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Kamino K.,Marine Biotechnology Institute | Kamino K.,National Institute of Technology and Evaluation | Nakano M.,Marine Biotechnology Institute | Kanai S.,Pharma Design Inc.
FEBS Journal | Year: 2012

Barnacles are a unique sessile crustacean that attach irreversibly and firmly to foreign underwater surfaces. Its biological underwater adhesive is a peculiar extracellular multi-protein complex. Here we characterize one of the two major proteins, a 52 kDa protein found in the barnacle cement complex. Cloning of the cDNA revealed that the protein has no homolog in the nonredundant database. The primary structure consists of four long sequence repeats. The process of dissolving the protein at the adhesive joint of the animal by various treatments was monitored in order to obtain insight into the molecular mechanism involved in curing of the adhesive bulk. Treatments with protein denaturant, reducing agents and/or chemical-specific proteolysis in combination with 2D diagonal PAGE indicated no involvement of the protein in intermolecular cross-linkage/polymerization, including formation of intermolecular disulfide bonds. As solubilization of the proteins required high concentrations of denaturing agents, it appears that both the conformation of the protein as building blocks and non-covalent molecular interactions between the building blocks, possibly hydrophobic interactions and hydrogen bonds, are crucial for curing of the cement. It was also suggested that the protein contributes to surface coupling by an anchoring effect to micro- to nanoscopic roughness of surfaces. Database -Sequence of Megabalanus rosa cp52k mRNA for 52 kDa cement protein has been submitted to the DNA Data Bank of Japan under accession number. Barnacle attaches irreversibly and firmly to foreign underwater surfaces. Characterization of the major bulk protein at the adhesive joint of the animal indicated no involvement of the protein in the intermolecular cross-linkage/polymerization. It appeared that both conformation of the protein and non-covalent molecular interactions, possibly hydrophobic interactions and hydrogen bonds, are crucial for curing of the adhesive. © 2012 The Authors Journal compilation © 2012 FEBS.


Kamino K.,National Institute of Technology and Evaluation
Biofouling | Year: 2013

Barnacles are intriguing, not only with respect to their importance as fouling organisms, but also in terms of the mechanism of underwater adhesion, which provides a platform for biomimetic and bioinspired research. These aspects have prompted questions regarding how adult barnacles attach to surfaces under water. The multidisciplinary and interdisciplinary nature of the studies makes an overview covering all aspects challenging. This mini-review, therefore, attempts to bring together aspects of the adhesion of adult barnacles by looking at the achievements of research focused on both fouling and adhesion. Biological and biochemical studies, which have been motivated mainly by understanding the nature of the adhesion, indicate that the molecular characteristics of barnacle adhesive are unique. However, it is apparent from recent advances in molecular techniques that much remains undiscovered regarding the complex event of underwater attachment. Barnacles attached to silicone-based elastomeric coatings have been studied widely, particularly with respect to fouling-release technology. The fact that barnacles fail to attach tenaciously to silicone coatings, combined with the fact that the mode of attachment to these substrata is different to that for most other materials, indicates that knowledge about the natural mechanism of barnacle attachment is still incomplete. Further research on barnacles will enable a more comprehensive understanding of both the process of attachment and the adhesives used. Results from such studies will have a strong impact on technology aimed at fouling prevention as well as adhesion science and engineering. © 2013 Copyright Taylor and Francis Group, LLC.


Tanasupawat S.,Chulalongkorn University | Kommanee J.,Chulalongkorn University | Yukphan P.,National Science and Technology Development Agency | Nakagawa Y.,National Institute of Technology and Evaluation | And 3 more authors.
Journal of the Science of Food and Agriculture | Year: 2011

Background: Fermented rice flour (khao-khab, a non-glutinous rice) and related products are Thai traditional products. The types of acetic acid bacteria (AAB) microflora in khao-khab have not been reported. In this study, Acetobacter strains were isolated and identified based on the phenotypic and chemotaxonomic characteristics and molecular aspects. Results: Twenty-five acetic acid bacteria isolated from fermented rice products and a starter for sweetened rice in Thailand by an enrichment culture approach, were assigned to the genus Acetobacter by phenotypic and chemotaxonomic characterisations. On the basis of the 16S rRNA gene sequence and 16S-23S rRNA gene ITS restriction analyses, 25 isolates were divided into six groups and identified at the specific level: (1) Group 1 included five isolates, which were identified as A. indonesiensis; (2) Group 2 included two isolates, which were identified as A. lovaniensis; (3) Group 3 included one isolate, which was identified as A. orientalis; (4) Group 4 included eleven isolates, which were identified as A. pasteurianus; (5) Group 5 included three isolates, which were identified as A. syzygii and (6) Group 6 included three isolates, which were unidentified and considered to constitute a new species. Conclusion: Results revealed that various Acetobacter species were distributed in Thai fermented rice flour and related products. A novel Acetobacter species was isolated from the product. © 2011 Society of Chemical Industry.


Sukkhum S.,Kasetsart University | Sukkhum S.,Srinakharinwirot University | Tokuyama S.,University of Shizuoka | Kongsaeree P.,Kasetsart University | And 3 more authors.
African Journal of Microbiology Research | Year: 2011

An actinomycete strain T16-1 demonstrated the maximum poly (L-Lactide)-degrading activity when cultured in basal liquid medium at 50°C. According to 16S rDNA sequence analysis, chemotaxonomic and DNA-DNA hybridization revealed that strain T16-1 belong to the family Thermomonosporaceae, genus Actinomadura. On the basis of phenotypic and phylogenic data, strain T16-1 which is a novel PLA-degrading thermophilic actinomycete was identified as Actinomadura keratinilytica, but the color of its colony on ISP plates, NaCl tolerant and utilization of mannitol, raffinose and arabinose were different. Partial sequence of poly (L-lactide) depolymerase gene from strain T16-1 was demonstrated. The gene consisting of 222 amino acids was related to serine protease from Streptomyces sp. with 43 to 46% identity. © 2011 Academic Journals.


Yamamura H.,Yamanashi University | Yamamura H.,National Institute of Technology and Evaluation | Lisdiyanti P.,Indonesian Institute of Sciences | Ridwan R.,Indonesian Institute of Sciences | And 7 more authors.
International Journal of Systematic and Evolutionary Microbiology | Year: 2010

An actinomycete strain, ID05-A0528T, was isolated using the SDS-yeast extract pre-treatment method from soil under mahogany (Swietenia mahogani) trees in West Timor, Indonesia, and was examined by using a polyphasic taxonomic approach. Chemotaxonomic and phylogenetic characterizations demonstrated that the novel strain belongs to the genus Dietzia. 16S rRNA gene sequencing studies showed that the strain was related to Dietzia cinnamea (97.2 %). Results of phenotypic and phylogenetic analyses determined that strain ID05-A0528T is different from the known species of the genus Dietzia. It is proposed that the isolate should be classified as a representative of a novel species of the genus Dietzia, with the name Dietzia timorensis sp. nov. The type strain is ID05-A0528T (=BTCC B-560T =NBRC 104184T). © 2010 IUMS.

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