Enzymatic Synthesis of Maltitol and Its Inhibitory Effect on the Growth of Streptococcus mutans DMST 18777.

Patinya Haewpetch,Jarunee Kaulpiboon,Prakarn Rudeekulthamrong

doi:10.3390/biom12020167

Patinya Haewpetch, Jarunee Kaulpiboon + Show 1 more

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https://doi.org/10.3390/biom12020167

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Journal: Biomolecules	Publication Date: Jan 20, 2022
Citations: 2	License type: CC BY 4.0

Affiliation: Thammasat University, Phramongkutklao Hospital

Abstract

This study aimed to synthesize maltitol using recombinant CGTase from Bacillus circulans A11 with β-cyclodextrin (β-CD) and sorbitol as a glucosyl donor and acceptor, respectively, and assess its antibacterial activity. Optimal conditions for producing the highest yield, 25.0% (w/w), were incubation of 1% (w/v) β-CD and sorbitol with 400 U/mL of CGTase in 20 mM phosphate buffer at pH 6.0 and 50 °C for 72 h. Subsequently, maltitol underwent large-scale production and was purified by HPLC. By mass spectrometry, the molecular weight of the synthesized maltitol was 379.08 daltons, corresponding exactly to that of standard maltitol. The relative sweetness of synthesized and standard maltitol was ~90% of that of sucrose. Spot assay on the agar plate showed that maltitol inhibited the growth of Streptococcus mutans DMST 18777 cells. In addition, the MIC and MBC values of synthesized and standard maltitol against S. mutans were also determined as 20 and 40 mg/mL, respectively. These results show that the synthesized maltitol can be produced at high yields and has the potential to be used as an anticariogenic agent in products such as toothpaste.

Highlights

Enzymatic transglycosylation has been utilized in food and medicinal chemistry to synthesize oligosaccharides, polysaccharides and saccharide derivatives, which can lead to changes in their physiological and chemical properties such as water solubility and sensory characteristics such as mouth-feel taste [1,2]
The results showed that maltose had the highest activity, so it was set as 100% relative activity, followed by maltitol (C12, 4-O-α-glucopyranosyl-D-sorbitol), sorbitol (C6 ; six-carbon sugar alcohol), mannitol (C6 ), galactitol (C6 ), xylitol (C5 ), arabitol (C5 ), and erythritol (C4 ), at 48.7, 45.6, 44.2, 42.4, 41.6, 35.1 and 33.9% of activity, respectively (Table 1)
These results suggested that cyclodextrin glycosyltransferase (CGTase) preferred the disaccharide-polyol acceptor to monosaccharide-polyol acceptors, given the high coupling activity on maltose and maltitol

Summary

Introduction

Enzymatic transglycosylation has been utilized in food and medicinal chemistry to synthesize oligosaccharides, polysaccharides and saccharide derivatives, which can lead to changes in their physiological and chemical properties such as water solubility and sensory characteristics such as mouth-feel taste (sweetness, bitterness and astringency) [1,2]. Cyclodextrin glycosyltransferase (CGTase—E.C. 2.4.1.19), a glycosidase, is an enzyme that catalyzes the transglycosylation reaction, which transfers the α-1,4 glucan from a glycosyl donor to another glucan acceptor such as starch, related oligo/polysaccharides, and sugar alcohols with a free 4-hydroxyl group [3]. The transglycosylation of CGTase consists of intramolecular and intermolecular transglycosylation reactions. Intramolecular transglycosylation (cyclization reaction) results in the formation of cyclic oligosaccharides, e.g., cyclodextrin (CD) and intermolecular transglycosylation can occur through three different reactions. The major reaction is disproportionation, in which a glycosyl is transferred from a donor to an acceptor oligo/polysaccharide or sugar alcohol with a free 4-hydroxyl group resulting in a linear oligo/polysaccharide or sugar alcohol of variable length. The second reaction is coupling, a reversal of cyclization, and the third, hydrolysis, is a low activity reaction with water acting as a glycosyl acceptor [4,5,6,7,8]

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