigna radiata). Journal of Plant Biochemistry & Biotechnology, 13, 145-148.

9. Srivastava, P.K., Pathak K., and Singh S. (2009): Identification of active site groups in mung bean NADP+-linked isocitrate dehydrogenase. Journal of Scientific Research. 53,63-71.

10. Sarkar, N., Srivastava, P.K., and Dubey, V.K (2009): Understanding the language of Vitamin C. Current Nutrition & Food Science 5, 1-3.

11. Srivastava, P.K., Pathak. K., and Singh S. (2010): Kinetics of Inactivation of NADP+-Linked Isocitrate Dehydrogenase from Germinating Mung Bean (Vigna radiata). Journal of Plant Biochemistry & Biotechnology 19, 251-254.

12. Srivastava, P.K., Srivastava, G. K., and Singh S. (2010): Isolation and characterization of NADP+ -linked Isocitrate dehydrogenase in germinating urd bean seeds (Phaseolus mungo). Journal of proteins and Proteomics 1, 3-10.

13. Srivastava, P.K., Srivastava, G.K., Mani, I., Yadav, S., Anand, A. (2011): Role of metabolites and significance of SH groups im the action of NADP+ linked isocitrate dehydrogenase of Urdbean Seeds (Phaseolus mungo L.). Asian journal of biochemistry 6 (2): 181-190.

14. Mani, I., Kumar, R., Singh, M., Kushwaha, B., Nagpure, N.S., Srivastava, P.K., Murmu, K., Rao, D.S.K., and Lakra, W.S. (2009): Karyotypic diversity and evolution of seven mahseer species (Cyprinidae) from Indian Journal of Fish Biology.75, 1079-1091.

15. Mani, I., Kumar, R., Kushwaha, B., Singh, M., Nagpure, N.S., Srivastava, P.K., Lakra, W.S. (2010): Cytogenetic characterization of an endemic mahseer, Tor mosal mahanadicus (David, 1953; Teleostei : Cyprinidae). The Nucleus, 53, Issue 3, 109-114.

16. Mani, I., Kumar, R., Singh, M., Nagpure, N.S., Kushwaha, B., Srivastava, P.K., Rao, D.S., Lakra, W.S. (2011): Nucleotide variation and physical mapping of ribosomal genes using FISH in genus Tor (Pisces, Cyprinidae). Mol Biol Rep. 38(4):2637-47.

17. Mani, I., Kumar, R., Singh, M., Kushwaha, B., Nagpure, N.S., Srivastava, P.K., Lakra, W.S. (2013): Chromosomal distribution of constitutive hetrochromatin in eight species of mahseer (Family Cyprinidae) from India. Indian Journal of Biotechnology 12, 178-186.

18. Yadav, S., Singhal, N.K., Singh, V., Rastogi, N., Srivastava, P.K. and Singh M.P.(2009): Association of single nucleotide polymorphism in CYP1B1 and COMT genes with breast cancer susceptibility in Indian women. Disease Markers, 27, 1-8.

19. Yadav, S., Gupta, S.P., Srivastava, G., Srivastava, P.K., Singh, M.P. (2012): Role of secondary mediators in caffeine-mediated neuroprotection in maneb- and paraquat-induced Parkinson's disease phenotype in the mouse. Neurochem Res. 37(4):875-84.

20. Yadav, S., Dixit, A., Agrawal, S., Singh, A., Srivastava, G., Singh, A.K., Srivastava, P.K., Prakash, O., Singh, M.P. (2012): Rodent models and contemporary molecular techniques: notable feats yet incomplete explanations of Parkinson's disease pathogenesis. Mol Neurobiol. 46(2):495-512.

21. Srivastava, A.K., Ali, W., Singh, R., Bhui, K., Tyagi, S., Al-Khedhairy, A.A., Srivastava, P.K., Musarrat, J., Shukla, Y. (2012): Mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytees. Life Science 90; (21-22): 815-24.

22. Srivastava, A.K., Srivastava, P.K., Al-Khedhairy, A.A., Musarrat, J., Shukla, Y. (2012): Allethrin-induced genotoxicity and oxidative stress in Swiss albino mice. Mutation Research 30; 747(1):22-8.

23. Srivastava, A.K., Mishra, S., Ali, W., Srivastava, P.K., Shukla, Y. (2013): Mancozeb-induced genotoxicity, cytotoxicity, in exposed agriculture workers. Toxicology and Applied Pharmacology (Communicated).

24. Anand, A., Srivastava, P.K. (2012): A molecular description of acid phosphatase. Appl Biochem Biotechnol 167; (8): 2174-97.

25. Srivastava, P.K., Anand, A. (2014): Immobilization of acid phosphatase from Vigna aconitifolia seeds on chitosan beads and its characterization. Int J Biol Macromol 64:150-4.

26. Anand, A., Srivastava, P.K. (2014): Isolation and enzymatic properties of a nonspecific acid phosphatase from Vigna aconitifolia seeds. Biotechnology and Applied Biochemistry 61(2), 145�152.

27. Srivastava, P.K., Anand, A. (2015): The inhibitory effect of metals and other ions on Acid Phosphatase activity from Vigna aconitifolia Seeds. Prep Biochem Biotechnol 2; 45(1):33-41.

28. Singh, S., Anand, A., Srivastava, P.K. (2012): Regulation and properties of glucose-6-phosphate dehydrogenase a review. International Journal of plant Physiology and Biochemistry 4; (1), 1-19.

29. Gupta, V.K., Pal, A., Singh, A., Srivastava, P.K., Darokar, M.P. Punica granatam root ethanol extract inhibit growth of clinical isolates of b- lactamase producing methicillin resistant Staphylococcus aureus by altering cell membrane permeability (2012). International journal of food properties (Accepted).

30. Gupta, V.K., Verma, S., Gupta, S., Singh, A., Pal, A., Srivastava, S.K., Srivastava, P.K., Singh, S.C., Darokar, M.P. (2013): Membrane-damaging potential of natural L-(-)-usnic acid in Staphylococcus aureus. Eur J Clin Microbiol Infect Dis 31; (12):3375-83.

31. Gupta, V.K., Verma, S., Pal, A., Srivastava, S.K., Srivastava, P.K., Darokar, M.P. (2013): In vivo efficacy and synergistic interaction of 16α-hydroxycleroda-3, 13 (14) Z-dien-15, 16-olide, a clerodane diterpene from Polyalthia longifolia against methicillin-resistant Staphylococcus aureus. Appl Microbiol Biotechnol 97(20); 9121-31.