Site directed immobilization of glucose-6-phosphate dehydrogenase via thiol-disulfide interchange: Influence on catalytic activity of cysteines introduced at different positions

Simons, Jan Robert; Mosisch, Martin; Torda, Andrew E.; Hilterhaus, Lutz

doi:10.1016/j.jbiotec.2013.06.002

Cited by 25 publications

(12 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Notably, the introduction of an additional cysteine residue to the C-terminal (mutant C-c) not only enhanced the immobilized enzyme activity, which was about 2.6 fold of wtFDH, but also improved the immobilization yield dramatically. This improvement may be caused by the favorable orientation of the immobilized enzyme on PD-IONPs (Simons et al, 2013). Table 1 summarizes the comparative data on the enzymatic properties of the mutant C-c and wtFDH.…”

Section: Effect Of Site-directed Mutagenesis On Immobilized Enzyme Acmentioning

confidence: 99%

Enhancement of the activity of enzyme immobilized on polydopamine-coated iron oxide nanoparticles by rational orientation of formate dehydrogenase

Gao

Zhao

et al. 2014

Journal of Biotechnology

View full text Add to dashboard Cite

Section: Effect Of Site-directed Mutagenesis On Immobilized Enzyme Acmentioning

confidence: 99%

Enhancement of the activity of enzyme immobilized on polydopamine-coated iron oxide nanoparticles by rational orientation of formate dehydrogenase

Gao

Zhao

et al. 2014

Journal of Biotechnology

View full text Add to dashboard Cite

“…In addition, immobilization may provide other benefits such as creating compatible environment for enzyme, and avoiding multi-layer packing of enzymes etc., such that improved specific activity may be observed after immobilization in certain cases [4]. Enzyme orientation during immobilization may play a critical role in the recovered activity, for example, when the substrate is large and the active center is oriented towards the surface of the support this may raise some steric hindrances, or when the interaction of a specific area of the enzyme with the enzyme may drive to their inactivation [5]. Modifying enzymes by adding a tag can maintain enzyme activity and avoid enzyme inactivation caused by chemical reactions.…”

Section: Introductionmentioning

confidence: 99%

Site-Specific, Covalent Immobilization of Dehalogenase ST2570 Catalyzed by Formylglycine-Generating Enzymes and Its Application in Batch and Semi-Continuous Flow Reactors

et al. 2016

View full text Add to dashboard Cite

Formylglycine-generating enzymes can selectively recognize and oxidize cysteine residues within the sulfatase sub motif at the terminus of proteins to form aldehyde-bearing formylglycine (FGly) residues, and are normally used in protein labeling. In this study, an aldehyde tag was introduced to proteins using formylglycine-generating enzymes encoded by a reconstructed set of the pET28a plasmid system for enzyme immobilization. The haloacid dehalogenase ST2570 from Sulfolobus tokodaii was used as a model enzyme. The C-terminal aldehyde-tagged ST2570 (ST2570CQ) exhibited significant enzymological properties, such as new free aldehyde groups, a high level of protein expression and improved enzyme activity. SBA-15 has widely been used as an immobilization support for its large surface and excellent thermal and chemical stability. It was functionalized with amino groups by aminopropyltriethoxysilane. The C-terminal aldehyde-tagged ST2570 was immobilized to SBA-15 by covalent binding. The site-specific immobilization of ST2570 avoided the chemical denaturation that occurs in general covalent immobilization and resulted in better fastening compared to physical adsorption. The site-specific immobilized ST2570 showed 3-fold higher thermal stability, 1.2-fold higher catalytic ability and improved operational stability than free ST2570. The site-specific immobilized ST2570 retained 60% of its original activity after seven cycles of batch operation, and it was superior to the ST2570 immobilized to SBA-15 by physical adsorption, which loses 40% of its original activity when used for the second time. It is remarkable that the site-specific immobilized ST2570 still retained 100% of its original activity after 10 cycles of reuse in the semi-continuous flow reactor. Overall, these results provide support for the industrial-scale production and application of site-specific, covalently immobilized ST2570.

show abstract

“…Also, studies done by Simons et al [149] revealed that G6PD could be efficiently immobilized via genetically engineered cysteine, producing higher enzyme activity than the regular immobilized G6PD. Therefore, implementation of the improved technique of immobilized G6PD from Pyrococcus furiosus may tend to increase the yield and lower cost of constantly isolating G6PD.…”

Section: In Enzyme Immobilization For Biofuel Productionmentioning

confidence: 99%

Updates on Glucose 6-Phosphate Dehydrogenase (G6PD): From Prokaryotes to Human

2016

IJSR

View full text Add to dashboard Cite

Glucose 6-phosphate dehydrogenase (G6PD) is the rate determining enzyme that catalyzes the first step in pentose phosphate pathway (PPP) which produces NADPH that serves as reducing agent for reductive biosynthetic reactions such as steroids, fatty acids and nucleotides syntheses. Been ubiquitous, it has been found in wide range of organisms, ranging from prokaryotic organisms to even higher animals like human. Numerous G6PD isoenzymes have been isolated, purified and well characterised. However, obtained information such as its kinetic parameters and biological properties have not been judiciously appropriated for biotechnological/medical applications. This review thus concisely showcases G6PD, focusing on some sources where it has been isolated, purified and characterized, physicochemical/biochemical properties, the benefits and adverse effects its evolution may confer, and the possible biotechnological applications/prospects for the enzyme.

show abstract

Site directed immobilization of glucose-6-phosphate dehydrogenase via thiol-disulfide interchange: Influence on catalytic activity of cysteines introduced at different positions

Cited by 25 publications

References 57 publications

Enhancement of the activity of enzyme immobilized on polydopamine-coated iron oxide nanoparticles by rational orientation of formate dehydrogenase

Enhancement of the activity of enzyme immobilized on polydopamine-coated iron oxide nanoparticles by rational orientation of formate dehydrogenase

Site-Specific, Covalent Immobilization of Dehalogenase ST2570 Catalyzed by Formylglycine-Generating Enzymes and Its Application in Batch and Semi-Continuous Flow Reactors

Updates on Glucose 6-Phosphate Dehydrogenase (G6PD): From Prokaryotes to Human

Contact Info

Product

Resources

About