Evaluation of antitumor activity of fungal L-glutaminase produced by egyptian isolates
Mary S. Khalil, Hani Moubasher, Mokhtar El-Zawahry, Marina Michel, A Jemal, R Siegel, E Ward, Y Hao, J Xu, T Murray, M Thun, A Jemal, F Bray, M Center, J Ferlay, E Forman, G Samjotha, I Sudheer, G Pasut, M Sergi, F Veronese, C Langer, J Harris, E Horwitz, N Nicolaou, M Kies, W Curran, S Wong, K Ang, O Darwesh, M Eida, I Matter, A Sabu, M Chandrasekaran, A Pandey, W Criss, M Eida, O Darwesh, I Matter, D Wise, C Thompson, K Sarada, A Sabu, K Nampoothiri, A Pandey, R Balagurunathan, M Radhakrishnan, S Somasundaram, Z Kheiralla, M Hewedy, H Mohammed, O Darwesh, K Yoshimune, D Marrez, A Abdelhamid, O Darwesh, M Hasanin, O Darwesh, I Matter, H El-Saied, R Gulati, R Saxena, R Gupta, O Darwesh, S El-Maraghy, H Abdel-Rahman, R Zaghloul, G Nagendra-Prabhu, M Chandrasekaran, O Lowery, A Rosebrouph, N Farr, M Bradford, K Barakat, S Hassan, O Darwesh, H Hussein, O Darwesh, B Mekki, S El-Hallouty, O Darwesh, I Matter, M Eida, T Mosmann, A Porter, R Janicke, G Koopman, H Hussein, O Darwesh, B Mekki, K Nathiya, S Sooraj Nath, J Angayarkanni, M Palaniswamy, R Shimaa, S Raed, I Elshahawy, H Abouelnasr, S Lashin, O Darwesh
Abstract
L-glutaminase is an amidohydrolase that catalyzes L-glutamine to L-glutamic acid and has increased impressive consideration due to its potential anticancer activity. The objective of this work was to isolate different fungi that produce L-glutaminase which has antitumor activities. Three fungal species that were isolated on the selective medium showed L-glutaminase activity. The species were identified as a Penicillium crustosum, Emericella nidulans and Mucor circinelloides. Mucor circinelloides showed potential L-glutaminase production (5.16 U/ml). Different pH, L-glutamine concentrations, temperatures, and incubation periods were carried out. The most enzyme production was shown at pH 8, temperature 30 ºC, with 0.6% of the L- glutamine. L-glutaminase was extracted and purified with gel filtration and ion exchange (DEAE Sephadex A50). The purified enzyme was found to possess a specific activity 1.55 U/mg and was monitored by SDS-PAGE with molecular weight 70 kDa. Purified L-glutaminase exhibited cytotoxic activity against HepG2 and PC3 cell lines with Ic50 value 0.067 mg/ml and 0.079 mg/ml respectively. Caspase 3 colorimetric assay and flow cytometry was done to detect the activation of apoptosis among the enzyme. The enzyme showed positive results on both types of cell lines.