The variable structural flexibility of the Bacillus circulans
β-galactosidase isoforms determines their unique
functionalities

J 2024

The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities

HOVORKOVA, Michaela; Barbora KASCAKOVA; Lucie PETRASKOVA; Petra HAVLICKOVA; Jiří NOVÁČEK et. al.

Základní údaje

Originální název

The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities

Autoři

HOVORKOVA, Michaela; Barbora KASCAKOVA; Lucie PETRASKOVA; Petra HAVLICKOVA; Jiří NOVÁČEK; Daniel PINKAS; Zdenko GARDIAN; Vladimir KREN; Pavla BOJAROVA a Ivana Kuta SMATANOVA

Vydání

Structure, CAMBRIDGE, CELL PRESS, 2024, 0969-2126

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.300

Kód RIV

RIV/00216224:14740/24:00139160

Organizační jednotka

Středoevropský technologický institut

DOI

https://doi.org/10.1016/j.str.2024.09.005

UT WoS

001354882000001

EID Scopus

2-s2.0-85207765085

Klíčová slova anglicky

CRYO-EMOLIGOSACCHARIDE PRODUCTION; REFINEMENT; MECHANISMS; CLONING; SYSTEM; MODEL; GENE

Štítky

CF CRYO, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 9. 6. 2025 15:52, Mgr. Eva Dubská

Anotace

V originále

beta-Galactosidase from Bacillus circulans ATCC 31382 (BgaD) is a biotechnologically important enzyme for the synthesis of beta-galactooligosaccharides (GOS). Among its four isoforms, isoform A (BgaD-A) has distinct synthetic properties. Here, we present cryoelectron microscopy (cryo-EM) structures of BgaD-A and compare them with the known X-ray crystal structure of isoform D (BgaD-D), revealing substantial structural divergences between the two isoforms. In contrast to BgaD-D, BgaD-A features a flexible Big-4 domain and another enigmatic domain. The newly identified flexible region in BgaD-A is termed as "barrier domain 8," and serves as a barricade, obstructing the access of longer oligosaccharide substrates into the active site of BgaD-A. The transgalactosylation reactions catalyzed by both isoforms revealed that BgaD-A has a higher selectivity than BgaD-D in the earlier stages of the reaction and is prevailingly directed to shorter galactooligosaccharides. This study improves our understanding of the structural determinants governing R-galactosidase catalysis, with implications for tailored GOS production.

Návaznosti

EF18_046/0015974, projekt VaV

Název: Modernizace České infrastruktury pro integrativní strukturní biologii

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

LM2023050, projekt VaV

Název: Národní infrastruktura pro biologické a medicínské zobrazování

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech BioImaging: Národní výzkumná infrastruktura pro biologické a medicínské zobrazování

Citovat

HOVORKOVA, Michaela; Barbora KASCAKOVA; Lucie PETRASKOVA; Petra HAVLICKOVA; Jiří NOVÁČEK; Daniel PINKAS; Zdenko GARDIAN; Vladimir KREN; Pavla BOJAROVA a Ivana Kuta SMATANOVA. The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities. Structure. CAMBRIDGE: CELL PRESS, 2024, roč. 32, č. 11, s. 2023-2037. ISSN 0969-2126. Dostupné z: https://doi.org/10.1016/j.str.2024.09.005.

@article{2486561,
   author = {Hovorkova, Michaela and Kascakova, Barbora and Petraskova, Lucie and Havlickova, Petra and Nováček, Jiří and Pinkas, Daniel and Gardian, Zdenko and Kren, Vladimir and Bojarova, Pavla and Smatanova, Ivana Kuta},
   article_location = {CAMBRIDGE},
   article_number = {11},
   doi = {https://doi.org/10.1016/j.str.2024.09.005},
   keywords = {CRYO-EMOLIGOSACCHARIDE PRODUCTION; REFINEMENT; MECHANISMS; CLONING; SYSTEM; MODEL; GENE},
   language = {eng},
   issn = {0969-2126},
   journal = {Structure},
   title = {The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities},
   url = {https://www.cell.com/structure/abstract/S0969-2126(24)00374-5?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS0969212624003745%3Fshowall%3Dtrue},
   volume = {32},
   year = {2024}
}

TY  - JOUR
ID  - 2486561
AU  - Hovorkova, Michaela - Kascakova, Barbora - Petraskova, Lucie - Havlickova, Petra - Nováček, Jiří - Pinkas, Daniel - Gardian, Zdenko - Kren, Vladimir - Bojarova, Pavla - Smatanova, Ivana Kuta
PY  - 2024
TI  - The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities
JF  - Structure
VL  - 32
IS  - 11
SP  - 2023-2037
EP  - 2023-2037
PB  - CELL PRESS
SN  - 09692126
KW  - CRYO-EMOLIGOSACCHARIDE PRODUCTION
KW  - REFINEMENT
KW  - MECHANISMS
KW  - CLONING
KW  - SYSTEM
KW  - MODEL
KW  - GENE
UR  - https://www.cell.com/structure/abstract/S0969-2126(24)00374-5?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS0969212624003745%3Fshowall%3Dtrue
N2  - beta-Galactosidase from Bacillus circulans ATCC 31382 (BgaD) is a biotechnologically important enzyme for the synthesis of beta-galactooligosaccharides (GOS). Among its four isoforms, isoform A (BgaD-A) has distinct synthetic properties. Here, we present cryoelectron microscopy (cryo-EM) structures of BgaD-A and compare them with the known X-ray crystal structure of isoform D (BgaD-D), revealing substantial structural divergences between the two isoforms. In contrast to BgaD-D, BgaD-A features a flexible Big-4 domain and another enigmatic domain. The newly identified flexible region in BgaD-A is termed as "barrier domain 8," and serves as a barricade, obstructing the access of longer oligosaccharide substrates into the active site of BgaD-A. The transgalactosylation reactions catalyzed by both isoforms revealed that BgaD-A has a higher selectivity than BgaD-D in the earlier stages of the reaction and is prevailingly directed to shorter galactooligosaccharides. This study improves our understanding of the structural determinants governing R-galactosidase catalysis, with implications for tailored GOS production.
ER  -

HOVORKOVA, Michaela; Barbora KASCAKOVA; Lucie PETRASKOVA; Petra HAVLICKOVA; Jiří NOVÁČEK; Daniel PINKAS; Zdenko GARDIAN; Vladimir KREN; Pavla BOJAROVA a Ivana Kuta SMATANOVA. The variable structural flexibility of the Bacillus circulans β-galactosidase isoforms determines their unique functionalities. \textit{Structure}. CAMBRIDGE: CELL PRESS, 2024, roč.~32, č.~11, s.~2023-2037. ISSN~0969-2126. Dostupné z: https://doi.org/10.1016/j.str.2024.09.005.

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