Production, characterization and immobilization of Aspergillus versicolor L-asparaginase onto multi-walled carbon nanotubes
Ahmed A. Haroun, Hanan Haj Ahmad, Abdel‐Tawab H. Mossa, Samia M.M. Mohafrash, Eman F. Ahmed, E Kelo, T Noronkoski, I Mononen, N Verma, K Kumar, G Kaur, S Anand, A Shrivastava, A Khan, M Khurshid, M Kalam, S Jain, P Singhal, Z Moola, M Scawen, T Atkinson, D Nicholls, A Soares, G Guimares, P Bronislaw, R Pitombo, J Abraho-Neto, G Baskar, R Aiswarya, S Renganathan, M Roy, V Das, A Patra, N El-Naggar, S Deraz, S El-Ewasy, G Suddek, M Zolfaghar, M Amoozegar, K Khajeh, H Babavalian, H Tebyanian, F Qeshmi, A Homaei, P Fernandes, S Javadpour, V Avamis, E Panosyan, A Klibanov, Y Zhang, M Tao, W Shen, Y Zhou, Y Ding, M Yan, W Zhou, J Jean-Francois, E D'urso, G Fortier, M Gaspar, R Perez-Soler, M Cruz, T Hilder, J Hill, Q Husain, I Pavlidis, T Tsoufis, A Enotiadis, D Gournis, H Stamatis, S Shah, K Solanki, M Gupta, P Zhang, D Henthorn, N Mubarak, J Wong, K Tan, J Sahu, E Abdullah, N Jayakumar, P Ganesan, E Khaled, M Kamel, H Hassan, A Haroun, A Youssef, H Aboul-Enein, H Amin, A Haroun, A Haroun, A Mossa, S Mohafrash, A Haroun, H Amin, Abd El-Alim, S Preparation, A Haroun, H Amin, Abd El-Alim
Abstract
The goal of this work is to study the optimum production of L-ASNase and the influence of oxidized multi-walled carbon nanotubes (ox-MWCNTs) on the biological activity of the isolated L-ASNase. Isolation and production of L-ASNase from different screened fungi in the presence of L-asparagine substrate were carried out. The isolated enzyme was identified by molecular18S rRNA analysis. The partial purified L-ASNase having 6.77 purification fold was immobilized onto ox-MWCNTs using physical adsorption technique with maximum yield about 54.4%. In vitro cytotoxicity of L-ASNase against normal fibroblast cell line (BHK-21) was examined relative to the immobilized one using SRB assay. Also, in vivo biological examination using liver and kidney dysfunction biomarkers in the treated mice (AST, ALT, LDH, lipase and α-amylase) was also investigated. Among the ten screened fungi, Aspergillus versicolor was the most potent one with maximum activity about 45.5 U/mL. The results showed that the most appropriate production condition was at asparagine concentration of about 0.04 M and pH 8. The immobilized L-ASNase retained a complete activity (100%) after 30 min at 45ᴼC of incubation relative to that in case of the free one. The toxicity of L-ASNase was significantly decreased up to 50 μg/mL after immobilization onto MWCNTs. Besides, the immobilized L-ASNase exhibited good storage ability and stability at high temperature relative to that in case of the free one.