Mapping the Transglycosylation Relevant Sites of Cold-Adapted β-D-Galactosidase from Arthrobacter sp. 32cB - Publikacja - MOST Wiedzy

Wyszukiwarka

Mapping the Transglycosylation Relevant Sites of Cold-Adapted β-D-Galactosidase from Arthrobacter sp. 32cB

Abstrakt

B-Galactosidase from Arthrobacter sp. 32cB (ArthbetaDG) is a cold-adapted enzyme able to catalyze hydrolysis of beta-D-galactosides and transglycosylation reaction, where galactosyl moiety is being transferred onto an acceptor larger than a water molecule. Mutants of ArthbetaDG D207A and E517Q were designed to determine the significance of specific residues and to enable formation of complexes with lactulose and sucrose and to shed light onto the structural basis of the transglycosylation reaction. The catalytic assays proved loss of function mutation E517 into glutamine and a significant drop of activity for mutation of D207 into alanine. Solving crystal structures of two new mutants, and new complex structures of previously presented mutant E441Q enables description of introduced changes within active site of enzyme and determining the importance of mutated residues for active site size and character. Furthermore, usage of mutants with diminished and abolished enzymatic activity enabled solving six complex structures with galactose, lactulose or sucrose bounds. As a result, not only the galactose binding sites were mapped on the enzyme’s surface but also the mode of lactulose, product of transglycosylation reaction, and binding within the enzyme’s active site were determined and the glucopyranose binding site in the distal of active site was discovered. The latter two especially show structural details of transglycosylation, providing valuable information that may be used for engineering of ArthbetaDG or other analogous galactosidases belonging to GH2 family.

Cytowania

  • 3

    CrossRef

  • 2

    Web of Science

  • 3

    Scopus

Autorzy (3)

Cytuj jako

Pełna treść

pobierz publikację
pobrano 8 razy
Wersja publikacji
Accepted albo Published Version
Licencja
Creative Commons: CC-BY otwiera się w nowej karcie

Słowa kluczowe

Informacje szczegółowe

Kategoria:
Publikacja w czasopiśmie
Typ:
artykuły w czasopismach
Opublikowano w:
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES nr 21,
ISSN: 1661-6596
Język:
angielski
Rok wydania:
2020
Opis bibliograficzny:
Rutkiewicz M., Wanarska M., Bujacz A.: Mapping the Transglycosylation Relevant Sites of Cold-Adapted β-D-Galactosidase from Arthrobacter sp. 32cB// INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES -Vol. 21,iss. 15 (2020), s.5354-
DOI:
Cyfrowy identyfikator dokumentu elektronicznego (otwiera się w nowej karcie) 10.3390/ijms21155354
Weryfikacja:
Politechnika Gdańska

wyświetlono 5 razy

Publikacje, które mogą cię zainteresować

Meta Tagi