Preparation of Large-Volume Crystal of Cellulase Under Microgravity to Investigate the Mechanism of Thermal Stabilization
Preparation of Large-Volume Crystal of Cellulase Under Microgravity to Investigate the Mechanism of Thermal Stabilization
复制标题
微重力下制备大体积纤维素酶晶体以研究热稳定机制
DOI：
10.15011/jasma.38.1.380103
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
K. Igarashi
中科院分区：
文献类型：
--
作者：
Sora Yamaguchi;Naoki Sunagawa;K. Matsuyama;M. Tachioka;E. Hirota;S. Takahashi;K. Igarashi
纤维素酶对纤维素的酶催化糖化有望使生物质中可溶糖的高效生产成为可能，因此纤维素酶的催化机制已经得到了深入的研究。我们已经证明，从担子菌Phanerochaete chrassporium中获得的纤维生物水解酶Cel6A的非催化半胱氨酸残基突变改善了酶的热稳定性。为了理解其中的原因，我们考虑用中子衍射分析来可视化突变酶内部的氢键网络。在本研究中，我们首先考察了制备大体积晶体所需的NaCl和聚乙二醇的最佳浓度范围，并在此基础上选择了国际空间站微重力条件下的9组结晶条件。成功地获得了大体积的突变酶晶体。
Enzymatic saccharification of cellulose by cellulases is expected to enable energy-efficient production of soluble sugars from biomass, and thus the catalytic mechanisms of cellulases have been intensively studied. We have shown that mutation of non-catalytic cysteine residues of cellobiohydrolase Cel6A from the basidiomycete Phanerochaete chrysosporium improves the thermal stability of the enzyme. To understand why, we considered visualizing the hydrogen bond network within the mutant enzyme by neutron diffraction analysis. In this study, we first examined the optimum concentration ranges of NaCl and polyethylene glycol to produce the required large-volume crystal, and based on the results, we chose nine sets of conditions for crystallization under microgravity at the International Space Station. A large-volume crystal of the mutant enzyme was obtained successfully.
DOI：
10.1107/s0108767307043498
发表时间：
2008
期刊：
Acta crystallographica. Section A, Foundations of crystallography
影响因子：
--
作者：
N. Niimura;R. Bau
通讯作者：
N. Niimura;R. Bau
DOI：
10.1107/s2059798319011471
发表时间：
2019-10-01
影响因子：
2.2
作者：
Liebschner, Dorothee;Afonine, Pavel V.;Adams, Paul D.
通讯作者：
Adams, Paul D.
影响因子：
3.8
作者：
Inaka, Koji;Takahashi, Sachiko;Aritake, Kosuke;Tsurumura, Toshiharu;Furubayashi, Naoki;Yan, Bin;Hirota, Erika;Sano, Satoshi;Sato, Masaru;Kobayashi, Tomoyuki;Yoshimura, Yoshinori;Tanaka, Hiroaki;Urade, Yoshihiro
通讯作者：
Urade, Yoshihiro