Isolation, Optimization, and Antitumor Activity of L-Asparaginase Extracted From Pectobacterium Carotovorum and Serratia Marcescens on Human Breast Adenocarcinoma and Human Hepatocellular Carcinoma Cancer Cell Lines

Abstract

Objectives: The objective of this research was to obtain isolates capable of producing a high yield of L-asparaginase enzyme and to evaluate the antitumor activity of the purified enzyme against different cancer and normal cell lines.
 Methods: Isolation of bacteria was performed by the serial dilution technique of soil samples collected from Cairo, Egypt, using modified M9 agar plates. Culture filtrates of selected isolates were quantitatively screened for L-asparaginase production using well-diffusion and direct nesslerization techniques. Factors influencing L-asparaginase activity were optimized by studying the effect of physical and nutritional conditions on the enzyme activity. The purification of L-asparaginase extracted from both the isolates was achieved using chilled acetone (−20°C), followed by gel filtration on Sephadex G-100. The anticancer activity of the purified enzyme against human breast adenocarcinoma (MCF-7), human hepatocellular carcinoma (HepGII) and homo sapiens human (WISH) cell line was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay.
 Results: Two L-asparaginase producers were identified by Biolog identification system as Pectobacterium carotovorum and Serratia marcescens. Optimization increased the production of L-asparaginase to 4.835 and 5.221 U/ml for P. carotovorum and S. marcescens, respectively. L-asparaginase was extracted, purified, and tested in vitro for cytotoxic activity using 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT assay) against MCF-7, HepGII, and WISH cell line. L-asparaginase from P. carotovorum and S. marcescens was neutral to normal epithelial WISH cells. On the other hand, L-asparaginase from both isolates was cytotoxic to MCF-7 and HepGII cancer cell lines with an half maximal inhibitory concentration of 15 μg/ml and 26 μg/ml and 26 μg/ml and 25 μg/ml, respectively.
 Conclusion: L-asparaginase extracted from P. carotovorum and S. marcescens showed remarkable anticancer activity. Further studies on hypersensitivity action need to be carried out to recommend the use of L-asparaginase as an alternative to commercially available preparations.

Genomic analysis and plant growth-promoting potential of a Serratia marcescens isolated from food

“…Among some products, we can highlight the versatile enzyme L-asparaginase that includes the medical application of its antimicrobial and anti-inflammatory properties besides its use as a chemotherapeutic agent in different cancer cell lines (Abdel-Razik et al, 2019). The strategy of using novel therapeutic agents isolated from S. marcescens has been previously demonstrated in a marine strain with antioxidant and antibacterial activity against some Gram-positive and Gram-negative bacteria.…”

“…Both disease and therapy are present in the dual role of S. marcescens in the clinic, responsible for outbreaks (Cristina et al, 2019) was even identified as an opportunist in outbreaks during covid-19 treatment (Amarsy et al, 2020). However, it has shown an extensive presence of enzymes with antimicrobial and anti-inflammatory properties proving effective to even resistant microorganisms and with the possibility of use as a chemotherapeutic agent in different cancer cell lines (Abdel-Razik et al, 2019;Pavithrra & Rajasekaran, 2020).…”