Thermostable Enzyme Laboratory Co.

Kōbe-shi, Japan

Thermostable Enzyme Laboratory Co.

Kōbe-shi, Japan

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Sakuraba H.,Kagawa University | Koga K.,Kagawa University | Yoneda K.,Tokai University | Kashima Y.,Thermostable Enzyme Laboratory Co. | Ohshima T.,Kyushu University
Acta Crystallographica Section F: Structural Biology and Crystallization Communications | Year: 2011

The crystal structure of an extremely thermostable multicopper oxidase (McoP) from the hyperthermophilic archaeon Pyrobaculum aerophilum was determined at a resolution of 2.0 Å. The overall fold was comprised of three cupredoxin-like domains and the main-chain coordinates of the enzyme were similar to those of multicopper oxidases from Escherichia coli (CueO) and Bacillus subtilis (CotA). However, there were clear topological differences around domain 3 between McoP and the other two enzymes: a methionine-rich helix in CueO and a protruding helix in CotA were not present in McoP. Instead, a large loop (PL-1) covered the T1 copper centre of McoP and a short -helix in domain 3 extended near the N-terminal end of PL-1. In addition, the sizes of several surface loops in McoP were markedly smaller than the corresponding loops in CueO and CotA. Structural comparison revealed that the presence of extensive hydrophobic interactions and a smaller cavity volume are likely to be the main factors contributing to the hyperthermostability of McoP. © 2011 International Union of Crystallography All rights reserved.


Patent
Thermostable Enzyme Laboratory Co. and Honda Corporation | Date: 2011-05-09

A process for producing a saccharified solution, by which xylose can be produced at a high yield from lignocellulosic biomass, is provided. The saccharification is carried out by adding a saccharifying enzyme to a pretreated material for saccharification of the lignocellulosic biomass. As the saccharifying enzyme is added a mixture of a first saccharifying enzyme originated from one or more microorganisms selected from the group consisting of fungi of the genus Acremonium, the genus Trichoderma, the genus Penicillium, the genus Aspergillus and the genus Thermoascus, and eubacteria of the genus Clostridium and the genus Bacillus, and a second saccharifying enzyme composed of -xylosidase originated from Thermotoga maritima.


Patent
Thermostable Enzyme Laboratory Co. and Totai Co. | Date: 2012-08-15

The object of the present invention is to provide a degradable composite material that contains at least one type of polysaccharide selected from chitin and chitosan, and has self-degrading capability. By supporting an enzyme capable of hydrolyzing chitin and chitosan on a molded article containing at least one type of polysaccharide selected therefrom, the polysaccharide can be slowly degraded in an environment where moisture is present such as a living body.


Nakamura T.,Japan National Institute of Advanced Industrial Science and Technology | Kashima Y.,Thermostable Enzyme Laboratory Co. | Mine S.,Japan National Institute of Advanced Industrial Science and Technology | Oku T.,Thermostable Enzyme Laboratory Co. | Uegaki K.,Japan National Institute of Advanced Industrial Science and Technology
Journal of Bioscience and Bioengineering | Year: 2012

We characterized and determined the crystal structure of a putative glucokinase/hexokinase from Thermus thermophilus that belongs to the ROK (bacterial repressors, uncharacterized open reading frames, and sugar kinases) family. The protein possessed significant enzymatic activity against glucose and mannose, with Vmax values of 260 and 68μmol·min-1·mg-1 protein, respectively. Therefore, we concluded that the enzyme is a hexokinase. However, the hexokinase showed little catalytic capacity for galactose and fructose. Circular dichroism measurements indicated that the enzyme was structurally stable at 90°C. The crystal structure of the enzyme was determined at a resolution of 2.02å, with Rcryst and Rfree values of 18.1% and 22.6%, respectively. The polypeptide structure was divided into large and small domains. The ROK consensus sequences 1 and 2 were included in the large domain. The cysteine-rich consensus sequence 2 folded into a zinc finger, and the bound zinc was confirmed by both electron density and X-ray absorption fine structure (XAFS) spectrum. The overall structure was a homotetramer that consisted of a dimer of dimers. The accessible surface area buried by the association of the dimers into the tetrameric structures was significantly higher in the T. thermophilus enzyme than in a homologous tetrameric ROK sugar kinase. © 2012 The Society for Biotechnology, Japan.


Nakamura T.,Japan National Institute of Advanced Industrial Science and Technology | Kashima Y.,Thermostable Enzyme Laboratory Co. | Mine S.,Japan National Institute of Advanced Industrial Science and Technology | Oku T.,Thermostable Enzyme Laboratory Co. | Uegaki K.,Japan National Institute of Advanced Industrial Science and Technology
Acta Crystallographica Section F: Structural Biology and Crystallization Communications | Year: 2011

Glucokinase/hexokinase catalyzes the phosphorylation of glucose to glucose 6 - phosphate, which is the first step of glycolysis. The open reading frame TTHA0299 of the extreme thermophile Thermus thermophilus encodes a putative glucokinase/hexokinase which contains the consensus sequence for proteins from the repressors, open reading frames and sugar kinases family. In this study, the glucokinase/hexokinase from T. thermophilus was purified and crystallized using polyethylene glycol 8000 as a precipitant. Diffraction data were collected and processed to 2.02 Å resolution. The crystal belonged to space group P21, with unit-cell parameters a = 70.93, b = 138.14, c = 75.16 Å, β = 95.41°. © 2011 International Union of Crystallography. All rights reserved.


Takagi K.,Konan University | Kashima Y.,Thermostable Enzyme Laboratory Co. | Fujii S.,Konan University | Koumoto K.,Konan University
Bulletin of the Chemical Society of Japan | Year: 2015

Horseradish peroxidase (HRP), a well-known oxidase, is frequently used in the diagnostic field as a labeling enzyme, where it amplifies the substrate binding signal of antibodies. Though practical for detecting moderate amounts of substrate, there is strong demand for further development of its sensitivity to detect minuscule quantities of biomarkers. Recently, we found that betaine-type cellular metabolite analogs facilitate enzymatic hydrolysis just by dissolving them into the reaction buffer. In the present study, using the analog (2-(N,N,N-tri-n-butylammonium) acetate) and various colorimetric substrates of HRP, we investigated the activation behavior of HRP. As a result, the analog structure- and concentration-dependently facilitated the various HRP-catalyzed oxidative reactions. Interestingly, the analog facilitated not only the reaction rate but also the chromogenic sensitivity. Kinetic and structural analyses revealed that the increased chromogenic sensitivity is related to the enhancement of the conformational flexibility in the substrate binding site in HRP by addition of the analog, which diminishes the binding affinity between HRP and large substrates. The finding serves to create practical applications of the analogs for increased detection sensitivity of HRP-related clinical agents. © 2015 The Chemical Society of Japan.


Patent
Thermostable Enzyme Laboratory Co. | Date: 2015-12-30

The present invention addresses the problem of providing a method for the efficient phosphorylation of glycerol. The problem is solved by reacting glycerol with either a kinase that includes the active center expressed by sequence (1) and exhibits a catalytic activity with respect to the phosphorylation of glycerol, or a kinase that includes the amino acid sequence represented by SEQ ID NO: 2 and exhibits a catalytic activity with respect to the phosphorylation of glycerol, in the presence of a phosphate group donor.


Patent
Thermostable Enzyme Laboratory Co. | Date: 2014-02-19

The present invention addresses the problem of providing a method for the efficient phosphorylation of glycerol. The problem is solved by reacting glycerol with either a kinase that includes the active center expressed by sequence (1) and exhibits a catalytic activity with respect to the phosphorylation of glycerol, or a kinase that includes the amino acid sequence represented by SEQ ID NO: 2 and exhibits a catalytic activity with respect to the phosphorylation of glycerol, in the presence of a phosphate group donor.

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