Immobilization of cyclodextrin glucanotransferase on aminopropyl-functionalized silica-coated superparamagnetic nanoparticles

Ibrahim, Abdelnasser S.S.; Al‐Salamah, Ali A.; El‐Toni, Ahmed Mohamed; El-Tayeb, Mohamed A.; Elbadawi, Yahya B.

doi:10.2225/vol16-issue6-fulltext-8

Cited by 16 publications

(12 citation statements)

References 48 publications

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“…NPST-AK15 protease immobilization by physical adsorption was carried using HCMSS-non, HCMSS–C 2 H 5 and HCMSS–NH 2 according to Ibrahim et al [ 37 ]. Briefly, 50 mg of carriers were dispersed in 1 mL of glycine buffer (50 mM, pH 8) that contained the purified NPST-AK15 protease and the suspensions were kept for overnight at 4 °C with gentle shaking.…”

Section: Methodsmentioning

confidence: 99%

Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres

Ibrahim

Al‐Salamah

El‐Toni

et al. 2016

IJMS

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The stability and reusability of soluble enzymes are of major concerns, which limit their industrial applications. Herein, alkaline protease from Bacillus sp. NPST-AK15 was immobilized onto hollow core-mesoporous shell silica (HCMSS) nanospheres. Subsequently, the properties of immobilized proteases were evaluated. Non-, ethane- and amino-functionalized HCMSS nanospheres were synthesized and characterized. NPST-AK15 was immobilized onto the synthesized nano-supports by physical and covalent immobilization approaches. However, protease immobilization by covalent attachment onto the activated HCMSS–NH2 nanospheres showed highest immobilization yield (75.6%) and loading capacity (88.1 μg protein/mg carrier) and was applied in the further studies. In comparison to free enzyme, the covalently immobilized protease exhibited a slight shift in the optimal pH from 10.5 to 11.0, respectively. The optimum temperature for catalytic activity of both free and immobilized enzyme was seen at 60 °C. However, while the free enzyme was completely inactivated when treated at 60 °C for 1 h the immobilized enzyme still retained 63.6% of its initial activity. The immobilized protease showed higher Vmax, kcat and kcat/Km, than soluble enzyme by 1.6-, 1.6- and 2.4-fold, respectively. In addition, the immobilized protease affinity to the substrate increased by about 1.5-fold. Furthermore, the enzyme stability in various organic solvents was significantly enhanced upon immobilization. Interestingly, the immobilized enzyme exhibited much higher stability in several commercial detergents including OMO, Tide, Ariel, Bonux and Xra by up to 5.2-fold. Finally, the immobilized protease maintained significant catalytic efficiency for twelve consecutive reaction cycles. These results suggest the effectiveness of the developed nanobiocatalyst as a candidate for detergent formulation and peptide synthesis in non-aqueous media.

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Section: Methodsmentioning

confidence: 99%

Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres

Ibrahim

Al‐Salamah

El‐Toni

et al. 2016

IJMS

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“…It is characterized by uniform pore system, tunable and easy to be functionalized, biocompatibility and low cytotoxicity (Zhou & Hartmann, 2013). In addition, magnetic silica can be prepared by silica coating on magnetic nanoparticle cores and hence the immobilized enzyme can be easily separated from the reaction solution by application of an external magnetic field (Ibrahim et al, 2014 andKumari &.My group and I have explored utilization of several silica based nanoparticles for immobilization of different enzymes such as immobilization of cyclodextrin glucanotransferase (CGTase) onto aminopropyl functionalized silica coated superparamagnetic nanoparticles (Ibrahim et al, 2013) and functionalized magnetic double mesoporous core-shell silica nanospheres (Ibrahim et al, 2014). The immobilized CGTase exhibited significant improvement of enzyme kinetics and stability parameters, high operation stability and can be recovered easily from the bulk solution using magnetic field (Fig.…”

Section: Nanostructured Materials and Nanobiocatalysismentioning

confidence: 99%

“…TEM images of (a) magnetic magnetite (Fe 3 O 4 ); (b) NH 2 -SiO 2 coated magnetic Fe 3 O 4 nanoparticles. (C) Separation of CGTase immobilized on aminopropyl functionalized silica-coated superparamagnetic nanoparticles by an external magnetic field(Ibrahim et al, 2013; The Authors group).…”

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confidence: 99%

Recent Trends for Discovery and Enhancement of Enzyme Function: A Review

Ibrahim

El‐Dewany

2018

Egypt. J. Microbiol.

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R ECENTLY, there is a growing recognition of enzymes utilization as ecofriendly alternative for chemical catalysts in various industrial sectors. Two main approaches are used to expand the enzymes utilization in various industrial applications including (i) Searching nature for discovery of better performing novel biocatalysts and (ii) Improvement of the enzyme function and properties. The recent advances in biocatalyst discovery and engineering have led to an increase of the efficiency and functional diversityof enzymes. Therefore, enzyme application is expanding to broader industrial processes, including those which were previously restricted to the classical chemical catalysts. Furthermore, there is a significant success in developing made-to-order enzymes that meet the industrial process conditions requirements, leading to more efficient and cost-effective processes. Here I highlight state-of-the-art strategies and tools for novel enzyme discovery and function enhancement, including enzyme engineering at the molecular level through different approaches such as directed evolution, rational design, semi rational design and computational de novo enzyme design; in addition to the recently developed field, nanobiocatalysis.

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“…Then, the solvent was evaporated to obtain the AFB1-oxime active ester. [32] 2.3 | Preparation of magnetic nanoparticles Magnetic nanoparticles (MNPs) were prepared by solvothermal synthesis reaction [33] with some modifications. Briefly, FeCl 3 ·6H 2 O (1.35 g) was dissolved in ethylene glycol (40 mL) to form a clear solution, followed by addition of polyethylene glycol (1.0 g) and sodium acetate (3.6 g) as a stabilizing agent.…”

Section: Synthesis Of Afb1 Haptenmentioning

confidence: 99%

The chemiluminescence immunoassay for aflatoxin B1 based on functionalized magnetic nanoparticles with two strategies of antigen probe immobilization

2016

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A rapid and sensitive chemiluminescence immunoassay (CLIA) based on magnetic nanoparticles (MNPs) was developed to detect aflatoxin B1 (AFB1), which is a potent carcinogen in nature. We prepared monodisperse MNPs (300 nm in diameter) according to the solvothermal synthesis reaction and the MNPs were coated with silica by the Stöber method. Triethox was used as a one-step carboxylation reagent, and 3-aminopropyltriethoxysilane (APTES) an amination reagent, to modify the MNPs. We prepared two types of solid phase antigens using the above synthesized functionalized MNPs coupled with the later prepared AFB1-oxime active ester and the purchased BSA-AFB1 respectively. 2',6'-dimethylcarbonylphenyl-10-sulfopropylacridinium-9-carboxylate 4'-N-hydroxysuccinimide (4'-NHS) ester (NSP-DMAE-NHS), as a novel luminescent reagent, was used to label anti-AFB1 antibodies. The two CLIA calibration curves based on the two types of solid phase antigens were obtained and compared. The acquired limit of detection (LOD) was about 0.001 ng/mL for the two functionalized MNPs-based immunoassays, and the half maximal inhibitory concentration (IC ) was 0.51 ng/mL for the MNPs-AFB1-based method and 0.72 ng/mL for the MNPs-BSA-AFB1-based method.

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Immobilization of cyclodextrin glucanotransferase on aminopropyl-functionalized silica-coated superparamagnetic nanoparticles

Cited by 16 publications

References 48 publications

Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres

Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres

Recent Trends for Discovery and Enhancement of Enzyme Function: A Review

The chemiluminescence immunoassay for aflatoxin B1 based on functionalized magnetic nanoparticles with two strategies of antigen probe immobilization

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