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Comparative Biochemical Analysis of Cellulosomes Isolated from Clostridium clariflavum DSM 19732 and Clostridium thermocellum ATCC 27405 Grown on Plant Biomass

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Abstract

The cellulosome is a supramolecular multienzyme complex formed via species-specific interactions between the cohesin modules of scaffoldin proteins and the dockerin modules of a wide variety of polysaccharide-degrading enzymes. Here, we report a comparative analysis of cellulosomes prepared from the thermophilic anaerobic bacteria Clostridium (Ruminiclostridium) clariflavum DSM 19732 and Clostridium (Ruminiclostridium) thermocellum ATCC 27405 grown on delignified rice straw. The results indicate that the isolated C. clariflavum cellulosome exhibits lower activity for insoluble cellulosic substrates and higher activity for hemicellulosic substrates, especially for xylan, compared to the isolated C. thermocellum cellulosome. The C. clariflavum cellulosome was separated into large and small complexes by size exclusion chromatography, and the high xylanase activity of the intact complex is mainly attributed to the small complex. Furthermore, both C. clariflavum and C. thermocellum cellulosomes efficiently converted delignified rice straw into soluble sugars with different compositions, whereas a mixture of these cellulosomes exhibited essentially no synergy for the saccharification of delignified rice straw. This is the first study to report that isolated C. clariflavum cellulosomes exhibit greater xylanase activity than isolated C. thermocellum cellulosomes. We also report the effect of a combination of intact cellulosome complexes isolated from different species on the saccharification of plant biomass.

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Funding

This work was funded by PRESTO grant JPMJPR12B7 from the Japan Science and Technology Agency and a grant (26550103) for Challenging Exploratory Research and a Grant-in-Aid for Scientific Research (B) (16H04732) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. This research was also funded by grants for Strategic Research Foundations at Private Universities (2013–2017 and 2014–2018) from MEXT, Japan.

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Authors and Affiliations

  1. Department of Chemical Biology & Applied Chemistry, College of Engineering, Nihon University, 1 Nakagawara, Tokusada, Tamura-machi, Koriyama, Fukushima, 963-8642, Japan

    Suguru Shinoda, Masahiro Kurosaki, Takaaki Kokuzawa, Katsuaki Hirano, Mitsuru Haruki & Nobutaka Hirano

  2. Department of Applied Biological Science, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa, 252-0880, Japan

    Hatsumi Takano & Kenji Ueda

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  1. Suguru Shinoda
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  2. Masahiro Kurosaki
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Correspondence to Nobutaka Hirano.

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Shinoda, S., Kurosaki, M., Kokuzawa, T. et al. Comparative Biochemical Analysis of Cellulosomes Isolated from Clostridium clariflavum DSM 19732 and Clostridium thermocellum ATCC 27405 Grown on Plant Biomass. Appl Biochem Biotechnol 187, 994–1010 (2019). https://doi.org/10.1007/s12010-018-2864-6

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