Purification of bacterial inulinase in aqueous two‐phase systems

Temkov, Mishela; Strinska, Hristina; Dobrev, Georgi; Velickova, Elena; Mureșan, Vlad; Krastanov, Albert

doi:10.1002/elsc.201700174

Cited by 9 publications

(3 citation statements)

References 45 publications

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“…The normalized activity and stability of free and immobilized enzyme calculated for the same amount of enzyme has been evaluated under the same conditions in a wide range of pH (3.0-10.7); the results are shown in the Figure 5A and B. As reported in our previous study [46], the pH profile of the partially purified PEG phase free enzyme exhibits two peaks of optimal pH at 8 and 5.8, suggesting that the inulinase is present in two multiple forms. pH profile for the partially purified immobilized inulinase on the f-MWCNT prepared in this study also showed two pH optimums at 3.6 and 10 with the corresponding activity of 3.48 and 4.14 U/mg, respectively ( Figure 5A).…”

Section: Effect Of Ph On Immobilized Inulinase Activity and Stabilitymentioning

confidence: 71%

“…Inulinase was produced by submerged fermentation of Bacillus sp. 11/3, as described in Temkov et al [46]. The enzyme was partially purified by ATPS and then immobilized onto MWCNTs.…”

Section: Practical Applicationmentioning

confidence: 99%

“…11/3, as described in Temkov et al. . The enzyme was partially purified by ATPS and then immobilized onto MWCNTs.…”

Section: Introductionmentioning

confidence: 99%

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Inulinase immobilization on polyethylene glycol/polypyrrole multiwall carbon nanotubes producing a catalyst with enhanced thermal and operational stability

Temkov

Petrovski

Gjorgieva

et al. 2019

Engineering in Life Sciences

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This paper describes the development of a simple method for mixed non‐covalent and covalent bonding of partially purified inulinase on functionalized multiwall carbon nanotubes (f‐MWCNTs) with polypyrrole (PPy). The pyrrole (Py) was electrochemically polymerized on MWCNTs in order to fabricate MWCNTs/PPy nanocomposite. Two multiple forms of enzyme were bound to N‐H functional groups from PPy and ‐COO− from activated MWCNTs to yield a stable MWCNTs/PPy/PEG immobilized preparation with increased thermal stability. Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) were used to confirm functionalization of nanoparticles and immobilization of the enzyme. The immobilization yield of 85% and optimal enzyme load of 345 μg protein onto MWCNTs was obtained. The optimum reaction conditions and kinetic parameters were established using the UV‐Vis analytical assay. The best functional performance for prepared heterogeneous catalyst has been observed at pH 3.6 and 10, and at the temperatures of 60 and 80ºC. The half‐life (t1/2) of the immobilized inulinase at 60 and 80ºC was found to be 231 and 99 min, respectively. The reusability of the immobilized formulation was evaluated based on a method in which the enzyme retained 50% of its initial activity, which occurred after the eighteenth operation cycle.

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Section: Effect Of Ph On Immobilized Inulinase Activity and Stabilitymentioning

confidence: 71%

“…Inulinase was produced by submerged fermentation of Bacillus sp. 11/3, as described in Temkov et al [46]. The enzyme was partially purified by ATPS and then immobilized onto MWCNTs.…”

Section: Practical Applicationmentioning

confidence: 99%

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Inulinase immobilization on polyethylene glycol/polypyrrole multiwall carbon nanotubes producing a catalyst with enhanced thermal and operational stability

Temkov

Petrovski

Gjorgieva

et al. 2019

Engineering in Life Sciences

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Bio-utilization of agricultural residue banana plant shoot through solid state fermentation for production of inulinase using newly isolated Nothophoma anigozanthi JAM

Tippanavar

Bhat

Rebello

et al. 2023

Biomass Conv. Bioref.

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The present research was carried out with the objectives of isolating fungal strains capable of exhibiting inulinase activity from a novel source, then optimizing solid state fermentation for the inulinase production and characterizing crude enzyme. Production of inulinase is first reported in this study with a novel substrate, banana plant shoot powder, using newly isolated fungal strain Nothophoma anigozanthi JAM through solid state fermentation. The fungal strain was isolated from the root soil of a white Jamun (Syzygium cumini) tree. Optimization of the solid state fermentation was carried out by central composite design for determining the interaction and impact of the influential process on inulinase production. A mathematical model was developed based on the statistical analysis and recommended optimal process conditions of 2 g of substrate, 5 mL of moistening media, 5 mL of inoculum volume, and 96 h of fermentation time for enhanced production (2.578 U/gds). Crude inulinase enzyme exhibited an 8.0-fold increase in activity after optimization, in comparison with the activity under unoptimized conditions. The optimum pH (5.0), temperature (60 °C) for the enzyme activity, and Km (0.205 mM) and Vmax (0.333 µM/min) were determined based on the biochemical and inulin hydrolytic properties of the crude inulinase enzyme. The production of inulinase was further validated by using TLC, osazone, and Seliwanoff tests. Hence, the inulinase enzyme produced by Nothophoma anigozanthi JAM can be successfully utilized for the production of edible sugar fructose from non-edible polysaccharide inulin, present in agricultural residues such as banana shoot by solid state fermentation.

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Incorporation of electric fields to ionic liquids-based aqueous biphasic system for enhanced recovery of extracellular Kytococcus sedentarius TWHKC01 keratinase

Kee

Yim

Kondo

et al. 2021

Journal of the Taiwan Institute of Chemical Engineers

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Purification of bacterial inulinase in aqueous two‐phase systems

Cited by 9 publications

References 45 publications

Inulinase immobilization on polyethylene glycol/polypyrrole multiwall carbon nanotubes producing a catalyst with enhanced thermal and operational stability

Inulinase immobilization on polyethylene glycol/polypyrrole multiwall carbon nanotubes producing a catalyst with enhanced thermal and operational stability

Bio-utilization of agricultural residue banana plant shoot through solid state fermentation for production of inulinase using newly isolated Nothophoma anigozanthi JAM

Incorporation of electric fields to ionic liquids-based aqueous biphasic system for enhanced recovery of extracellular Kytococcus sedentarius TWHKC01 keratinase

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