English Journal

Biomolecular AND logic gate based on immobilized enzymes with precise spatial separation controlled by scanning electrochemical microscopy.

Gdor E1, Katz E, Mandler D.Author information 1Institute of Chemistry, The Hebrew University of Jerusalem , Jerusalem 91904, Israel.AbstractA surface-localized enzymatic AND gate based on scanning electrochemical microscopy was designed and studied. The gate is composed of an insulating glass surface modified with the enzyme glucose oxidase (GOx) and another surface opposing it made of a microelectrode. The latter was modified with a second enzyme, invertase (INV). The distance separating the modified microelectrode and surface controlled the output of the AND gate produced upon the biocatalytic reaction of the confined enzymes. Specifically, as the GOx-modified glass substrate entered the diffusion layer of the microelectrode, it catalyzed the regeneration of an electron-transfer mediator, ferroceniummethanol, generated electrochemically at the tip by oxidizing glucose, also generated at the tip, by catalytic cleaving of sucrose by INV. To enhance the activity of the GOx, mutarotase was added to convert α- to β-glucose to be further consumed by GOx. Hence, an increase of the current at the microelectrode was obtained by approaching the glass surface only in the presence of all the components. This is the first micrometer-sized biomolecular logic gate, of which we are aware, that is surface-confined and shows the promise held by the localization of biomolecular information-processing species.
The journal of physical chemistry. B.J Phys Chem B.2013 Dec 19;117(50):16058-65. doi: 10.1021/jp4095672. Epub 2013 Dec 9.
A surface-localized enzymatic AND gate based on scanning electrochemical microscopy was designed and studied. The gate is composed of an insulating glass surface modified with the enzyme glucose oxidase (GOx) and another surface opposing it made of a microelectrode. The latter was modified with a se
PMID 24261793

Genomewide analysis of polysaccharides degrading enzymes in 11 white- and brown-rot Polyporales provides insight into mechanisms of wood decay.

Hori C1, Gaskell J, Igarashi K, Samejima M, Hibbett D, Henrissat B, Cullen D.Author information 1Department of Biomaterials Sciences, Graduate School of Agricultural and Life Sciences, University of Tokyo, l-l-l, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan, and Institute for Microbial and Biochemical Technology, Forest Products Laboratory, 1 Gifford Pinchot Drive, Madison, Wisconsin 53726.AbstractTo degrade the polysaccharides, wood-decay fungi secrete a variety of glycoside hydrolases (GHs) and carbohydrate esterases (CEs) classified into various sequence-based families of carbohydrate-active enzymes (CAZys) and their appended carbohydrate-binding modules (CBM). Oxidative enzymes, such as cellobiose dehydrogenase (CDH) and lytic polysaccharide monooxygenase (LPMO, formerly GH61), also have been implicated in cellulose degradation. To examine polysaccharide-degrading potential between white- and brown-rot fungi, we performed genomewide analysis of CAZys and these oxidative enzymes in 11 Polyporales, including recently sequenced monokaryotic strains of Bjerkandera adusta, Ganoderma sp. and Phlebia brevispora. Furthermore, we conducted comparative secretome analysis of seven Polyporales grown on wood culture. As a result, it was found that genes encoding cellulases belonging to families GH6, GH7, GH9 and carbohydrate-binding module family CBM1 are lacking in genomes of brown-rot polyporales. In addition, the presence of CDH and the expansion of LPMO were observed only in white-rot genomes. Indeed, GH6, GH7, CDH and LPMO peptides were identified only in white-rot polypores. Genes encoding aldose 1-epimerase (ALE), previously detected with CDH and cellulases in the culture filtrates, also were identified in white-rot genomes, suggesting a physiological connection between ALE, CDH, cellulase and possibly LPMO. For hemicellulose degradation, genes and peptides corresponding to GH74 xyloglucanase, GH10 endo-xylanase, GH79 β-glucuronidase, CE1 acetyl xylan esterase and CE15 glucuronoyl methylesterase were significantly increased in white-rot genomes compared to brown-rot genomes. Overall, relative to brown-rot Polyporales, white-rot Polyporales maintain greater enzymatic diversity supporting lignocellulose attack.
Mycologia.Mycologia.2013 Nov-Dec;105(6):1412-27. doi: 10.3852/13-072. Epub 2013 Aug 9.
To degrade the polysaccharides, wood-decay fungi secrete a variety of glycoside hydrolases (GHs) and carbohydrate esterases (CEs) classified into various sequence-based families of carbohydrate-active enzymes (CAZys) and their appended carbohydrate-binding modules (CBM). Oxidative enzymes, such as c
PMID 23935027

Investigation of the functional role of aldose 1-epimerase in engineered cellobiose utilization.

Li S1, Ha SJ, Kim HJ, Galazka JM, Cate JH, Jin YS, Zhao H.Author information 1Energy Biosciences Institute, Institute for Genomic Biology, USA; Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Electronic address: sijinli1@illinois.edu.AbstractFunctional expression of a cellodextrin transporter and an intracellular β-glucosidase from Neurospora crassa in Saccharomyces cerevisiae enables simultaneous co-fermentation of cellobiose and non-glucose sugars such as xylose. Here we investigate the functional role of aldose 1-epimerase (AEP) in engineered cellobiose utilization. One AEP (Gal10) and two putative AEPs (Yhr210c and Ynr071c sharing 50.6% and 51.0% amino acid identity with Gal10, respectively) were selected. Deletion of GAL10 led to complete loss of both AEP activity and cell growth on cellobiose, while GAL10 complementation restored the AEP activity and cell growth. In addition, deletion of YHR210C or YNR071C resulted in improved cellobiose utilization. These results suggest that the intracellular mutarotation of β-glucose to α-glucose might be a rate controlling step and Gal10 play a crucial role in cellobiose fermentation by engineered S. cerevisiae.
Journal of biotechnology.J Biotechnol.2013 Oct 10;168(1):1-6. doi: 10.1016/j.jbiotec.2013.08.003. Epub 2013 Aug 14.
Functional expression of a cellodextrin transporter and an intracellular β-glucosidase from Neurospora crassa in Saccharomyces cerevisiae enables simultaneous co-fermentation of cellobiose and non-glucose sugars such as xylose. Here we investigate the functional role of aldose 1-epimerase (AEP) in
PMID 23954547

Japanese Journal

リン酸塩による変旋光触媒作用を利用したスクロースバイオセンサーの開発

浅見和広,稲葉英,太田口和久
化学工学論文集 36(1), 51-56, 2010-03-20
通常スクロース測定用酵素電極にはインベルターゼ（INV），ムタロターゼ（MUT），グルコースオキシダーゼ（GOD）の3種の酵素を使用するが，中でもMUTは非常に高価である．本研究では，MUTを使用しない安価な2種類の酵素から成る電極を作成し，スクロース濃度測定の可能性について検討した．グルタルアルデヒドを用いてGODを白金に架橋結合した酵素電極（GOD電極）と，GODとINVを併せて白金に架橋結合 …
NAID 10027040227

固定化酵素リアクターを用いるドリンク剤成分分析法の構築(博士論文要録)

小野真樹
分析化学 54(5), 407-408, 2005-05-05
… Enhanced interference due to excess glucose included in a pharmaceutical preparation as a sweetener was eliminated by a glucose eliminating reactor co-immobilized with three enzymes (glucose oxidase, mutarotase and catalase). …
NAID 110002910078

A Cold Acclimation Protein with Refolding Activity on Frozen Denatured Enzymes

Kawahara Hidehisa,Koda Noriko,Oshio Mika [他],OBATA Hitoshi
Bioscience, biotechnology, and biochemistry 64(12), 2668-2674, 2000-12-23
… Also, the dissociation constants, KD (M) as the binding specificity for enolase, mutarotase, isocitrate dehydrogenase, and lactate dehydrogenase were 1.82×10^<-10>, 4.35×10^<-9>, 8.98×10^<-12>, and 3.05×10^<-11>, respectively. …
NAID 110002679875

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