Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake
Hazaa, M., Sabae, S., T, A., Eldourghamy, A., Sayed, H. (2018). Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake. EKB Journal Management System, 5(3), 162-173. doi: 10.21608/jbes.2018.370465
Mahmoud M. Hazaa; Shawky Z. Sabae; Areej Ibrahim F. T; Ayman S. Eldourghamy; H. Sayed. "Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake". EKB Journal Management System, 5, 3, 2018, 162-173. doi: 10.21608/jbes.2018.370465
Hazaa, M., Sabae, S., T, A., Eldourghamy, A., Sayed, H. (2018). 'Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake', EKB Journal Management System, 5(3), pp. 162-173. doi: 10.21608/jbes.2018.370465
Hazaa, M., Sabae, S., T, A., Eldourghamy, A., Sayed, H. Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake. EKB Journal Management System, 2018; 5(3): 162-173. doi: 10.21608/jbes.2018.370465

Production, Purification and Characterization of Alfa-amylase produced by Bacteria from Pharaonic Lake

Journal of Basic and Environmental Sciences
Volume 5, Issue 3, July 2018, Pages 162-173    PDF (251.98 K)
Document Type: Original Article
DOI: 10.21608/jbes.2018.370465
Authors
Mahmoud M. Hazaa1; Shawky Z. Sabae2; Areej Ibrahim F. T3; Ayman S. Eldourghamy4; H. Sayed3
1Department of plant, Faculty of science, Benha University, Benha, Egypt
2National Institute of oceanography and fisheries, Egypt
3National Water Research Centre, El-Qanater El- Khayreia, Egypt
44 Env. Biotech. Dept. Gen. Eng. Biotech. Res. Ins. University of Sadat City
Abstract
In the present study thirty bacteria isolates collected from Pharaonic lake, Menoufiya Governorate, Egypt. Out
of which 12 isolates that were α-amylase producer. Isolate No. 1 was selected as the best enzyme producer,
identified as Alloiococcus otitis and was used for further studies. Attempts were made to optimize the cultural
condition for getting high yields of enzyme. The optimum level of pH for enzyme production was 8.0 on Bennett
medium after 8 hours. Among carbon sources, maltose supported maximum production of α-amylase followed by
lactose, sucrose, starch and manitol. Urea was the best nitrogen source for enzyme production, followed by L- asparagine. The purified fraction detected by the appearance of a single band corresponding to 38 k Da in SDS- PAGE, and the enzyme was purified 67.0-fold with amylase units 225 ml and specific activity of 234.4 U/mg.
The pure enzyme was stable at all degrees of temperatures (20–70ºC). The maximum activity was at pH of 9.0.
Starch was the best substrate. CaCl2, MgCl2 and KCl were the best activators. There is a continuous decreasing in
the activity due to the increase of SDS and H2O2 concentrations. Enzyme was stable in presence of butanol, n- hexane and toluene.
Keywords
α-amylase; Bacteria; Purification
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