Selective Immobilization of His‐Tagged Phosphomannose Isomerase on Ni Chelated Nanoparticles with Good Reusability and Activity

Dong, Weifu; Wang, Kangjing; Zhao, Laiping; Li, Ting; Wang, Qian; Ding, Zhongyang

doi:10.1002/cbic.202100497

Cited by 4 publications

(7 citation statements)

References 34 publications

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“…His-tags are most commonly used due to their ease of expression and cost effectiveness in Escherichia coli host bacteria. 19 Transition metal ions (Cu 2+ , Zn 2+ , Ni 2+ , and Co 2+ ) can form coordination bonds with imidazole residues on His, which enables their firm adsorption onto the carrier. 20 Moreover, His-tags have minimal effects on protein solubility, physical properties, or structure.…”

Section: Introductionmentioning

confidence: 99%

One-Step Purification and Immobilization of Glycosyltransferase with Zn–Ni MOF for the Synthesis of Rare Ginsenoside Rh2

Li,

Liu,

et al. 2024

ACS Appl. Mater. Interfaces

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Uridine diphosphate (UDP)-glucosyltransferases (UGTs) have received increasing attention in the field of ginsenoside Rh2 conversion. By harnessing the metal chelation between transition metal ions and imidazole groups present on Histagged enzymes, a specific immobilization of the enzyme within metal−organic frameworks (MOFs) is achieved. This innovative approach not only enhances the stability and reusability of the enzyme but also enables one-step purification and immobilization. Consequently, the need for purifying crude enzyme solutions is effectively circumvented, resulting in significant cost savings during experimentation. The use of immobilized enzymes in catalytic reactions has shown great potential for achieving higher conversion rates of ginsenoside Rh2. In this study, highly stable mesoporous Zn−Ni MOF materials were synthesized at 150 °C by a solvothermal method. The UGT immobilized on the Zn−Ni MOF (referred to as UGT@Zn−Ni MOF) exhibited superior pH adaptability and thermal stability, retaining approximately 76% of its initial activity even after undergoing 7 cycles. Furthermore, the relative activity of the immobilized enzyme remained at an impressive 80.22% even after 45 days of storage. The strong specific adsorption property of Zn−Ni MOF on His-tagged UGT was confirmed through analysis using polyacrylamide gel electrophoresis. UGT@Zn−Ni MOF was used to catalyze the conversion reaction, and the concentration of rare ginsenoside Rh2 was generated at 3.15 μg/mL. The results showed that Zn−Ni MOF is a material that can efficiently purify and immobilize His-tagged enzyme in one step and has great potential for industrial applications in enzyme purification and ginsenoside synthesis.

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Section: Introductionmentioning

confidence: 99%

One-Step Purification and Immobilization of Glycosyltransferase with Zn–Ni MOF for the Synthesis of Rare Ginsenoside Rh2

Li,

Liu,

et al. 2024

ACS Appl. Mater. Interfaces

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“…Affinity tags can be introduced by gene recombination technology, usually by fusing the “tag” into the N- or C- of the target enzymes. Among the numerous tagged recombinant proteins, more than 60% are produced by proteins including a hexahistidine tag (His-tagged) [ 27 ].…”

Section: Introductionmentioning

confidence: 99%

“…Based on previous work [ 27 ], we selected four macroporous ion exchange resins and chelated transition metal ions on the resin by static adsorption. Transition metal ions can easily form combinations with the imidazole residue of His tags.…”

Section: Introductionmentioning

confidence: 99%

Selective Immobilization of His-Tagged Enzyme on Ni-Chelated Ion Exchange Resin and Its Application in Protein Purification

Wang

Zhao

et al. 2023

IJMS

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Ion exchange resins are suitable as carriers for immobilized enzymes because of their stable physicochemical properties, appropriate particle size and pore structure, and lower loss in continuous operation. In this paper, we report the application of the Ni-chelated ion exchange resin in the immobilization of His-tagged enzyme and protein purification. Acrylic weak acid cation exchange resin (D113H) was selected from four cationic macroporous resins that could chelate the transition metal ion Ni. The maximum adsorption capacity of Ni was ~198 mg/g. Phosphomannose isomerase (PMI) can be successfully immobilized on Ni-chelated D113H from crude enzyme solution through chelation of transition metal ions with the His-tag on the enzyme. The maximum amount of immobilized PMI on the resin was ~143 mg/g. Notably, the immobilized enzyme showed excellent reusability and maintained 92% of its initial activity with 10 cycles of catalytic reaction. In addition, PMI was successfully purified using an affinity chromatography column prepared by Ni-chelated D113H, which showed the potential for the immobilization and purification process to be realized in one step.

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“…10,11 Specific requirements, such as the cost of the materials, harmlessness to both health and the environment, resistance to chemical and microbial decomposition, effectiveness of the immobilization process, effect on the catalytic efficiency, and facilitation of the operation process, should be considered to design an effective immobilization system. 12,13 The materials that have been successfully employed as supports include carbon nanotubes (CNTs), 14 metal−organic frameworks (MOFs), 15 synthetic and natural polymers, 11,16,17 hydrogels, 18 zeolites, 19 other inorganic particles, 20 etc. Among them, cellulose, 21 as a natural hydrophilic material, is one of the most promising supports for enzyme immobilization.…”

Section: ■ Introductionmentioning

confidence: 99%

“…The materials that have been successfully employed as supports include carbon nanotubes (CNTs), metal–organic frameworks (MOFs), synthetic and natural polymers, ,, hydrogels, zeolites, other inorganic particles, etc. Among them, cellulose, as a natural hydrophilic material, is one of the most promising supports for enzyme immobilization.…”

Section: Introductionmentioning

confidence: 99%

Enzyme Immobilization on a Delignified Bamboo Scaffold as a Green Hierarchical Bioreactor

Ren

Zhang

Sun

et al. 2022

ACS Sustainable Chem. Eng.

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Enzymes immobilized on cellulose-based supports provide important applications because they are convenient, economical, and stable. In this paper, we report the application of delignified bamboo as a novel cellulose-based scaffold for enzyme immobilization and bamboo enzyme-based flow bioreactors, which can drastically improve the catalytic reaction and subsequent separation. The delignified bamboo provides a superior biobenign and hierarchically structured platform that can be conveniently derivatized to conjugate functional proteins with high loading capacity. With an optimized delignification process and a modified Schiff base-dependent conjugation method, we identified an efficient approach to immobilize enzymes on bamboo. In our system, generally ∼8 mg/m 2 •g −1 proteins can be immobilized on the delignified bamboo, which is significantly higher than that of the reported methods. The immobilized enzymes on bamboo showed sustained activity under ambient conditions, excellent reusability with 13 cycles, and high stability of more than 7 weeks of storage at 4 °C, which can be readily adapted in flow reactors for both single transformation and tandem catalysis. As an example, drugs of abuse in synthetic urine samples were analyzed in β-glucuronidase (BGU)-functionalized flow reactors in 0.1 M phosphate buffer, pH 7.4, at room temperature, the condition suitable for the bioanalysis. Our work demonstrates that the delignified bamboo can serve as a readily available support for enzyme immobilization and hierarchical bioreactor construction, which will potentially increase the robustness of enzymes in analytical and synthetical applications.

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Selective Immobilization of His‐Tagged Phosphomannose Isomerase on Ni Chelated Nanoparticles with Good Reusability and Activity

Cited by 4 publications

References 34 publications

One-Step Purification and Immobilization of Glycosyltransferase with Zn–Ni MOF for the Synthesis of Rare Ginsenoside Rh2

One-Step Purification and Immobilization of Glycosyltransferase with Zn–Ni MOF for the Synthesis of Rare Ginsenoside Rh2

Selective Immobilization of His-Tagged Enzyme on Ni-Chelated Ion Exchange Resin and Its Application in Protein Purification

Enzyme Immobilization on a Delignified Bamboo Scaffold as a Green Hierarchical Bioreactor

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