β-N-Acetylhexosaminidase-catalysed synthesis of non-reducing oligosaccharides

Rauvolfová, Jana; Kuzma, Marek; Weignerová, Lenka; Fialová, Pavla; Přikrylová, Věra; Pišvejcová, Andrea; Macková, Martina; Křen, Vladimı́r

doi:10.1016/j.molcatb.2003.10.008

Cited by 18 publications

(17 citation statements)

References 29 publications

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“…The properties of the CH11 enzyme were similar to those of Serratia plymuthica HRO‐C48, which has a molecular weight of 95.6 kDa and an optimal pH and temperature of 6.6 and 43 °C, respectively (Frankowski et al., ). In contrast, many β‐N‐acetylhexosaminidases from fungi showed maximum activity at an acidic pH around 5 (Rauvolfová et al., ; Weignerová et al., ).…”

Section: Resultsmentioning

confidence: 99%

“…For the transglycosylation reaction, it is likely that the pH shift is related to the structural changes of the active site as it makes space for the acceptor. Incidentally, the derivatives with a p NP group, such as p NP‐GlcNAc and p NP‐GalNAc, are used as donors by β‐N‐acetylhexosaminidases from many species (Chen et al., ; Ogata et al., ; Rauvolfová et al., ; Singh et al., ). Chitin oligosaccharides such as N, N′‐diacetylchitobiose are available from chitin easily and cheaply by acid hydrolysis.…”

Section: Resultsmentioning

confidence: 99%

“…Serratia mascescens YS‐1 enzyme also showed high specificity for the dialcohols 1,4‐butanediol, 1,5‐pentanediol and 1,6‐hexanediol, but was less specific toward xylitol. β‐N‐Acetylhexosaminidases from Bifidobacterium bifidum , Aspergillus flavofurcatis , and Aspergillus oryzae were shown to catalyze transglycosylation reactions with lactose, galactose and N‐acetylglucosamine as acceptors in contrast to the action of the CH11 enzyme (Chen et al., ; Rauvolfová et al., ; Singh et al., ).…”

Section: Resultsmentioning

confidence: 99%

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Characteristics of an β‐N‐Acetylhexosaminidase from Bacillus sp. CH11, Including its Transglycosylation Activity

Kurakake

Amai

Konishi

et al. 2018

Journal of Food Science

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The new transfer products including 1-O-β-D-N-acetylglucosaminyl xylitol are attractive compounds for their potential physiological functions. 1-O-β-D-N-Acetylglucosaminyl xylitol was produced effectively from chitin-oligosaccharides and xylitol by β-N-acetylhexosaminidase from Bacillus sp. CH11. This enzyme may be useful for the development of food materials for health-related applications such as oligosaccharides with intestinal functions and noncariogenic sugars.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Characteristics of an β‐N‐Acetylhexosaminidase from Bacillus sp. CH11, Including its Transglycosylation Activity

Kurakake

Amai

Konishi

et al. 2018

Journal of Food Science

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“…The reaction mixture, containing 1% colloidal chitin and 0.5 U of purified chitinase (Echi47), was incubated at 40°C for 2 h. Hydrolysis products were analyzed by thin-layer chromatography (TLC). 25 Protease Treatments. To determine resistance to proteases, the purified Echi47 (4 mU) was incubated with pepsin (pH 2.0; Sigma), trypsin (pH 7.0; Sigma), proteinase K (pH 7.5; Sigma), and flavor protease (pH 7.0; Novozymes) at a ratio of 1:10 (Echi47/protease, w/ w) at 37°C for 1 h. The residual activities were determined using the standard assay.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Novel Protease-Resistant Exochitinase (Echi47) from Pig Fecal Environment DNA with Application Potentials in the Food and Feed Industries

Liu

Yan

Yang

et al. 2015

J. Agric. Food Chem.

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A novel exochitinase gene (Echi47) was directly cloned from the pig fecal environment DNA using the genomic walking PCR technique and expressed in Escherichia coli BL21 (DE3). Echi47 has an open reading frame (ORF) of 1,161 bp encoding 386 amino acids. The amino acid sequence of Echi47 showed 36% identity with that of chitinase from Coprinellus congregatus. The recombinant exochitinase was purified with specific activity toward colloidal chitin of 6.84 U/mg. Echi47 was optimally active at pH 5.0 and 40 °C, respectively. When colloidal chitin was used as substrate, N-acetylchitobiose [(GlcNAc)2] was mostly produced at the initial stage, suggesting that it is an exochitinase. Echi47 exhibited excellent resistance to pepsin, trypsin, proteinase K, and flavor protease. Under simulated alimentary tract conditions, Echi47 was stable and active, releasing 21.1 mg of N-acetylchitooligosaccharides from 80 mg of colloidal chitin. These properties make Echi47 a potential additive in the food and feed industries.

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“…However, the available enzyme sources for synthesis remain limited. Most of the reported ␤-N-acetylhexosaminidases with transglycosylation ability are obtained from fungi, especially those belonging to the genera Aspergillus (22)(23)(24)(26)(27)(28)(29), Penicillium (28,30), and Talaromyces (31,32), and only three are obtained from the bacteria Nocardia orientalis and Serratia marcescens YS-1 (33)(34)(35). Moreover, most ␤-N-acetylhexosaminidases with transglycosylation ability display low glycosyl transfer efficiency at 1% to 10% yield, as well as poor regioselectivity, resulting in isomer production with multiple glycosyl linkages that are difficult to isolate (33,35,36).…”

Section: N-acetyl-␤-d-hexosamine (Hexnac) N-acetyl-␤-d-galactosaminementioning

confidence: 99%

Efficient and Regioselective Synthesis of β-GalNAc/GlcNAc-Lactose by a Bifunctional Transglycosylating β- N -Acetylhexosaminidase from Bifidobacterium bifidum

Chen

Jin

et al. 2016

Appl Environ Microbiol

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ABSTRACT␤-N-Acetylhexosaminidases have attracted interest particularly for oligosaccharide synthesis, but their use remains limited by the rarity of enzyme sources, low efficiency, and relaxed regioselectivity of transglycosylation. In this work, genes of 13 ␤-Nacetylhexosaminidases, including 5 from Bacteroides fragilis ATCC 25285, 5 from Clostridium perfringens ATCC 13124, and 3 from Bifidobacterium bifidum JCM 1254, were cloned and heterogeneously expressed in Escherichia coli. The resulting recombinant enzymes were purified and screened for transglycosylation activity. A ␤-N-acetylhexosaminidase named BbhI, which belongs to glycoside hydrolase family 20 and was obtained from B. bifidum JCM 1254, possesses the bifunctional property of efficiently transferring both GalNAc and GlcNAc residues through ␤1-3 linkage to the Gal residue of lactose. The effects of initial substrate concentration, pH, temperature, and reaction time on transglycosylation activities of BbhI were studied in detail. With the use of 10 mM pNP-␤-GalNAc or 20 mM pNP-␤-GlcNAc as the donor and 400 mM lactose as the acceptor in phosphate buffer (pH 5.8), BbhI synthesized GalNAc␤1-3Gal␤1-4Glc and GlcNAc␤1-3Gal␤1-4Glc at maximal yields of 55.4% at 45°C and 4 h and 44.9% at 55°C and 1.5 h, respectively. The model docking of BbhI with lactose showed the possible molecular basis of strict regioselectivity of ␤1-3 linkage in ␤-N-acetylhexosaminyl lactose synthesis. IMPORTANCEOligosaccharides play a crucial role in many biological events and therefore are promising potential therapeutic agents. However, their use is limited because large-scale production of oligosaccharides is difficult. The chemical synthesis requires multiple protecting group manipulations to control the regio-and stereoselectivity of glycosidic bonds. In comparison, enzymatic synthesis can produce oligosaccharides in one step by using glycosyltransferases and glycosidases. Given the lower price of their glycosyl donor and their broader acceptor specificity, glycosidases are more advantageous than glycosyltransferases for large-scale synthesis. ␤-N-Acetylhexosaminidases have attracted interest particularly for ␤-N-acetylhexosaminyl oligosaccharide synthesis, but their application is affected by having few enzyme sources, low efficiency, and relaxed regioselectivity of transglycosylation. In this work, we describe a microbial ␤-N-acetylhexosaminidase that exhibited strong transglycosylation activity and strict regioselectivity for ␤-N-acetylhexosaminyl lactose synthesis and thus provides a powerful synthetic tool to obtain biologically important GalNAc␤1-3Lac and GlcNAc␤1-3Lac. Oligosaccharides are widely distributed in nature and play a crucial role in many biological events, such as cell structure modulation, cell-cell recognition and communication, and cellmicrobe/toxin interaction and adhesion; therefore, they are promising and important potential therapeutic agents (1-3). However, their use is limited because of the difficulty of large-scale production of oligosaccharides. Their...

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β-N-Acetylhexosaminidase-catalysed synthesis of non-reducing oligosaccharides

Cited by 18 publications

References 29 publications

Characteristics of an β‐N‐Acetylhexosaminidase from Bacillus sp. CH11, Including its Transglycosylation Activity

Characteristics of an β‐N‐Acetylhexosaminidase from Bacillus sp. CH11, Including its Transglycosylation Activity

Novel Protease-Resistant Exochitinase (Echi47) from Pig Fecal Environment DNA with Application Potentials in the Food and Feed Industries

Efficient and Regioselective Synthesis of β-GalNAc/GlcNAc-Lactose by a Bifunctional Transglycosylating β- N -Acetylhexosaminidase from Bifidobacterium bifidum

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