Novel technique for monitoring enzymatic hydrolysis of cellulose through high throughput DP analysis

• Project/Area Number: 25660138
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Wood science
• Research Institution: Kyoto University
• Principal Investigator: SUGIYAMA Junji 京都大学, 生存圏研究所, 教授 (40183842)
• Co-Investigator(Kenkyū-buntansha): HORIKAWA Yoshiki 京都大学, 生存圏研究所, 特定研究員 (90637711)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ケモメトリクス / 木質バイオマス / 分子量 / ハイスループット / セルロース / セルラーゼ / 近赤外分光分析 / 近赤外スペクトロスコピー / 糖化 / エンドグルカナーゼ活性 / 重合度 / 多変量解析

Outline of Final Research Achievements

In order to improve saccharification performance, it is important to obtain precise information about functions and activities of individual enzymes in a cocktail of the cellulase. Particularly, endo-activity conventionally estimated by digestion of CMC, a model amorphous compound, did not often conform to the degradation of pretreated biomass. Since the accurate characterization of cellulose chain length seemed critical to the measure of endo-performance, highly dispersed microfibrillated cellulose samples were prepared as a standard substrate and the degree of polymerization (DP) was precisely monitored by GPC after tricarbanilation of cellulose. Surprisingly, for the first time, endo-activity of EG II was found to possess higher than that of EG I, while no considerable differences were found in those between CBH II and CBH I. Furthermore, facile and quick chemometrical monitoring system of DP was successfully developed using Near Infrared (NIR) spectra from the hydrolysates

Research Products

• [Journal Article] Line monitoring by near-infrared chemometric technique for potential ethanol production from hydrothermally treated Eucalyptus globulus. (2015)
  • Author(s): Horikawa, Y., Imai, M., Kanai, K., Imai, T., Watanabe, T., Takabe, K., Kobayashi, Y., Sugiyama, J.
  • Journal Title: Biochemical Engineering Journal Volume: 97 Pages: 65-72
  • DOI: 10.1016/j.bej.2015.01.003
  • NAID: 120005555522
  • Peer Reviewed
• [Journal Article] Near-infrared spectroscopy as a potential method for identification of anatomically similar Japanese diploxylons. (2015)
  • Author(s): Horikawa, Y., Mizuno-Tazuru, S., Sugiyama, J.
  • Journal Title: Journal of Wood Science Volume: 61 Issue: 3 Pages: 251-261
  • DOI: 10.1007/s10086-015-1462-2
  • NAID: 120005669339
  • Peer Reviewed / Open Access
• [Presentation] 透過型電子顕微鏡によるセルラーゼ-セルロース相互作用の可視化 (2015)
  • Author(s): 堀川祥生、今井友也、杉山淳司
  • Organizer: 第65回日本木材学会大会
  • Place of Presentation: タワーホール船堀(東京都江戸川区）
  • Year and Date: 2015-03-16 – 2015-03-18
• [Presentation] Near-infrared spectroscopy as a potential method for identification of anatomically similar Japanese diploxylons (2014)
  • Author(s): Horikawa Y., Mizuno-Tazuru S., Sugiyama J
  • Organizer: 9th Joint Seminar of China-Korea-Japan on Wood Quality and Utilization of Domestic Species
  • Place of Presentation: 秋田ビューホテル（秋田県秋田市）
  • Year and Date: 2014-10-29 – 2014-10-30
• [Presentation] TEMによる酵素-基質相互作用の可視化 (2014)
  • Author(s): 堀川祥生、今井友也、杉山淳司
  • Organizer: セルロース学会第21回年次大会
  • Place of Presentation: 鹿児島大学稲盛会館(鹿児島県鹿児島市）
  • Year and Date: 2014-07-17 – 2014-07-18
• [Presentation] 透過型電子顕微鏡を用いた酵素-基質相互作用の可視化 (2014)
  • Author(s): 堀川祥生、今井友也、杉山淳司
  • Organizer: セルラーゼ研究会第28回大会
  • Place of Presentation: 幕張セミナーハウス(千葉県習志野市)
  • Year and Date: 2014-07-11 – 2014-07-12
• [Presentation] セルロースの重合度測定によるエンドグルカナーゼ活性評価法の開発とそのハイスループット解析 (2014)
  • Author(s): 堀川祥生, 阿部賢太郎, 今井友也, 榊原圭太, 辻井敬亘, 小林良則, 杉山淳司
  • Organizer: 第64回日本木材学会大会
  • Place of Presentation: 松山、愛媛
• [Presentation] 重合度測定によるセルラーゼ活性評価法の開発とそのハイスループット化 (2013)
  • Author(s): 堀川祥生, 阿部賢太郎, 今井友也, 榊原圭太, 辻井敬亘, 小林良則, 杉山淳司
  • Organizer: 第20回日本セルロース学会年次大会
  • Place of Presentation: 宇治、京都
• [Presentation] Practical evaluation for cellulase activity by estimating degree of polymerization (DP) of cellulose and its high-throughput analysis. (2013)
  • Author(s): Yoshiki Horikawa, Tomoya Imai, Junji Sugiyama, Keita Sakakibara, Yoshinobu Tsujii, Yoshinori Kobayashi
  • Organizer: th Pacific Regional Wood Anatomy Conference and Annual Meeting of International Academy of Wood Science
  • Place of Presentation: Nanjing, China
• [Presentation] Assessment of endoglucanase activity by analyzing degree of polymerization (DP) of cellulose and its high-throughput analysis. (2013)
  • Author(s): Yoshiki Horikawa, Tomoya Imai, Junji Sugiyama, Keita Sakakibara, Yoshinobu Tsujii, Yoshinori Kobayashi
  • Organizer: International Symposium on Wood Quality and Utilization
  • Place of Presentation: Jeju, Korea