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643 results on '"Kuroda, Kouichi"'

12. Comparison of the mesophilic cellulosome-producing Clostridium cellulovorans genome with other cellulosome-related clostridial genomes.

Author

Tamaru, Yutaka, Miyake, Hideo, Kuroda, Kouichi, Nakanishi, Akihito, Matsushima, Chiyuki, Doi, Roy H, and Ueda, Mitsuyoshi

Subjects
Cellulosomes, Clostridium, Clostridium cellulovorans, Bacterial Proteins, Genome, Bacterial, Molecular Sequence Data, Genome Size, Genome, Bacterial, Microbiology
Abstract

Clostridium cellulovorans, an anaerobic and mesophilic bacterium, degrades native substrates in soft biomass such as corn fibre and rice straw efficiently by producing an extracellular enzyme complex called the cellulosome. Recently, we have reported the whole-genome sequence of C. cellulovorans comprising 4220 predicted genes in 5.10 Mbp [Y. Tamaru et al., (2010) J. Bacteriol., 192: 901–902]. As a result, the genome size of C. cellulovorans was about 1 Mbp larger than that of other cellulosome-producing clostridia, mesophilic C. cellulolyticum and thermophilic C. thermocellum. A total of 57 cellulosomal genes were found in the C. cellulovorans genome, and they coded for not only carbohydrate-degrading enzymes but also a lipase, peptidases and proteinase inhibitors. Interestingly, two novel genes encoding scaffolding proteins were found in the genome. According to KEGG metabolic pathways and their comparison with 11 Clostridial genomes, gene expansion in the C. cellulovorans genome indicated mainly non-cellulosomal genes encoding hemicellulases and pectin-degrading enzymes. Thus, by examining genome sequences from multiple Clostridium species, comparative genomics offers new insight into genome evolution and the way natural selection moulds functional DNA sequence evolution. Our analysis, coupled with the genome sequence data, provides a roadmap for constructing enhanced cellulosome-producing Clostridium strains for industrial applications such as biofuel production.

Published
2011

13. Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system

Author

Fukuda, Takeshi, Tsuchiyama, Kouta, Makishima, Hirokazu, Takayama, Katsumi, Mulchandani, Ashok, Kuroda, Kouichi, Ueda, Mitsuyoshi, and Suye, Shin-ichiro

Subjects
Life Sciences, Biochemistry, general, Applied Microbiology, Biotechnology, Microbiology, Cell surface engineering, Flo1p anchor system, GPI anchor system, Organophosphorus hydrolase
Abstract

Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems.

Published
2010

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