Purification, Characterization and Kinetic Properties of Penicillium cyclopium L-Asparaginase: Impact of Lasparaginase on Acrylamide Content in Potato Products and its Cytotoxic Activity

Order of Publishing in Issue: 
4
Volume :9
Issue :2
April, 2015
Page No: 
132-140
Authors: 
Mona S. Shafei[1], Heba A. El-Refai[1*], Hanan Mostafa [1], Abdel-Monem H.El-Refai[1], Fawkia M. El-Beih[2], Saadia M.Easa[2] and Sanaa.K.Gomaa[1]
Address: 
[1]Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo 12622, Egypt.
Address: 
[2]Ein-Shams University, Faculty of Science, Microbiology Department, Cairo, Egypt.
Email-ID: 
dr.heba_ar@yahoo.co

The present study was carried out for the purification and characterization of Lasparaginase produced by Penicillium cyclopium. Both food and medical applications were also carried in the present study. The molecular weight of enzyme was approximately 55 kDa as estimated by SDS-PAGE. The optimum temperature and pH of the enzyme were 37 ºC and 8, respectively with Vmax of 3333U/mg and Km of 0.3 mM. Studies for the substrate specificity indicated that, L-asparaginase has greater affinity towards L-asparagine. Metal ions such as Ca+2, Fe+3, Mn+2 and Hg+2 significantly affected the enzyme activity whereas presence of KCl and NaCl stimulated the enzyme activity. The impact of L-asparaginase on the acrylamide content reduction after high heat treatment in a model system as well as in potato based material was investigated. Addition of partial purified Lasparaginase enzyme followed by incubation of the mixture at 37 °C for 30 min led to 92 % reduction of acrylamide content. It showed potential candidate for food industry to reduce acrylamide content in starchy fried food commodities. The purified enzyme also inhibited the growth of three human cell lines including hepatocellular carcinoma (Hep-G2), breast carcinoma (MCF-7) and prostate carcinoma (PC3) with IC50 values of 14 μg/ml, 12.5 μg/ml and 37ìg/ml, respectively.

Keywords: 
Cytotoxicity, Acrylamide, Lasparaginase, Penicillium cyclopium, Purification.
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