Direct visualization of Non-crystalline Cell wall Components by Ice-embedding Method.

通过冰包埋法直接观察非晶细胞壁成分。

• 批准号: 07456156
• 负责人: SUGIYAMA Junji
• 金额: $ 0.64万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07456156/
• 关键词: ice embedding; rapid-freezing; molecular chain; crystal structure; amorphous; time resoloved investigation; 経時的変化観察; 氷包埋法

It became possible to observe aqueous suspension of cellulose directly with an electron microscopy. As the preliminary results of enzymatic hydrolysis of cellulose suggest that the efficiency of degradation can be influenced by the synergism of enzymes as well as the structure of cellulose, we planned to investigate the suspension with varying periods of reaction time. However, the problems such as the gap between the cooling fin of the anticontaminator did not fit the gap of the polepiece of the existing microscope, the astigmatism induced by the power control unit prevented the high resolution observation, took us considerable time to repair the entire system. To date, most of the problems, except for the image quality, has been overcome, and we could install the best condition (40 degree tilt) for the electron diffraction analyzes of cellulose. Therefore we slightly modified our initial planning in the way to determine the molecular directionality of cellulose molecules during biosynthesis, for instance, to determine whether cellulose is synthesized from the reducing end or non-reducing end sides. So far, remarkable progress has been made on this subject, and we could show the first experimental evidences that the reducing-end of cellulose molecule points outward the organism, in another word, the addition of monomer occurs at the non-reducing end of the preexisting cellulose molecule.

用电子显微镜直接观察纤维素的水悬浮液成为可能。由于纤维素酶水解的初步结果表明，降解效率可以受到酶的协同作用以及纤维素的结构的影响，我们计划研究不同反应时间的悬浮液。然而，由于防污染器散热片之间的差距与现有显微镜极镜的差距不匹配，功率控制单元引起的像散妨碍了高分辨率观察，我们花费了相当长的时间来修复整个系统。到目前为止，除了图像质量之外，大多数问题都已得到解决，我们可以为纤维素的电子衍射分析设置最佳条件（40度倾斜）。因此，我们稍微修改了我们最初的计划，以确定纤维素分子在生物合成过程中的分子方向性，例如，确定纤维素是从还原端还是非还原端合成的。迄今为止，这方面的研究已取得了显著进展，我们首次从实验上证明了纤维素分子的还原端指向生物体外，即单体的加成发生在纤维素分子的非还原端。

项目成果

Okamoto, K.: "Structural Diversity of Ascidian Crystalline Cellulose" Wood Research. 83. 27-29 (1996)

Okamoto, K.：“海鞘结晶纤维素的结构多样性”木材研究。

Okamoto, K.et al.: "Structural diversity of Ascidian crystalline cellulose." Wood Research. 83. 27-29 (1996)

Okamoto, K.et al.：“海鞘结晶纤维素的结构多样性。”

Samejima, M.et al.: "The behavior of Phanerochaete chrysosporium cellobiose dehydrogenaze on adsorption to crystalline and amorphous cellulose." Biotechnol.Appl.Biochem.25, (in press). (1997)

Samejima, M.et al.：“Phanerochaete chrysosporium 纤维二糖脱氢酶对结晶和无定形纤维素的吸附行为。”

Ishikawa, A.et al.: "Fine structure and tensile properties of ramie fibers in the crystalline form of cellulose I,II,III_I and IV_I." Polymer. 38-2. 463-468 (1997)

Ishikawa, A. 等人：“纤维素 I、II、III_I 和 IV_I 结晶形式的苎麻纤维的精细结构和拉伸性能。”

Wada, M.ら: "Synchrotron-radiated X-ray and neutron diffraction study of native cellulose" Cellulose. 4. (1997)

Wada, M. 等人：“天然纤维素的同步辐射 X 射线和中子衍射研究”纤维素 4。（1997 年）