Intracellular-to-extracellular localization switch of acidic lipase in <i>Enterobacter cloacae</i>: evaluation of production kinetics and enantioselective esterification potential for pharmaceutical applications

Asitok, Atim; Ekpenyong, Maurice; Ogarekpe, Nkpa; Antigha, Richard; Takon, Iquo; Rao, Anil V.; Iheanacho, Juliet; Antai, S. P.

doi:10.1080/10826068.2022.2114010

Cited by 4 publications

(2 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Stability of Fusant-06 l -ASNase to organic solvents strongly and significantly correlated with the octanol–water partition coefficients, log P (Sangster 1989 ) of the solvents (Additional file 1 : Figure S5), suggesting that enzyme stability reduced with increase in solvent hydrophobicity (Asitok et al 2022b ).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

et al. 2023

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The sequence of amino acids in l -ASNase from Fusant-06 was determined by Agilent amino acid analyzer (Asitok et al 2022b ). Sequences obtained were compared with NCBI sequences for Aspergillus l -ASNases and with those of the wild Aspergillus strains.…”

Section: Methodsmentioning

confidence: 99%