Department of Life Science, Pohang University of Science and Technology, Pohang, Republic of Korea.
Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang, Republic of Korea.
Acta Crystallogr F Struct Biol Commun. 2022 Jun 1;78(Pt 6):226-231. doi: 10.1107/S2053230X22005118. Epub 2022 May 27.
The enzymatic degradation of semi-cellulosic substrates has recently received immense attention. The enzyme endo-1,4-β-xylanase is essential for the complete digestion of complex and heterogeneous hemicellulose. Here, the purification, crystallization and preliminary X-ray free-electron laser (XFEL) diffraction analysis of endo-1,4-β-xylanase from the fungus Hypocrea virens (HviGH11) are reported. Codon-optimized HviGH11 was overexpressed in Escherichia coli and spontaneously crystallized after His-tag purification and concentration. Preliminary XFEL diffraction data were collected at the Pohang Accelerator Laboratory XFEL (PAL-XFEL). A total of 1021 images containing Bragg peaks were obtained and indexed. The HviGH11 crystals belonged to the orthorhombic space group P222, with unit-cell parameters a = 43.80, b = 51.90, c = 94.90 Å. Using 956 diffraction patterns, the phasing problem was solved and an initial model structure of HviGH11 was obtained.
最近,人们对半纤维素基质的酶促降解给予了极大的关注。内切-1,4-β-木聚糖酶对于复杂和异质的半纤维素的完全消化是必不可少的。本文报道了来自 Hypocrea virens (HviGH11) 的内切-1,4-β-木聚糖酶的纯化、结晶和初步的自由电子激光(XFEL)衍射分析。经过密码子优化的 HviGH11 在大肠杆菌中过表达,并在 His 标签纯化和浓缩后自发结晶。初步的 XFEL 衍射数据在 Pohang Accelerator Laboratory XFEL(PAL-XFEL)收集。获得并索引了总共包含布拉格峰的 1021 张图像。HviGH11 晶体属于正交晶系,空间群为 P222,晶胞参数为 a = 43.80、b = 51.90、c = 94.90 Å。使用 956 个衍射图,解决了相位问题,并获得了 HviGH11 的初始模型结构。