Miguel Duarte Toscano

Novozymes · Protein Diversity

Application of enzymatic biofuel cells (EBFCs) in wearable or implantable biomedical devices requires flexible and biocompatible electrode materials. To this end, freestanding and low-cost graphene paper is emerging among the most promising support materials. In this work, we have exploited the potential of using graphene paper with two-dimensional...

We report on a hybrid bioelectrochemical system that integrates an energy converting part, viz. a glucose/oxygen enzymatic fuel cell, with a charge-storing component, in which the redox features of the immobilized redox protein cytochrome c (cyt c) were utilized. Bilirubin oxidase and pyrroloquinoline quinone-dependent glucose dehydrogenase (PQQ-GD...

We present a fuel-independent self-charging biosupercapacitor comprising an oxygen reducing enzymatic biocathode and an opposing bioelectrode, where the supercapacitive properties of immobilized protein were utilized. Our findings disclose a novel...

Integrating photosynthetic cell components with nanostructured materials can facilitate the conversion of solar energy into electric power for creating sustainable carbon-neutral energy sources. With the aim at exploring efficient photoinduced biocatalytic energy conversion systems, we have used an amidated carbon nanotube (aCNT) networked matrix t...

We present a transparent and flexible self-charging biosupercapacitor based on an optimised mediator- and membrane-free enzymatic glucose/oxygen biofuel cell. Indium tin oxide (ITO) nanoparticles were spray-coated on transparent conducting ITO supports resulting in a flocculent, porous and nanostructured electrode surface. By this, high capacitive...

We detail a mediator- and membrane-free enzymatic glucose/oxygen biofuel cell based on transparent and nanostructured conducting supports. Chemically modified indium tin oxide nanoparticle modified electrodes were used to substantially increase the active surface area without significantly compromising transparency. Two different procedures for sur...

The front cover artwork is provided by Prof. Lo Gorton from Lund University (Sweden) and his co-workers. The image shows mutated cellobiose dehydrogenase (CDH) immobilized on a graphite electrode and how preferentially glucose is oxidized by this enzyme. Read the full text of the Article at 10.1002/celc.201600781.

The Front Cover picture shows mutated cellobiose dehydrogenase (CDH) immobilized on a graphite electrode and how preferentially glucose is oxidized by this enzyme. The enzyme consists of a larger catalytic dehydrogenase domain (grey) containing FAD as cofactor (orange) and a smaller cytochrome domain (lavender) containing heme as cofactor (yellow)...

The interface between protein and material surface is of great research interest in applications varying from implants, tissue engineering to bioelectronics. Maintaining functionality of bioelements depends greatly on the immobilization process. In the present study direct electron transfer of cellobiose dehydrogenase from Humicola insolens (HiCDH)...

Cellobiose dehydrogenase (CDH) is a fungal extracellular flavocytochrome capable of direct electron transfer (DET). Unlike other CDHs, the pH optimum for CDHs from Corynascus thermophilus (CtCDH) and Humicola insolens (HiCDH) is close to the human physiological pH in blood (7.4). These are, therefore, interesting candidates for glucose measurements...

High surface area graphene electrodes were prepared by simultaneous electrodeposition and electroreduction of graphene oxide. The electrodeposition process was optimized in terms of pH and conductivity of the solution and the obtained graphene electrodes were characterized by X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy...

The present invention relates to isolated polypeptides having xylanase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Here for the first time, we detail self-contained (wireless and self-powered) biodevices with wireless signal transmission. Specifically, we demonstrate the operation of self-sustained carbohydrate and oxygen sensitive biodevices, consisting of a wireless electronic unit, radio transmitter and separate sensing bioelectrodes, supplied with electrica...

Direct electron transfer (DET) between cellobiose dehydrogenase from Humicola insolens ascomycete (HiCDH) and gold nanoparticles (AuNPs) was achieved by modifying AuNPs with a novel, positively charged thiol N-(6-mercapto)hexylpyridinium (MHP). The DET enabled the use of the HiCDH enzyme as an anodic biocatalyst in the design of a mediatorless carb...

Gold disk electrodes modified with gold nanoparticles have been used as a scaffold for the covalent immobilization of bilirubin oxidase. The nanostructured bioelectrodes were tested as mediator-less biosensors for oxygen in a buffer that mimics the content and the composition of human physiological fluids. Chronoamperometry measurements showed a de...

After initial testing and optimization of anode biocatalysts, a membraneless glucose/oxygen enzymatic biofuel cell possessing high coulombic efficiency and power output was fabricated and characterized. Two sugar oxidizing enzymes, namely, pyranose dehydrogenase from Agaricus meleagris (AmPDH) and flavodehydrogenase domains of various cellobiose de...

Small but faster: A small laccase from Streptomyces coelicolor (SLAC) has been engineered by structure-based design and site-directed mutagenesis to improve the activity on commercially relevant substrates. The variants generated showed up to 40-fold increased efficiency on 2,6-dimethoxyphenol and the ability to use mediators with considerably high...

A microscale membrane-less biofuel cell, capable of generating electrical energy from human lachrymal liquid, was developed by utilizing the ascorbate and oxygen naturally present in tears as fuel and oxidant. The biodevice is based on three-dimensional nanostructured gold electrodes covered with abiotic (conductive organic complex) and biological...

Low-density graphite electrodes have been covalently modified with gold nanoparticles and used as scaffold for the oriented immobilization of Myrothecium verrucaria bilirubin oxidase. The developed nanobioelectrodes have been tested as a biocathode in a human-serum surrogate buffer that mimics physiological conditions. The nanostructured bioelectro...

Threonine aldolases are versatile pyridoxal-5′-phosphate (PLP)-dependent enzymes key to glycine, serine and threonine metabolism. Because they catalyze the reversible addition of glycine to an aldehyde to give β-hydroxy-α-amino acids, they are also attractive as biotechnological catalysts for the diastereoselective synthesis of many pharmaceuticall...

Cellobiose dehydrogenase (CDH) has recently become a redox enzyme at focus in bioelectrochemistry especially for the construction of sugar biosensors and biofuel cell anodes. The present study shows that an increase in the CaCl2 concentration to up to 100 mM led to an increase in the maximal catalytic current generated by two different Ascomycete a...

Many enzymes catalyze reactions with multiple chemical steps, requiring the stabilization of multiple transition states during catalysis. Such enzymes must strike a balance between the conformational reorganization required to stabilize multiple transition states of a reaction and the confines of a preorganized active site in the polypeptide tertia...

Although nature evolves its catalysts over millions of years, enzyme engineers try to do it a bit faster. Enzyme active sites provide highly optimized microenvironments for the catalysis of biologically useful chemical transformations. Consequently, changes at these centers can have large effects on enzyme activity. The prediction and control of th...

Während die Natur ihre Katalysatoren über Millionen von Jahren entwickelt hat, versuchen Enzymingenieure, dies ein wenig schneller zu bewerkstelligen. Die aktive Tasche eines Enzyms bildet eine stark optimierte Mikroumgebung für die Katalyse chemischer Transformationen in biologischen Systemen – folglich können Änderungen in diesen Zentren die Enzy...

Chop and change: Site-directed mutagenesis has been employed to optimize the activity of an engineered pyridoxal phosphate-dependent aldolase and to invert its inherent threo selectivity in the cleavage of D-β-phenylserines. The modification of the active-site residues generates significant retroaldol activity that compares favorably with that of n...

We describe the rational design of a novel, highly potent inhibitor of type II dehydroquinase, the dicarboxylate 6. The incorporation of a carboxylate at the 3-position mimics the putative enolate intermediate in the reaction mechanism, and allows a potential electrostatic binding interaction with the arginine on the active site flap. This results...

Selective inhibitors of type II dehydroquinase were rationally designed to explore a second binding-pocket in the active-site. The molecular modelling, synthesis, inhibition studies and crystal structure determination are described.

Aromatic analogues of chorismate were synthesised as potential inhibitors of anthranilate synthase. Molecular modelling using GOLD2.1 showed that these analogues docked into the active site of Serratia marcescens anthranilate synthase in the same conformation as chorismate. Most compounds were found to be micromolar inhibitors of S. marcescens anth...

A series of 1-substituted and 4-substituted benzyl analogues of the known inhibitor (1S,3R,4R)-1,3,4-trihydroxy-5-cyclohexene-1-carboxylic acid has been synthesized and tested as inhibitors of Streptomyces coelicolor type II dehydroquinase. The solid-phase syntheses of 18 new analogues are reported. The most potent inhibitor, 2-nitrobenzyloxy analo...

Inhibitors of type II dehydroquinase were designed to straddle the two distinct binding sites identified for the inhibitor (1S,3R,4R)-1,3,4-trihydroxy-5-cyclohexene-1-carboxylic acid and a glycerol molecule in a crystallographic study of the Streptomyces coelicolor enzyme. A number of compounds were designed to incorporate characteristics of both l...