L-asparaginase from <i>Aspergillus oryzae</i> spp.: effects of production process and biochemical parameters

Cunha, Marília Crivelari da; Aguilar, Jessika Gonçalves dos Santos; Lindo, Santa Maria Del Rosário Orrillo; Castro, Ruann Janser Soares de; Sato, Hélia Harumi

doi:10.1080/10826068.2021.1931881

Cited by 10 publications

(3 citation statements)

References 44 publications

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“…The primary effect of asparaginase is to deplete asparagine rather than glutamine. The administration of asparaginase was effective in vivo , reducing asparagine concentration to 1 μmol/L ( 23 ). We observed a significant reduction in plasma glutamine concentration to 33.31 ± 28.28μmol/L.…”

Section: Resultsmentioning

confidence: 99%

Exploring the causal relationship between glutamine metabolism and leukemia risk: a Mendelian randomization and LC-MS/MS analysis

Li,

Wang,

Zhang

et al. 2024

Front. Immunol.

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ObjectiveThis investigation sought to delineate the causal nexus between plasma glutamine concentrations and leukemia susceptibility utilizing bidirectional Mendelian Randomization (MR) analysis and to elucidate the metabolic ramifications of asparaginase therapy on glutamine dynamics in leukemia patients.MethodsA bidirectional two-sample MR framework was implemented, leveraging genetic variants as instrumental variables from extensive genome-wide association studies (GWAS) tailored to populations of European descent. Glutamine quantification was executed through a rigorously validated Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) protocol. Comparative analyses of glutamine levels were conducted across leukemia patients versus healthy controls, pre- and post-asparaginase administration. Statistical evaluations employed inverse variance weighted (IVW) models, MR-Egger regression, and sensitivity tests addressing pleiotropy and heterogeneity.ResultsThe MR findings underscored a significant inverse association between glutamine levels and leukemia risk (IVW p = 0.03558833), positing lower glutamine levels as a contributory factor to heightened leukemia susceptibility. Conversely, the analysis disclosed no substantive causal impact of leukemia on glutamine modulation (IVW p = 0.9694758). Notably, post-asparaginase treatment, a marked decrement in plasma glutamine concentrations was observed in patients (p = 0.0068), underlining the profound metabolic influence of the therapeutic regimen.ConclusionThis study corroborates the hypothesized inverse relationship between plasma glutamine levels and leukemia risk, enhancing our understanding of glutamine’s role in leukemia pathophysiology. The pronounced reduction in glutamine levels following asparaginase intervention highlights the critical need for meticulous metabolic monitoring to refine therapeutic efficacy and optimize patient management in clinical oncology. These insights pave the way for more tailored and efficacious treatment modalities in the realm of personalized medicine.

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Section: Resultsmentioning

confidence: 99%

Exploring the causal relationship between glutamine metabolism and leukemia risk: a Mendelian randomization and LC-MS/MS analysis

Li,

Wang,

Zhang

et al. 2024

Front. Immunol.

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“…The preference for SmF by Fusant-06 to produce l -ASNase was not surprising because wild type strains preferentially produced 6-times more l -ASNase in SmF than SSF (Ekpenyong et al 2021a ; unpublished data). Industrial production of l -ASNase has been mostly performed by SmF mainly from the point of view of higher yield (da Cunha et al 2022 ). In the 96-h fermentation in this study, SmF significantly outperformed ( p < 0.0001) SSF in all tested strains.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2023

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Protoplast fusion is one of the most reliable methods of introducing desirable traits into industrially-promising fungal strains. It harnesses the entire genomic repertoire of fusing microorganisms by routing the natural barrier and genetic incompatibility between them. In the present study, the axenic culture of a thermo-halotolerant strain of Aspergillus candidus (Asp-C) produced an anti-leukemic l-asparaginase (l-ASNase) while a xylan-degrading strain of Aspergillus sydowii (Asp-S) produced the acrylamide-reduction type. Protoplast fusion of the wild strains generated Fusant-06 with improved anti-leukemic and acrylamide reduction potentials. Submerged fed-batch fermentation was preferred to batch and continuous modes on the basis of impressive techno-economics. Fusant-06 l-ASNase was purified by PEG/Na+ citrate aqueous two-phase system (ATPS) to 146.21-fold and global sensitivity analysis report revealed polymer molecular weight and citrate concentration as major determinants of yield and purification factor, respectively. The enzyme was characterized by molecular weight, amino acid profile, activity and stability to chemical agents. Michaelis–Menten kinetics, evaluated under optimum conditions gave Km, Vmax, Kcat, and Kcat/Km as 6.67 × 10–5 M, 1666.67 µmolmin−1 mg−1 protein, 3.88 × 104 min−1 and 5.81 × 108 M−1.min−1 respectively. In-vitro cytotoxicity of HL-60 cell lines by Fusant-06 l-ASNase improved significantly from their respective wild strains. Stability of Fusant-06 l-ASNase over a wide range of pH, temperature and NaCl concentration, coupled with its micromolar Km value, confers commercial and therapeutic value on the product. Free-radical scavenging and acrylamide reduction activities were intermediate and the conferred thermo-halo-stability could be exploited for sustainable clinical and food industry applications.

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“…Therefore, more thermostable enzymes are required for this application. Approved by the FDA, commercial enzyme preparations from the mold fungi Aspergillus oryzae and Aspergillus niger (Acrylaway ® and PreventAse ® ) are currently used in the industry [7,8]. Acrylaway ® was used to treat and reduce acrylamide formation in coffee beans [9].…”

Section: Introductionmentioning

confidence: 99%

Thermal Inactivation Mechanism and Structural Features Providing Enhanced Thermal Stability of Hyperthermophilic Thermococcus sibiricus L-Asparaginase in Comparison with Mesophilic and Thermophilic L-Asparaginases

et al. 2023

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This work aimed to study the structural features and mechanisms of thermoinactivation of hyperthermophilic L-asparaginase (L-ASNase) from archaea Thermococcus sibiricus (TsA) in comparison with bacterial L-ASNases from Melioribacter roseus (MrA) and Rhodospirillum rubrum (RrA). The catalytic parameters of L-asparagine hydrolysis under optimal conditions (pH 9) were determined for these enzymes by circular dichroism (CD) spectroscopy. TsA showed the highest activity among the studied L-ASNases (640 IU/mg at 90 °C). Thermo-inactivation kinetics were studied at temperatures close to the enzyme optimum: the first-order inactivation constants were 0.065 min−1 (TsA), 0.011 min−1 (MrA), and 0.026 min−1 (RrA). In contrast to RrA and MrA, aggregation was detected as one of the thermoinactivation mechanisms for TsA. From the analysis of thermograms obtained with CD spectroscopy, the melting temperatures (Tm) for RrA, MrA, and TsA were determined as 50, 69, and 89 °C, respectively. A significant increase in the percentage of β-structures for TsA during heating (from 8 to 16%) indicating aggregation was observed in the interval from 70 to 100 °C. For RrA and MrA this value did not increase. Changes in the tertiary structure of the enzymes during heating were monitored by fluorescence spectroscopy. Thermal inactivation of RrA and MrA were accompanied by changes in the tertiary structure. For TsA, the observed denaturation enthalpy (ΔH) was 346 kJ/mol, which was 1.5–2 times higher than the same values for RrA and MrA. The study of the specific thermoinactivation mechanisms and structural- features in hyperthermophilic enzymes in comparison with mesophilic ones allows us to shed light on the molecular adaptation variants of the enzyme to function at high temperatures.

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L-asparaginase from Aspergillus oryzae spp.: effects of production process and biochemical parameters

Cited by 10 publications

References 44 publications

Exploring the causal relationship between glutamine metabolism and leukemia risk: a Mendelian randomization and LC-MS/MS analysis

Exploring the causal relationship between glutamine metabolism and leukemia risk: a Mendelian randomization and LC-MS/MS analysis

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

Thermal Inactivation Mechanism and Structural Features Providing Enhanced Thermal Stability of Hyperthermophilic Thermococcus sibiricus L-Asparaginase in Comparison with Mesophilic and Thermophilic L-Asparaginases

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