Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F22%3A00557244" target="_blank" >RIV/61388971:_____/22:00557244 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/22:10443847

Výsledek na webu

<a href="https://www.mdpi.com/1422-0067/23/8/4106" target="_blank" >https://www.mdpi.com/1422-0067/23/8/4106</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/ijms23084106" target="_blank" >10.3390/ijms23084106</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides

Popis výsledku v původním jazyce

Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is often complicated by the high price of sugar nucleotides, stringent substrate specificity, and low enzyme stability. Engineered glycosidases (EC 3.2.1) represent a good synthetic alternative, especially if variations in the substrate structure are desired. Site-directed mutagenesis can improve the synthetic process with higher yields and/or increased reaction selectivity. So far, the synthesis of human milk oligosaccharides by glycosidases has mostly been limited to analytical reactions with mass spectrometry detection. The present work reveals the potential of a library of engineered glycosidases in the preparative synthesis of three tetrasaccharides derived from lacto-N-tetraose (Gal beta 4GlcNAc beta 3Gal beta 4Glc), employing sequential cascade reactions catalyzed by beta 3-N-acetylhexosaminidase BbhI from Bifidobacterium bifidum, beta 4-galactosidase BgaD-B from Bacillus circulans, beta 4-N-acetylgalactosaminidase from Talaromyces flavus, and beta 3-galactosynthase BgaC from B. circulans. The reaction products were isolated and structurally characterized. This work expands the insight into the multi-step catalysis by glycosidases and shows the path to modified derivatives of complex carbohydrates that cannot be prepared by standard glycosyltransferase methods.

Název v anglickém jazyce

Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides

Popis výsledku anglicky

Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is often complicated by the high price of sugar nucleotides, stringent substrate specificity, and low enzyme stability. Engineered glycosidases (EC 3.2.1) represent a good synthetic alternative, especially if variations in the substrate structure are desired. Site-directed mutagenesis can improve the synthetic process with higher yields and/or increased reaction selectivity. So far, the synthesis of human milk oligosaccharides by glycosidases has mostly been limited to analytical reactions with mass spectrometry detection. The present work reveals the potential of a library of engineered glycosidases in the preparative synthesis of three tetrasaccharides derived from lacto-N-tetraose (Gal beta 4GlcNAc beta 3Gal beta 4Glc), employing sequential cascade reactions catalyzed by beta 3-N-acetylhexosaminidase BbhI from Bifidobacterium bifidum, beta 4-galactosidase BgaD-B from Bacillus circulans, beta 4-N-acetylgalactosaminidase from Talaromyces flavus, and beta 3-galactosynthase BgaC from B. circulans. The reaction products were isolated and structurally characterized. This work expands the insight into the multi-step catalysis by glycosidases and shows the path to modified derivatives of complex carbohydrates that cannot be prepared by standard glycosyltransferase methods.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2022

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

International Journal of Molecular Sciences

ISSN

1422-0067

e-ISSN

1422-0067

Svazek periodika

23

Číslo periodika v rámci svazku

8

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

13

Strana od-do

4106

Kód UT WoS článku

000786916300001

EID výsledku v databázi Scopus

2-s2.0-85127578440