Purification, Characterization of L-Methioninase fromCandida tropicalis, and Its Application as an Anticancer

Abstract

The aim of the present study is to purify L-methioninase from Candida tropicalis 34.19-fold with 27.98% recovery after ion exchange chromatography followed by gel filtration. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular weight of 46 KDa. Its optimum temperature was 45 to 55 and thermal stability was 55°C for 15 min. The enzyme had optimum pH at 6.5 and stability at a pH range of 5.5 to 7.0 for 24 hr. The maximum activity was observed with substrate concentration of 30 µM and Km was 0.5 mM. The enzyme was strongly inhibited by Cd+2 and Cu+2 while it was enhanced by Na+, Ni+2, and Mg+2 at 10 mM while Ca+2 had slight activation at 20 mM. In addition, the potential application of the L-methioninase as an anticancer agent against various types of tumor cell lines is discussed.

Purification and Characterization of New Alkaline L-methioninase from Aspergillus ustus AUMC 1051 Grown under Solid-State Fermentation Conditions.

“…On the other hand, the enzyme activity was inhibited in the presence of Cu +2 , Zn +2 , Hg +2 and Fe +2 . These results are in consistent with that reported for fungal L-methioninase (Abu-Seidah and Youssef, 2000, El-Sayed, 2009and Selim et al, 2015. Generally, the stimulatory effect of metal ions may be ascribed to their stabilizing effect to the conformational structure, protection and Guijarro, 2001).…”

“…These results are in agreement with the previous findings on other L-methioninase from Clostridium sporogenes (Kreis and Hession, 1973), Cladosporium cladosporioides (Abu-Seidah and Youssef, 2000) and A. flavipes (El-Sayed, 2009). While, our results are in disagreement with that obtained from Candida tropicalis (Selim et al, 2015). The enzyme had a good stability on the alkaline side.…”

“…The appearance of L-methionine as a single band ensures the homogeneity and purity of the enzyme. Coincident with our results, the molecular weight of the purified L-methioninase produced by various bacterial (Nakayama et al, 1984;Dias andWeimer, 1998 andTakakura et al, 2006), protozoal (Tokoro et al, 2003) and fungal species (El-Sayed, 2009 andSelim et al, 2015) was in the range of 44 to 48 kDa.…”

“…It also retained about 55% of this activity at 50 o C and 35% at 60 o C after heating for complete hour. These results reveal that this enzyme from A. ustus was less thermostable than those of Candida tropicalis (Selim et al, 2015) but more stable than that of A. flavipes (El-Sayed, 2011). The purified enzyme preparation was found stable for six days when stored at 4 o C. T ½ of enzyme activity was found to be four weeks.…”

Antibacterial and Anticancer Activity and Untargeted Secondary Metabolite Profiling of Crude Bacterial Endophyte Extracts from Crinum macowanii Baker Leaves

“…P. putida TJ151 is able to produce fluorouracil which is a bioactive aromatic compound and it is an anticancer drug [ 59 ]. L-methioninase, an enzyme produced by Pseudomonas putida, has shown anticancer activity against leukemia cell lines, liver HepG2, breast MCF-7, lung A549, prostate PC3, and colon HCT116 [ 88 , 89 ]. Methioninase from P. putida and 5-fluorouracil work synergistically to inhibit tumor growth and hence the activity observed [ 90 ].…”