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36 results on '"Kusaoke, Hideo"'

2. Chromosomal structure of the halophilic archaebacterium Halobacterium salinarium

Author

Takayanagi, Shinnosuke, Morimura, Satoshi, Kusaoke, Hideo, Yokoyama, Yoshiko, Kano, Kazutaka, and Shioda, Masaki

Subjects
Bacteria -- Genetic aspects, Bacterial genetics -- Research, Biological sciences
Abstract

The structure of the chromosome of the halophilic archaebacterium Halobacterium salinarium was examined. Sucrose gradient centrifugation and electron microscopic observations revealed that the bacterial chromosome consists of regions of both protein-free DNA and protein-associated DNA. The latter region was composed of nucleosome-like structures that form a high-order fibrous structure. Moreover, these regions are changeable depending on the growth phase.

Published
1992

7. An Improved Method for DNA Transformation of Intact Cells of Bacillus subtilis by Electroporation

Author

KUSAOKE, Hideo and KADOWAKI, Yasuyuki

Abstract

An electroporation procedure for the plasmid-mediated genetic transformation of intact cells of Bacillus subtilis vas performed. The method was optimized for B. subtilis ISWl214,and the transformation of ISW1214 , with plasmid pUB110,using the mixture of 0.3 M sucrose I mM CaCl_2 and 1 DM sodium citrate as a buffer for pulsation, results in a maximum efficiency of 2.8 x 104 transfornants per μg DNA, with a single pulse duration of 500 μsec with an initial electric field strength of 7 kV/cm. Transformation of ISW1214 occurred at comparable frequencies with pUB110(4.5 kbp), pC194(2.9 kbp), and pE194(3.7 kbp) plasmids. Plasmid DNA isolated from transformants , , as indistinguishable from authentic preparation of plasmid DNA on gel electrophoretic analysis. The electroporation was simple, rapid, and sampled at 10 μ1-small volume of cell suspension(～IxlO^ cells/ml, 4 pg DNA/μl).

Published
1992

10. Special feature

Author

Kimoto, Hisashi, primary, Itoh, Takafumi, additional, Hibi, Takao, additional, Fujii, Yutaka, additional, and Kusaoke, Hideo, additional

Published
2014
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21. Discoidin Domain of Chitosanase Is Required for Binding to the Fungal Cell Wall.

Author

Kimoto, Hisashi, Akamatsu, Miho, Fujii, Yutaka, Tatsumi, Hirosuke, Kusaoke, Hideo, and Taketo, Akira

Subjects
HYDROLASES, ENZYMES, ZYGOMYCETES, CHITIN, CHITOSAN, GLUCANS
Abstract

Previously, we reported properties of a glycosylase belonging to GH-8 glycosyl hydrolase (GH) and having both chitosanase and glucanase activities. This enzyme (D2), whose molecular mass (86 kDa) was the largest among the GH-8 group, has its catalytic domain at the N-terminal region, and discoidin domain (DD) at the C-terminal region. Although various chitosanases, chitinases and glucanases have been known, DD is unique to the D2 enzyme. Glucanase and chitinase, but not chitosanase, are known to have functional domain such as carbohydrate-binding module, besides catalytic domain. Accordingly, function of the DD of D2 chitosanase was analyzed, using zygomycete cell wall containing chitosan, glucan and chitin as the basic constituents. The DD specifically and tightly bound to chitosan, but did not participate in affinity for glucan and chitin. Deletion of the DD caused marked reduction in absorbability to cell wall and in hydrolytic activity toward chitosan and glucan. These results suggest that the DD is concerned in binding of the enzyme to cell wall and in effective digestion of the insoluble substrate, through hydrolysis of not only chitosan but also coexisting glucan. Thus, this is the first example of chitosan-binding domain among various carbohydrate-binding modules reported thus far. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Published
2010
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25. Immobilization of Invertase on Carboxymethylchitosan and Chitosan

Author

KUSAOKE, Hideo

Abstract

An immobilization method using carboxymethylchitosan prepared from chitosan as an insoluble carrier was investigated. Invertase was immobilized on carboxymethylchitosan using the bifunctional agent, glutaraldehyde. The most active preparation was based on the enzyme bound carboxymethylchitosan with low degree of substitution (d.s.) such as 0.41. Its activity yield was 24.1%. The activity of the immobilized enzyme decreased as the d.s. of carboxymethyl groups increased. Compared with free invertase, immobilized enzyme exhibited similar pH optimum but more activity at the range of pH 5-7. Immobilized enzyme improved thermal stability. The immobilized enzyme retained ca. 70% of the initial activity even after the repeated use of 10 times.

Published
1988

26. Immobilization of α-Chymotrypsin and Other Enzymes on Acylchitosan

Author

KUSAOKE, Hideo

Abstract

α-Chymotrypsin has been immobilized on acylchitosan, a chitosan derivative, using glutaraldehyde as an intermediate reagent ; the activity of the immobilized enzyme on acylchitosan was much higher than that on chitosan. The increase of the degree of substitution of acyl groups in acylchitosan as support was effective for the enzyme activity. The retained maximum activity of immobilized α-chymotrypsin was approximately 60% that of the native enzyme, obtained by the use of the butyrylchitosan with the degree of substitution of butyryl groups of 0.87. Immobilized α-chymotrypsin showed a significant changes in optimal temperature and heat stability. The pH optimum of the fixed α-chymotrypsin was simillar to that of native enzyme. Immobilized α-amylase and invertase on acylchitosan also showed a slight higher enzyme activity than that on chitosan.

Published
1986

27. Yeast Cell-Surface Expression of Chitosanase from Paenibacillus fukuinensis.

Author

Fukuda, Takeshi, Isogawa, Danya, Takagi, Madoka, Kato-Murai, Michiko, Kimoto, Hisashi, Kusaoke, Hideo, Ueda, Mitsuyoshi, and Suye, Shin-ichiro

Subjects
ENZYMES, YEAST, SACCHARIDES, CELLS, IMMUNOFLUORESCENCE
Abstract

The article discusses a study which examines yeast cell-surface expression of chitosanase from Paenibacillus fukuinensis. To produce chitoorigosaccharides using chitosan, Paenibacillus fukuinensis chitosanase-displaying yeast cells were constructed as a whole-cell biocatalyst through yeast cell-surface engineering. Immunofluorescence labeling of cells confirmed the localization of the chitosanase on the yeast cell surface.

Published
2007
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35. Effects of DNA Topology on Transformation Efficiency of Bacillus subtilisISW1214 by Electroporation

Author

Ohse, Morimasa, Kawade, Koji, and Kusaoke, Hideo

Abstract

We report an investigation of electrotransformation by three different topological isomers, circular supercoiled (sc DNA), circular relaxed (cr DNA), and linearized (In DNA) forms of the plasmids pUB110 (4.5 kbp) and pBDR331T (12.6 kbp), of a Gram-positive bacterium, Bacillus subtilisISW1214. Treatment of the sc DNA with calf thymus topoisomerase I removed the superhelicity and the DNA assumed the relaxed circular form. Treatment of sc DNA with restriction endonuclease linearized the DNA. The transformation with the sc DNA of pUB110 resulted in the maximum efficiency of (2.6±0.6) × 105transformants per μg DNA higher than that ((2.0±0.3) × 104transformants per μg DNA) for the cr DNA, using the DNA concentration of 20 μg/ml at an electric field strength of 7kV/cm and a capacitance of 10 μF with a single decayed pulse. The transformation efficiency (TE) for the In DNA was zero. The variations of TE for different topological forms of DNA reflected their relative stability in the host cells. The molecular efficiency (ME, transformants per molecule) for sc DNA was nearly one order of magnitude greater for the lower molecular size of pUB110 DNA than that for the higher molecular size of pBDR331T DNA.

Published
1997
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36. Biochemical and genetic properties of Paenibacillus glycosyl hydrolase having chitosanase activity and discoidin domain.

Author

Kimoto H, Kusaoke H, Yamamoto I, Fujii Y, Onodera T, and Taketo A

Subjects
Amino Acid Sequence, Base Sequence, Blotting, Northern, Blotting, Western, Catalytic Domain, Cloning, Molecular, DNA, Bacterial, Discoidins, Glycoside Hydrolases chemistry, Glycoside Hydrolases genetics, Molecular Sequence Data, Mutagenesis, Site-Directed, Protozoan Proteins chemistry, Protozoan Proteins genetics, Sequence Homology, Amino Acid, Bacillus enzymology, Glycoside Hydrolases metabolism, Lectins, Protozoan Proteins metabolism
Abstract

Cells of "Paenibacillus fukuinensis" D2 produced chitosanase into surrounding medium, in the presence of colloidal chitosan or glucosamine. The gene of this enzyme was cloned, sequenced, and subjected to site-directed mutation and deletion analyses. The nucleotide sequence indicated that the chitosanase was composed of 797 amino acids and its molecular weight was 85,610. Unlike conventional family 46 chitosanases, the enzyme has family 8 glycosyl hydrolase catalytic domain, at the amino-terminal side, and discoidin domain at the carboxyl-terminal region. Expression of the cloned gene in Escherichia coli revealed beta-1,4-glucanase function, besides chitosanase activity. Analyses by zymography and immunoblotting suggested that the active enzyme was, after removal of signal peptide, produced from inactive 81-kDa form by proteolysis at the carboxyl-terminal region. Replacements of Glu(115) and Asp(176), highly conserved residues in the family 8 glycosylase region, with Gln and Asn caused simultaneous loss of chitosanase and glucanase activities, suggesting that these residues formed part of the catalytic site. Truncation experiments demonstrated indispensability of an amino-terminal region spanning 425 residues adjacent to the signal peptide.

Published
2002
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