PRODUCTION, OPTIMIZATION AND PURIFICATION OF L-METHIONINASE FROM Aspergillus flavipes AUMC 1201
HAMED M. EL-SHORA *
Botany Department, Faculty of Science, Mansoura University, Egypt.
ALAA M. ABU-ZIED
Botany Department, Faculty of Science, Tanta University, Egypt.
OMYMA A. AWADALLA
Botany Department, Faculty of Science, Tanta University, Egypt.
SALLY M. METWALLY
Botany Department, Faculty of Science, Tanta University, Egypt.
NESSMA A. EL-ZAWAWY
Botany Department, Faculty of Science, Tanta University, Egypt.
*Author to whom correspondence should be addressed.
Abstract
L-methioninase (L-methionine-α-deamino-c-mercaptome-thane-lyase) (E.C 4.4.1.11) had received much attention for its importance as a drug target for many infectious diseases and for therapeutic interventions of various cancers. Therefore, the present study aimed to produce, optimize and purify extracellular L-methioninase from Aspergillus flavipes AUMC 1201. Enhanced production of L-methioninase occurred on the addition of 1.5% (w/v) glucose, 0.5% (w/v) L-methionine. Its optimum temperature was 30°C and the optimum pH was 8.0 for 8 days of shaking fermentation at 130 rpm. Purification fold of L-methioninase was 14.5 with 37.1% recovery after ammonium sulphate precipitation (90%), ion exchange chromatography on DEAE cellulose followed by gel filtration on Sephadex G-200. The purified enzyme revealed a single band on SDS-PAGE electrophoresis with a molecular weight of 48 KDa. Overall, purified L-methioninase can be used for further clinical applications.
Keywords: Aspergillus flavipes, L-methioninase, optimization, purification.