Identification of difructose dianhydride I synthase/hydrolase from an oral bacterium establishes a novel glycoside hydrolase family.
Identification of difructose dianhydride I synthase/hydrolase from an oral bacterium establishes a novel glycoside hydrolase family.
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DOI：
10.1016/j.jbc.2021.101324
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发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Fushinobu S
中科院分区：
文献类型：
--
作者：
Kashima T;Okumura K;Ishiwata A;Kaieda M;Terada T;Arakawa T;Yamada C;Shimizu K;Tanaka K;Kitaoka M;Ito Y;Fujita K;Fushinobu S
低聚果糖及其酸酐被广泛用作促进健康的食品和益生元。各种酶作用于天然果聚糖聚合物的β- d -构呋喃基键，已被用于生产功能化合物。然而，对水解和形成α- d -构呋喃基键的酶的研究较少。本研究鉴定了来自牙双歧杆菌的BBDE_2040基因产物（α- d -果糖呋喃糖苷酶和来自牙双歧杆菌的二果糖二酐I合成酶/水解酶[αFFase1]）是一种具有α- d -果糖呋喃糖苷酶和α- d -阿拉伯糖呋喃糖苷酶活性的酶，并具有保持异构体的方式。αFFase1与任何已知的酶都不同源，这表明它是一个新的糖苷水解酶家族的成员。将焦糖果糖与αFFase1孵育后，观察到β-D-Fruf-(2→1)-α- d - fruf -1,2′：2,1′-β-D-Fruf（二异戊糖II）和β-D-Fruf-(2→1)-α- d - fruf（二糖糖）到α- d - fruf -1,2′：2,1′-β-D-Fruf（二果糖二酐I [DFA I]）的转化。菊糖糖与DFA I的反应平衡偏向后者（1:9），促进了分子内脱水缩合反应。因此，我们将这种酶命名为DFA I合成酶/水解酶。α - ffase1与β-D-Fruf和β-D-Araf配合物的晶体结构以高达1.76 Å的分辨率测定。DFA I分子在活性位点的建模和突变分析也确定了催化和底物结合的关键残基。α - ffase1的六聚体结构揭示了催化口袋通过通道连接到一个大的内部腔。分子动力学分析表明DFA I和菊糖与周围水分子稳定结合在活性位点。综上所述，这些结果确定了DFA I合成酶/水解酶是一个新的糖苷水解酶家族（GH172）的成员。
Fructooligosaccharides and their anhydrides are widely used as health-promoting foods and prebiotics. Various enzymes acting on β-D-fructofuranosyl linkages of natural fructan polymers have been used to produce functional compounds. However, enzymes that hydrolyze and form α-D-fructofuranosyl linkages have been less studied. Here, we identified the BBDE_2040 gene product from Bifidobacterium dentium (α-D-fructofuranosidase and difructose dianhydride I synthase/hydrolase from Bifidobacterium dentium [αFFase1]) as an enzyme with α-D-fructofuranosidase and α-D-arabinofuranosidase activities and an anomer-retaining manner. αFFase1 is not homologous with any known enzymes, suggesting that it is a member of a novel glycoside hydrolase family. When caramelized fructose sugar was incubated with αFFase1, conversions of β-D-Frup-(2→1)-α-D-Fruf to α-D-Fruf-1,2′:2,1′-β-D-Frup (diheterolevulosan II) and β-D-Fruf-(2→1)-α-D-Fruf (inulobiose) to α-D-Fruf-1,2′:2,1′-β-D-Fruf (difructose dianhydride I [DFA I]) were observed. The reaction equilibrium between inulobiose and DFA I was biased toward the latter (1:9) to promote the intramolecular dehydrating condensation reaction. Thus, we named this enzyme DFA I synthase/hydrolase. The crystal structures of αFFase1 in complex with β-D-Fruf and β-D-Araf were determined at the resolutions of up to 1.76 Å. Modeling of a DFA I molecule in the active site and mutational analysis also identified critical residues for catalysis and substrate binding. The hexameric structure of αFFase1 revealed the connection of the catalytic pocket to a large internal cavity via a channel. Molecular dynamics analysis implied stable binding of DFA I and inulobiose to the active site with surrounding water molecules. Taken together, these results establish DFA I synthase/hydrolase as a member of a new glycoside hydrolase family (GH172).
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影响因子：
5.5
作者：
Hess, Berk;Kutzner, Carsten;Lindahl, Erik
通讯作者：
Lindahl, Erik
使用 (1) H NMR 光谱观察 D(2)O 中 D-果糖的酮互变异构体。
影响因子：
3.1
作者：
Barclay, Thomas;Ginic-Markovic, Milena;Johnston, Martin R.;Cooper, Peter;Petrovsky, Nikolai
通讯作者：
Petrovsky, Nikolai
影响因子：
5.5
作者：
Hess, Berk
通讯作者：
Hess, Berk
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
4.4
作者：
ESSMANN, U;PERERA, L;PEDERSEN, LG
通讯作者：
PEDERSEN, LG