scholarly journals Engineered catalytic biofilms: Site-specific enzyme immobilization ontoE. colicurli nanofibers

2015 ◽  
Vol 112 (10) ◽  
pp. 2016-2024 ◽  
Author(s):  
Zsofia Botyanszki ◽  
Pei Kun R. Tay ◽  
Peter Q. Nguyen ◽  
Martin G. Nussbaumer ◽  
Neel S. Joshi
Keyword(s):  
Enzyme Immobilization ◽  
Specific Enzyme ◽  
Site Specific

Design of a Specific Peptide Tag that Affords Covalent and Site-Specific Enzyme Immobilization Catalyzed by Microbial Transglutaminase

2005 ◽  
Vol 6 (4) ◽  
pp. 2299-2304 ◽  
Author(s):  
Jo Tominaga ◽  
Noriho Kamiya ◽  
Satoshi Doi ◽  
Hirofumi Ichinose ◽  
Tatsuo Maruyama ◽  
...  
Keyword(s):  
Enzyme Immobilization ◽  
Microbial Transglutaminase ◽  
Specific Enzyme ◽  
Site Specific ◽  
Specific Peptide ◽  
Peptide Tag

Surface functionalized halloysite nanotubes decorated with silver nanoparticles for enzyme immobilization and biosensing

2016 ◽  
Vol 4 (15) ◽  
pp. 2553-2560 ◽  
Author(s):  
Siva Kumar-Krishnan ◽  
A. Hernandez-Rangel ◽  
Umapada Pal ◽  
O. Ceballos-Sanchez ◽  
F. J. Flores-Ruiz ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Enzyme Immobilization ◽  
Halloysite Nanotubes ◽  
Covalent Immobilization ◽  
Site Specific

Illustrating the selective modification of the HNTs for the incorporation of AgNPs and site-specific covalent immobilization of the GOx enzymes.

scholarly journals Molecular Decoration of Ceramic Supports for Highly Effective Enzyme Immobilization—Material Approach

Materials ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 201
Author(s):  
Joanna Kujawa ◽  
Marta Głodek ◽  
Izabela Koter ◽  
Borys Ośmiałowski ◽  
Katarzyna Knozowska ◽  
...  
Keyword(s):  
Enzyme Immobilization ◽  
Immobilized Lipase ◽  
Material Surface ◽  
Specific Enzyme ◽  
Enzyme Loading ◽  
Enantiomeric Ratio ◽  
Highly Effective ◽  
Functionalized Materials ◽  
Specific Material ◽  
The Impact

A highly effective method was developed to functionalize ceramic supports (Al2O3 powders and membranes) using newly synthesized spacer molecules. The functionalized materials were subsequently utilized for Candida antarctica lipase B enzyme immobilization. The objective is to systematically evaluate the impact of various spacer molecules grafted onto the alumina materials will affect both the immobilization of the enzymes and specific material surface properties, critical to enzymatic reactors performance. The enzyme loading was significantly improved for the supports modified with shorter spacer molecules, which possessed higher grafting effectiveness on the order of 90%. The specific enzyme activity was found to be much higher for samples functionalized with longer modifiers yielding excellent enantioselectivity >97%. However, the enantiomeric ratio of the immobilized lipase was slightly lower in the case of shorter spacer molecules.

scholarly journals Design and Construction of an Effective Expression System with Aldehyde Tag for Site-Specific Enzyme Immobilization

Catalysts ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 410
Author(s):  
Fang Wang ◽  
Rong Li ◽  
Hui Jian ◽  
Zihao Huang ◽  
Yingwu Wang ◽  
...  
Keyword(s):  
Immobilized Enzyme ◽  
Expression System ◽  
Aldehyde Group ◽  
Covalent Immobilization ◽  
Specific Enzyme ◽  
Site Specific ◽  
Knock Out ◽  
Sulfolobus Tokodaii ◽  
Immobilization Technology ◽  
Lipase A

In recent years, the development and application of site-specific immobilization technology for proteins have undergone significant advances, which avoids the unwanted and random covalent linkage between the support and active site of protein in the covalent immobilization. Formylglycine generating enzyme (FGE) can transform the cysteine from a conversed 6-amino-acid sequence CXPXR into formylglycine with an aldehyde group (also termed as “aldehyde tag”). Based on the frame of pET-28a, the His-tags were replaced with aldehyde tags. Afterward, a set of plasmids were constructed for site-specific covalent immobilization, their His-tags were knock out (DH), or were replaced at different positions: N-terminal (NQ), C-terminal (CQ), or both (DQ) respectively. Three different enzymes, thermophilic acyl aminopeptidase (EC 3.4.19.1) from Sulfolobus tokodaii (ST0779), thermophilic dehalogenase (EC 3.8.1.2) from Sulfolobus tokodaii (ST2570), and Lipase A (EC 3.1.1.3) from Bacillus subtilis (BsLA) were chosen as model enzymes to connect with these plasmid systems. The results showed that different aldehyde-tagged enzymes can be successfully covalently attached to different carriers modified with an amino group, proving the universality of the method. The new immobilized enzyme also presented better thermostability and reutilization than those of the free enzyme.

Specific Enzyme Immobilization Approaches and Their Application with Nanomaterials

2012 ◽  
Vol 55 (16-18) ◽  
pp. 1146-1156 ◽  
Author(s):  
Wenshan Liu ◽  
Liang Wang ◽  
Rongrong Jiang
Keyword(s):  
Enzyme Immobilization ◽  
Specific Enzyme

Large-cluster and combined fluorescent and gold immunoprobes

1996 ◽  
Vol 54 ◽  
pp. 892-893
Author(s):  
Richard D. Powell ◽  
James F. Hainfeld ◽  
Carol M. R. Halsey ◽  
David L. Spector ◽  
Shelley Kaurin ◽  
...  
Keyword(s):  
Metal Cluster ◽  
Sulfonyl Chloride ◽  
Gel Filtration ◽  
Cross Linking ◽  
Site Specific ◽  
Fab Fragments ◽  
Cluster Label ◽  
Covalently Linked ◽  
Texas Red ◽  
Fold Excess

Two new types of covalently linked, site-specific immunoprobes have been prepared using metal cluster labels, and used to stain components of cells. Combined fluorescein and 1.4 nm “Nanogold” labels were prepared by using the fluorescein-conjugated tris (aryl) phosphine ligand and the amino-substituted ligand in the synthesis of the Nanogold cluster. This cluster label was activated by reaction with a 60-fold excess of (sulfo-Succinimidyl-4-N-maleiniido-cyclohexane-l-carboxylate (sulfo-SMCC) at pH 7.5, separated from excess cross-linking reagent by gel filtration, and mixed in ten-fold excess with Goat Fab’ fragments against mouse IgG (obtained by reduction of F(ab’)2 fragments with 50 mM mercaptoethylamine hydrochloride). Labeled Fab’ fragments were isolated by gel filtration HPLC (Superose-12, Pharmacia). A combined Nanogold and Texas Red label was also prepared, using a Nanogold cluster derivatized with both and its protected analog: the cluster was reacted with an eight-fold excess of Texas Red sulfonyl chloride at pH 9.0, separated from excess Texas Red by gel filtration, then deprotected with HC1 in methanol to yield the amino-substituted label.

The challenge of detecting modifications on proteins

2020 ◽  
Vol 64 (1) ◽  
pp. 135-153 ◽  
Author(s):  
Lauren Elizabeth Smith ◽  
Adelina Rogowska-Wrzesinska
Keyword(s):  
Mass Spectrometry ◽  
Protein Function ◽  
Chemical Properties ◽  
Lysine Acetylation ◽  
Mass Determination ◽  
Post Translational Modifications ◽  
Site Specific ◽  
The Common

Abstract Post-translational modifications (PTMs) are integral to the regulation of protein function, characterising their role in this process is vital to understanding how cells work in both healthy and diseased states. Mass spectrometry (MS) facilitates the mass determination and sequencing of peptides, and thereby also the detection of site-specific PTMs. However, numerous challenges in this field continue to persist. The diverse chemical properties, low abundance, labile nature and instability of many PTMs, in combination with the more practical issues of compatibility with MS and bioinformatics challenges, contribute to the arduous nature of their analysis. In this review, we present an overview of the established MS-based approaches for analysing PTMs and the common complications associated with their investigation, including examples of specific challenges focusing on phosphorylation, lysine acetylation and redox modifications.

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