To Investigate the Assembly Mechanism of Recombinant Spider Dragline Silk Proteins through In-situ Crystallization in a Biomimetic Microfluidic Chip

通过仿生微流控芯片中的原位结晶研究重组蜘蛛拖丝蛋白的组装机制

• 批准号: 21K15063
• 负责人: Chen Jianming
• 金额: $ 3万
• 依托单位: Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K15063/
• 关键词: Spider dragline silk; Assembly mechanism; Recombinant technology; Crystallization; Microfluidic chip

In the research plan, the first step is to prepare the genetically engineered spider silk proteins. Currently, we have designed six mutants and successfully purified these proteins. A biomimetic microfluidic chip was made to mimic the natural spinning process, allowing the introduction of all triggers to the channel in order. It is flexible to change the conditions (such as concentration, pH and negative pressure) and totally different silk morphologies can be found along the channel. Various microscopes were used to observe the surface structure of silk morphologies along the channel. Interestingly, the beta-sheet formation was well characterized by Raman and synchrotron radiated WAXD. Now we are summarizing the data and preparing the manuscript to publish this result soon.

在研究计划中，第一步是制备基因工程蜘蛛丝蛋白。目前，我们已经设计了六个突变体，并成功地纯化了这些蛋白。一个仿生微流控芯片被用来模仿自然的旋转过程，允许将所有的触发器按顺序引入通道。该方法可以灵活地改变条件（如浓度、pH值和负压），沿着通道可以发现完全不同的丝形态。使用各种显微镜观察丝形态沿着通道的表面结构。有趣的是，β-片层的形成，以及其特征在于拉曼和同步辐射WAXD。现在我们正在汇总数据，准备手稿，以便尽快发表这一结果。

项目成果

Preparation of Extensible Silk Fibers in an Aqueous Citrate Buffer System

柠檬酸盐缓冲液体系中可拉伸丝纤维的制备

• 发表时间: 2021
• 作者: Jianming Chen;Yoshinori Ohta;Hiroyuki;Nakamura;Hiroyasu Masunaga;Keiji Numata
• 通讯作者: Keiji Numata

Silk Composite Fibers Reinforced by the Telechelic Polyalanine

遥爪聚丙氨酸增强丝复合纤维

• 发表时间: 2021
• 作者: Jianming Chen;Kousuke Tsuchiya;Hiroyasu Masunaga;Ali D. Malay;Keiji Numata
• 通讯作者: Keiji Numata