roduction of Chemotherapeutic Agent L-asparaginase from Gamma-Irradiated Pseudomonas aeruginosa WCHPA075019.

Abstract

Because of the dangers and painful effects of chemotherapeutic drugs, the need for therapeutic agents with less adverse effects will increase several times in the coming years. L-asparaginase enzyme is an effective antitumor agent, especially acute lymphoblastic leukemia, with no side effects compared to other chemotherapeutic agents. Microorganisms are emerging as a safer source of L-asparaginase. Therefore, the findings of new L-asparaginase-producing bacterial strains with high yield for therapeutic applications become necessary. From twenty bacterial isolates tested for their L-asparaginase activity, 16S rRNA sequencing for the most potent isolate showed that the selected isolate had 100% identity to pseudomonas aeruginosa strain (accession number: WCHPA075019). In the presence of L-asparagine (1%) and glucose (1%) as nitrogen and carbon sources at a low dose of gamma radiation (0.75 kGy), the maximum productivity of Lasparaginase was reached after 2 days at 35 ° C, pH 7.6 under shaking at 200 rpm. Purification of L-asparaginase with 70% ammonium sulphate, followed by Sephadex G100 increases enzyme purity by 1.5-fold after gel filtration. Pure Lasparaginase had a molecular weight of 123 kDa by SDS-PAGE. The maximum activity of the enzyme against L-asparagine was detected at 35°C and pH 9.0 after 30 min and 200 mM substrate. L-asparaginase activated in the presence of metal ions such as K+, and Na+, not affected when exposed to EDTA and strongly inhibited in the presence of Ba2+, and Cd2+. The anticancer activity of the purified enzyme was tested in vitro against three types of cell line carcinoma. The growth inhibition of L-asparaginase for HEPG2 carcinoma cell line (IC50 value of 3.5 µg/ml) was greater than the inhibition of HCT and MCF-7 carcinoma cell lines with IC50 value of 3.8 and 12.5 µg/ml, respectively relative to the growth of the untreated control cells.

Production, characterization, and applications of a novel thermo-acidophilic L-asparaginase of Pseudomonas aeruginosa CSPS4

“…Nutrient source Sucrose (3%) and peptone (1%) showed maximum L-asparaginase production at 37 °C under acidic pH 6.0 after 48 hrs of incubation (Table 1). Very similar parameters were reported to produce L-asparaginase from another P. aeruginosa strain, where 1% (w/v) asparagine and glucose were reported for the maximum productivity after 48 hrs incubation at 35 °C (Amany et al 2021). Statistical optimization using PB and Response Surface Methodology (RSM) is always beneficial to understand the interaction among the physical and chemical parameters for enhancing the production of enzymes (Sharma and Satyanarayana 2011;Joshi and Satyanarayana 2015;Talluri et al 2019).…”

“…Comparable properties in another L-asparaginase (Topt: 60 °C and pHopt : 5.6) were also observed; however, it was from a fungal strain Aspergillus tubingensis SY1 (Yahya et al 2016). Most of the Pseudomonas-derived L-asparaginases exhibit optimum activity at neutral pH (Fatima et al 2019;Amany et al 2021;Kambe et al 2012). Alkaline L-asparaginases have also been reported from various species of Pseudomonas (Qeshmi et al 2022;EI-Bessoumy et al 2004;Kumar et al 2022).…”

“…It was consistent with the recombinant L-asparaginase of P. aeruginosa CSPS4 (Kumar et al 2024). However, L-asparaginase of P. aeruginosa WCHPA075019 (Amany et al 2021) and P. aeruginosa 50071 (EI-Bessoumy et al 2004) showed a higher molecular weight L-asparaginases of 123 kDa and 160 kDa, respectively. Kumar et al (2022) also reported a high molecular mass L-asparaginase of 148.0 kDa from Pseudomonas sp.…”

“…Therefore, pharmaceutical industries demand glutaminase-free L-asparaginase enzymes. 2021) reported several L-asparaginase producers from fish, meat, and egg sources (Amany et al 2021).…”

Caracterización bioinformática y producción de L-asparaginasa de Bacillus sp. M62 aislado de las salinas de Maras, Cusco, Perú

“…M62, debido a la mayor actividad L-asparaginasa obtenida en comparación con L-arginina, NaNO 3 y (NH 4 ) 2 SO 4 en otras cepas de Bacillus (Rahimzadeh et al 2016). Su efecto como inductor de L-asparaginasa se verificó en P. aeruginosa WCHPA075019 con la obtención de 170.70 U/mg, superior a lo obtenido con extrac-to de levadura, peptona, glutamina y arginina (Amany et al 2021). También, Jetti et al (2017), reportaron como significativo el uso de L-asparagina en Pectibacterium carotovorum MTCC 1428, por la cual se obtuvo hasta 26 U/mL.…”