Characterization of a new α-l-fucosidase isolated from Fusarium proliferatum LE1 that is regioselective to α-(1 → 4)-l-fucosidic linkage in the hydrolysis of α-l-fucobiosides

Characterization of a new α-l-fucosidase isolated from Fusarium proliferatum LE1 that is regioselective to α-(1 → 4)-l-fucosidic linkage in the hydrolysis of α-l-fucobiosides Научная публикация

Журнал

Biochimie
ISSN: 1638-6183 , E-ISSN: 6183-1638

Вых. Данные

Год: 2017, Том: 132, Страницы: 54-65 Страниц : 12 DOI: 10.1016/j.biochi.2016.10.014

Авторы

Shvetsova Svetlana V. ^1,2 , Shabalin Konstantin A. ^2,3 , Bobrov Kirill S. ² , Ivanen Dina R. ² , Ustyuzhanina Nadezhda E. ⁴ , Krylov Vadim B. ⁴ , Nifantiev Nikolay E. ⁴ , Naryzhny Stanislav N. ^5,2 , Zgoda Victor G. ⁵ , Eneyskaya Elena V. ² , Kulminskaya Anna A. ^1,2

Организации

1	Department of Medical Physics, Peter the Great St. Petersburg Polytechnic University, 195251, St. Petersburg, Chlopina Str. 5, Russia
2	National Research Center «Kurchatov Institute», B.P. Konstantinov Petersburg Nuclear Physics Institute, 188300, Gatchina, Orlova Roscha, Russia
3	Peter the Great St. Petersburg Polytechnic University, 195251, St. Petersburg, Polytechnicheskaya Str., 29, Russia
4	N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 47, Leninsky Pr., Moscow, 119991, Russia
5	Institute of Biomedical Chemistry, Pogodinskaya 10, Moscow, 119121, Russia

Here, we report the biochemical characterization of a novel α-l-fucosidase with broad substrate specificity (FpFucA) isolated from the mycelial fungus Fusarium proliferatum LE1. Highly purified α-l-fucosidase was obtained from several chromatographic steps after growth in the presence of l-fucose. The purified α-l-fucosidase appeared to be a monomeric protein of 67 ± 1 kDa that was able to hydrolyze the synthetic substrate p-nitrophenyl α-l-fucopyranoside (pNPFuc), with Km = 1.1 ± 0.1 mM and kcat = 39.8 ± 1.8 s−1. l-fucose, 1-deoxyfuconojirimycin and tris(hydroxymethyl)aminomethane inhibited pNPFuc hydrolysis, with inhibition constants of 0.2 ± 0.05 mM, 7.1 ± 0.05 nM, and 12.2 ± 0.1 mM, respectively. We assumed that the enzyme belongs to subfamily A of the GH29 family (CAZy database) based on its ability to hydrolyze practically all fucose-containing oligosaccharides used in the study and the phylogenetic analysis. We found that this enzyme was a unique α-l-fucosidase that preferentially hydrolyzes the α-(1 → 4)-L-fucosidic linkage present in α-L-fucobiosides with different types of linkages. As a retaining glycosidase, FpFucA is capable of catalyzing the transglycosylation reaction with alcohols (methanol, ethanol, and 1-propanol) and pNP-containing monosaccharides as acceptors. These features make the enzyme an important tool that can be used in the various modifications of valuable fucose-containing compounds.

Shvetsova S.V. , Shabalin K.A. , Bobrov K.S. , Ivanen D.R. , Ustyuzhanina N.E. , Krylov V.B. , Nifantiev N.E. , Naryzhny S.N. , Zgoda V.G. , Eneyskaya E.V. , Kulminskaya A.A.
Characterization of a new α-l-fucosidase isolated from Fusarium proliferatum LE1 that is regioselective to α-(1 → 4)-l-fucosidic linkage in the hydrolysis of α-l-fucobiosides
Biochimie. 2017. V.132. P.54-65. DOI: 10.1016/j.biochi.2016.10.014 WOS Scopus OpenAlex

≡ Web of science:	WOS:000392687300007
≡ Scopus:	2-s2.0-84995513642
≡ OpenAlex:	W2542664455