Agro Waste Utilization for Cost-Effective Production of l-Asparaginase by Pseudomonas plecoglossicida RS1 with Anticancer and Acrylamide Mitigation Potential

Abstract

Agricultural wastes such as the peels of onion and garlic were used as a supplement along with l-asparagine for the very first time to produce increased yield of l-asparaginase by Pseudomonas plecoglossicida RS1. Statistical optimization strategies such as response surface methodology were used to generate a medium composition containing extracts of 0.9 (v/v) of garlic peel waste and 0.5% (v/v) onion peel waste along with 0.2% (w/w) l-asparagine, which yielded a twofold increase in the enzyme activity compared to the unsupplemented minimal (M-9) medium. The presence of l-asparagine content in the peel extract was confirmed by high-performance liquid chromatography. Further, l-asparaginase was purified to homogeneity, and identity was confirmed by matrix-assisted laser desorption ionization time-of-flight analysis. The application of the purified l-asparaginase as a therapeutic was studied in HeLa cells which showed a p53-mediated G2 cell cycle arrest. Moreover, the purified l-asparaginase showed effective acrylamide mitigation in vitro, at 6 IU, and its effective degradation was also demonstrated by the effect on chemotactic index of Caenorhabditis elegans and the restoration of the cognitive abilities of C. elegans which was coexposed to acrylamide and l-asparaginase compared to that exposed to acrylamide alone. Thus, l-asparaginase, with multipotent applications, was produced by effective waste utilization for economical commercial production.

Production, characterization and techno-economic evaluation of Aspergillus fusant l-asparaginase

“…They cause intracellular depletion of serum l -asparagine leading to general impairment in glutamine metabolism through the tricarboxylic acid cycle, thus interfering with the ability of lymphoblasts to take up the amino acid (Vala et al 2018 ). Additionally, they are involved in the prevention of other kinds of cancers through scavenging of free-radicals and acrylamide reduction in fried, roasted or baked foods (Shakambari et al 2017 ). They are produced naturally by all biological systems, typically plants and microorganisms, and could be expressed in biotechnologically-safe heterologous hosts (Mihooliya et al 2020 ).…”

Antitumor activity and ability to prevent acrylamide formation in fried foods of asparaginase from soybean root nodules

“…It is one of the major drugs used in the treatment of acute lymphoblastic leukemia (ALL) and other related blood cancers (Duval et al, ; Krishnapura & Belur, ). L‐asparaginase is also used in food industry to prevent the formation of acrylamide in oven‐heated and fried foods where the reaction of free asparagine and reducing sugars gives rise to acrylamide which is a carcinogen (Shakambari, Kumar, Ashokkumar, & Varalakshmi, ). Although L‐asparaginases from Escherichia coli and Erwinia chrysanthemi are commercially available for the treatment of ALL, their therapeutic utilization is impeded by untoward complications involving hypersensitivity and several other toxic side effects (Krishnapura et al, ; Pieters et al, ).…”

Produção de L-asparaginase por >i<Escherichia coli>/i< BL21 (DE3) recombinante em meio de cultivo proveniente de fonte alternativa de carbono

“…A atual dependência da importação da L-asparaginase impulsiona o desenvolvimento biotecnológico nacional para a obtenção de um processo com moléculas eficientes e bioprocessos industrialmente viáveis (técnica e economicamente). Sua produção pode ter processos otimizados com o uso de microrganismos geneticamente modificados, onde é possível obter enzimas com propriedades melhoradas (SANTOS, 2017) e com uso de matérias-primas renováveis (SHAKAMBARI et al, 2017). Os avanços tecnológicos nessa área também podem proporcionar um crescimento de produtividade podendo adotar estratégias de produção mais flexíveis e desta forma, incentivar o desenvolvimento biotecnológico nacional.…”