Analysis of microbial machinery for excretion by using patch-clamp method and creation of novel excretion system

• Project/Area Number: 24580124
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Tokyo Denki University
• Principal Investigator: KAWASAKI Hisashi 東京電機大学, 工学部, 教授 (90349788)
• Co-Investigator(Renkei-kenkyūsha): HASHIMOTO Ken-ichi 東京電機大学, 工学部, 助教 (70508382)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 排出 / 代謝工学 / 合成生物学 / 発酵 / Corynebacterium / チャネル / パッチクランプ法 / 応用微生物 / 電気生理

Outline of Final Research Achievements

While it is hoped that microbial cell factories can be created to produce various useful compounds, it is thought unlikely that a pre-existing natural exporter exists that can selectively export all of these useful compounds out of the cell. In addition, rational design of such exporters is, at present, extremely difficult. This is thought to be one cause of the inability of designed microbial cell factories to perform as expected, and there is demand for a breakthrough for this problem.
In this study, having confirmed the export activity of actual gene product using our unique path-clamp system, we introduced exporter genes into microbial cell factories, and showed that exporter engineering is highly important in the creation of microbial cell factories by systems biology or synthetic biology.

Research Products

• [Journal Article] Glutamate Is Excreted Across the Cytoplasmic Membrane through the NCgl1221 Channel of <i>Corynebacterium glutamicum</i> by Passive Diffusion (2012)
  • Author(s): Ken-ichi Hashimoto, Jun Murata, Tomoyuki Konishi, Isamu Yabe, Tsuyoshi Nakamatsu, and Hisashi Kawasaki
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 76 Issue: 7 Pages: 1422-1424
  • DOI: 10.1271/bbb.120366
  • NAID: 10030982446
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Presentation] メカノセンシティブチャネル遺伝子のリジン生産菌への導入効果 (2015)
  • Author(s): 三井 岳志，小西 智之，橋本 賢一，矢部 勇，川崎 寿
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学（岡山県岡山市）
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] 分子動力学シミュレーションに支援されたTween 40によるCorynebacterium glutamicumのグルタミン酸過剰生成誘導機構の解析 (2015)
  • Author(s): 澤田 康之，橋本 賢一，中松 亘，曽我部 正博，川崎 寿
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学（岡山県岡山市）
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] Effects of introducing a mechanosensitive channel gene from Corynebacterium glutamicum into Escherichia coli (2014)
  • Author(s): Tomoyuki Konishi, Ken-ichi Hashimoto, Isamu Yabe, Hisashi Kawasaki
  • Organizer: Mechanosensory Transduction Satellite Symposium in conjunction with IUPAB2014
  • Place of Presentation: Gold Coast, Australia
  • Year and Date: 2014-07-30 – 2014-08-02
• [Presentation] Applicability of mechanosensitive channel in Corynebacterium glutamicum as a versatile exporter (2014)
  • Author(s): Ken-ichi Hashimoto, Tomoyuki Konishi, Isamu Yabe, Hisashi Kawasaki
  • Organizer: 16th European Congress on Biotechnology
  • Place of Presentation: Edinburgh, Scotland
  • Year and Date: 2014-07-13 – 2014-07-16
• [Presentation] Mechanosensitive channels as potentiallly versatile exporters
  • Author(s): Ken-ichi Hashimoto, Tomoyuki Konishi, Isamu Yabe, Tsuyoshi Nakamatsu, Hisashi Kawasaki
  • Organizer: Society for industrial microbiology and biotechnology annual meeting
  • Place of Presentation: Sheraton San Diego （Dan Diego, CA)
• [Presentation] L-Glutamate excretion through the NCgl1221 channel and its potential as a tool for exporter engineering
  • Author(s): Hisashi Kaswasaki
  • Organizer: Methods for the investigation of ion transport machinery in biological membranes
  • Place of Presentation: Italian Institute of Culture (Chiyoda-ku, Tokyo)
  • Invited
• [Presentation] 汎用型物質排出担体としてのCorynebacterium glutamicum由来圧力依存性チャネルの有用性
  • Author(s): 橋本 賢一、小西 智之、李 双春、三井 岳志、中松 亘、川崎 寿
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 明治大学生田キャンパス（神奈川県）
• [Presentation] Corynebacterium glutamicum由来のグルタミン酸排出チャネルであるNCgl1221遺伝子産物のＣ末端ドメインの電気生理学的手法を用いた機能解析
  • Author(s): 橋本 賢一，山下 周子，北嶋 俊一，矢部 勇，夏目 亮，中松 亘，川崎 寿，和地 正明
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学
• [Book] 化学と生物 (2012)
  • Author(s): 川崎 寿，和地 正明
  • Total Pages: 9
  • Publisher: 学会出版センター
• [Patent(Industrial Property Rights)] プリン系物質の製造法 (2013)
  • Inventor(s): 川崎 寿、橋本 賢一、中松 亘、浅原 貴之
  • Industrial Property Rights Holder: 味の素株式会社
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-161662
  • Filing Date: 2013-08-02