Self-Immobilizing Fusion Enzymes for Compartmentalized Biocatalysis

ACS Catalysis ◽  
2017 ◽  
Vol 7 (11) ◽  
pp. 7866-7872 ◽  
Author(s):  
Theo Peschke ◽  
Marc Skoupi ◽  
Teresa Burgahn ◽  
Sabrina Gallus ◽  
Ishtiaq Ahmed ◽  
...  
Keyword(s):  
Fusion Enzymes

scholarly journals Amperometric Biosensors Based on Direct Electron Transfer Enzymes

Molecules ◽  
2021 ◽  
Vol 26 (15) ◽  
pp. 4525
Author(s):  
Franziska Schachinger ◽  
Hucheng Chang ◽  
Stefan Scheiblbrandner ◽  
Roland Ludwig
Keyword(s):  
Electron Transfer ◽  
Electrode Materials ◽  
Direct Electron Transfer ◽  
Accurate Determination ◽  
Direct Electrochemistry ◽  
Product Analysis ◽  
Amperometric Biosensors ◽  
Chemical Surface ◽  
Fusion Enzymes ◽  
Cost Efficient

The accurate determination of analyte concentrations with selective, fast, and robust methods is the key for process control, product analysis, environmental compliance, and medical applications. Enzyme-based biosensors meet these requirements to a high degree and can be operated with simple, cost efficient, and easy to use devices. This review focuses on enzymes capable of direct electron transfer (DET) to electrodes and also the electrode materials which can enable or enhance the DET type bioelectrocatalysis. It presents amperometric biosensors for the quantification of important medical, technical, and environmental analytes and it carves out the requirements for enzymes and electrode materials in DET-based third generation biosensors. This review critically surveys enzymes and biosensors for which DET has been reported. Single- or multi-cofactor enzymes featuring copper centers, hemes, FAD, FMN, or PQQ as prosthetic groups as well as fusion enzymes are presented. Nanomaterials, nanostructured electrodes, chemical surface modifications, and protein immobilization strategies are reviewed for their ability to support direct electrochemistry of enzymes. The combination of both biosensor elements—enzymes and electrodes—is evaluated by comparison of substrate specificity, current density, sensitivity, and the range of detection.

scholarly journals Aromatic Halogenation by Using Bifunctional Flavin Reductase-Halogenase Fusion Enzymes

ChemBioChem ◽  
2017 ◽  
Vol 18 (21) ◽  
pp. 2099-2103 ◽  
Author(s):  
Mary C. Andorfer ◽  
Ketaki D. Belsare ◽  
Anna M. Girlich ◽  
Jared C. Lewis
Keyword(s):  
Flavin Reductase ◽  
Fusion Enzymes

In-fusion expression and characterization of β-xylanase and β-1,3-1,4-glucanase in Pichia pastoris

Biologia ◽  
2012 ◽  
Vol 67 (4) ◽  
Author(s):  
Jiayun Qiao ◽  
Yunhe Cao
Keyword(s):  
Bacillus Subtilis ◽  
Pichia Pastoris ◽  
Fusion Protein ◽  
Fusion Expression ◽  
Chimeric Genes ◽  
Fusion Enzyme ◽  
Fusion Enzymes ◽  
Aspergillus Sulphureus

AbstractTwo chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Kn higher kcat. Despite some variations, common optima were generally 50°C and pH 3.4 for the XynA moiety and parent, and 40°C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.

Degradation of cellulose using two fusion enzymes generated by glucan-polymerization enzyme

2016 ◽  
Vol 33 ◽  
pp. S103
Author(s):  
Cheon-Seok Park ◽  
Ye Ji Lee ◽  
Dong Hyun Jung ◽  
Jong Hyun Jung ◽  
Dong Ho Seo
Keyword(s):  
Fusion Enzymes

scholarly journals Characterization of hybrid proteins consisting of the catalytic domains of Clostridium and Ruminococcus endoglucanases, fused to Pseudomonas non-catalytic cellulose-binding domains

1991 ◽  
Vol 279 (3) ◽  
pp. 787-792 ◽  
Author(s):  
D M Poole ◽  
A J Durrant ◽  
G P Hazlewood ◽  
H J Gilbert
Keyword(s):  
Catalytic Domain ◽  
Binding Capacity ◽  
Binding Domains ◽  
Hybrid Proteins ◽  
C Terminus ◽  
N Terminus ◽  
Fusion Enzymes ◽  
Cellulose Binding

The N-terminal 160 or 267 residues of xylanase A from Pseudomonas fluorescens subsp. cellulosa, containing a non-catalytic cellulose-binding domain (CBD), were fused to the N-terminus of the catalytic domain of endoglucanase E (EGE') from Clostridium thermocellum. A further hybrid enzyme was constructed consisting of the 347 N-terminal residues of xylanase C (XYLC) from P. fluorescens subsp. cellulosa, which also constitutes a CBD, fused to the N-terminus of endoglucanase A (EGA) from Ruminococcus albus. The three hybrid enzymes bound to insoluble cellulose, and could be eluted such that cellulose-binding capacity and catalytic activity were retained. The catalytic properties of the fusion enzymes were similar to EGE' and EGA respectively. Residues 37-347 and 34-347 of XYLC were fused to the C-terminus of EGE' and the 10 amino acids encoded by the multiple cloning sequence of pMTL22p respectively. The two hybrid proteins did not bind cellulose, although residues 39-139 of XYLC were shown previously to constitute a functional CBD. The putative role of the P. fluorescens subsp. cellulosa CBD in cellulase action is discussed.

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