Crystal structure of a glycoside hydrolase family 68 β-fructosyltransferase from Beijerinckia indica subsp. indica in complex with fructose

Crystal structure of a glycoside hydrolase family 68 β-fructosyltransferase from Beijerinckia indica subsp. indica in complex with fructose
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Beijerinckia indica indica 糖苷水解酶家族 68 β-果糖基转移酶与果糖复合物的晶体结构。

DOI:
10.1080/09168451.2020.1804317
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发表时间:
2020
期刊:
Biosci. Biotechnol. Biochem.
影响因子:
--
通讯作者:
T.
T.
中科院分区:
--
文献类型:
--
作者:
Tonozuka;T.;Kitamura;J.;Nagaya;M.;Kawai;R.;Nishikawa;A.;Katsuaki Hirano;K.;Tamura;K.;Fujii;T. and Tochio;T.

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在大肠杆菌中表达了一种属于Beijerinckia indicasubsp.indicaNBRC 3744糖苷水解酶家族68(GH 68)的酶。酶的生化特性分析表明,该酶是一种β-果糖基转移酶(BiBftA)。最初尝试了全长BiBfTA的结晶,但没有获得晶体。我们构建了一个变体,其中在潜在的柔性区域中的5个残基(Pro 199-Gly 203)和13个残基(Leu 522-Gln 534)被缺失,并且我们成功地使该变体BiBftA结晶。BiBftA与其他GH 68酶一样由五叶β螺旋桨折叠组成。与果糖复合的BiBftA的结构出乎意料地表明,一个β-呋喃果糖(β-Fruf)分子和一个β-吡喃果糖分子与催化口袋结合。β-Frufat亚位点-1的取向与大多数GH 68酶中观察到的取向倾斜,呈现出GH 68酶的第二种结构与β-Fruf的倾斜结合模式复合。
An enzyme belonging to glycoside hydrolase family 68 (GH68) fromBeijerinckia indicasubsp.indicaNBRC 3744 was expressed inEscherichia coli. Biochemical characterization showed that the enzyme was identified to be a β-fructosyltransferase (BiBftA). Crystallization of a full-length BiBftA was initially attempted, but no crystals were obtained. We constructed a variant in which 5 residues (Pro199-Gly203) and 13 residues (Leu522-Gln534) in potentially flexible regions were deleted, and we successfully crystallized this variant BiBftA. BiBftA is composed of a five-bladed β-propeller fold as in other GH68 enzymes. The structure of BiBftA in complex with fructose unexpectedly indicated that one β-fructofuranose (β-Fruf) molecule and one β-fructopyranose molecule bind to the catalytic pocket. The orientation of β-Frufat subsite −1 is tilted from the orientation observed in most GH68 enzymes, presenting a second structure of a GH68 enzyme in complex with the tilted binding mode of β-Fruf.
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