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来自绿色木霉内生 1,4-β-木聚糖酶晶体的初步 XFEL 数据。

Preliminary XFEL data from spontaneously grown endo-1,4-β-xylanase crystals from Hypocrea virens.

机构信息

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.

出版信息

Acta Crystallogr F Struct Biol Commun. 2022 Jun 1;78(Pt 6):226-231. doi: 10.1107/S2053230X22005118. Epub 2022 May 27.

Abstract

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.

摘要

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。

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