Determination of the Catalytic Base in Family 48 Glycosyl Hydrolases

Maxim Kostylev; David B. Wilson

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Determination of the Catalytic Base in Family 48 Glycosyl Hydrolases

机译：48族糖基水解酶中催化碱的测定

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The catalytic base in family 48 glycosyl hydrolases has not been previously established experimentally. Based on structural and modeling data published to date, we used site-directed mutagenesis and azide rescue activity assays to show definitively that the catalytic base in Thermobifida fusca Cel48A is aspartic acid 225. Of the tested mutants, only Cel48A with the D225E mutation retained partial activity on soluble and insoluble substrates. In azide rescue experiments, only the D225G mutation, in the smallest residue tested, showed an increase in activity with added azide.

机译：家族48糖基水解酶的催化碱基以前尚未通过实验建立。根据迄今发布的结构和模型数据，我们使用了定点诱变和叠氮化物拯救活性分析法，明确显示了Thermobifida fusca Cel48A的催化碱基是天冬氨酸225。在测试的突变体中，只有具有D225E突变的Cel48A保留了部分对可溶和不可溶底物的活性。在叠氮化物拯救实验中，只有D225G突变（在最小的残基测试中）显示出添加叠氮化物后活性增加。

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《Applied Microbiology》 |2011年第17期|共3页
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Maxim Kostylev; David B. Wilson;
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1. Properties and gene structure of a bifunctional cellulolytic enzyme (CelA) from the extreme thermophile ‘Anaerocellum thermophilum’ with separate glycosyl hydrolase family 9 and 48 catalytic domains [J] . Vladimir Zverlov, Sabine Mahr, Kathrin Riedel, Microbiology . 1998,第2期

机译：用单独的糖基水解酶9和48催化结构域的极端嗜热纤维化酶（CELA）的双官能纤维素分解酶（CELA）的性质和基因结构
2. Nucleotide sequence of arfB of Clostridium stercorarium, and prediction of catalytic residues of alpha L-arabinofuranosidases based on local similarity with several families of glycosyl hydrolases [J] . Zverlov VV., Bachleitner M., Schwarz WH., FEMS Microbiology Letters . 1998,第2期

机译：梭状芽孢杆菌arfB的核苷酸序列，以及基于与糖基水解酶几个家族的局部相似性预测αL-阿拉伯呋喃糖苷酶的催化残基
3. Diversity of Bacteria and Glycosyl Hydrolase Family 48 Genes in Cellulolytic Consortia Enriched from Thermophilic Biocompost [J] . Javier A. Izquierdo, Maria V. Sizova, Lee R. Lynd Applied and Environmental Microbiology . 2010,第11期

机译：嗜热生物堆富集的纤维素分解财团中细菌和糖基水解酶家族48基因的多样性。
4. VARIANTS OF GLYCOSYL HYDROLASE FAMILY 2 P-GLUCURONIDASES [C] . Caleb R. Schlachter, Pongkwan N. Sitasuwan, Amanda C. McGee, Conference on biochemical and molecular engineering . 2019

机译：各种糖基水解酶家族2 P-葡糖醛酸苷酶
5. Endoxylanases from glycosyl hydrolase families 5 and 10: Bacterial enzymes for development of gram -positive biocatalysts for production of bio -based products. [D] . St. John, Franz Josef. 2006

机译：来自糖基水解酶家族5和10的内切木聚糖酶：用于开发革兰氏阳性生物催化剂以生产生物基产品的细菌酶。
6. Determination of the Catalytic Base in Family 48 Glycosyl Hydrolases [O] . Maxim Kostylev, David B. Wilson 2011

机译：48种糖基水解酶中催化碱的测定
7. Determination of the Catalytic Base in Family 48 Glycosyl Hydrolases ▿ [O] . Kostylev, Maxim, Wilson, David B. 2011

机译：家族48个糖基水解酶中催化碱的测定▿

Determination of the Catalytic Base in Family 48 Glycosyl Hydrolases

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