Biocatalytic Synthesis of D-Allulose Using Novel D-Tagatose 3-Epimerase From Christensenella minuta

Abstract

D-allulose, which is one of the important rare sugars, has gained significant attention in the food and pharmaceutical industries as a potential alternative to sucrose and fructose. Enzymes belonging to the D-tagatose 3-epimerase (DTEase) family can reversibly catalyze the epimerization of D-fructose at the C3 position and convert it into D-allulose by a good number of naturally occurring microorganisms. However, microbial synthesis of D-allulose is still at its immature stage in the industrial arena, mostly due to the preference of slightly acidic conditions for Izumoring reactions. Discovery of novel DTEase that works at acidic conditions is highly preferred for industrial applications. In this study, a novel DTEase, DTE-CM, capable of catalyzing D-fructose into D-allulose was applications. In this study, a novel DTEase, DTE-CM, capable of catalyzing D-fructose into D-allulose was DTE-CM on D-fructose was found to be remarkably influenced and modulated by the type of metal ions (co-factors). The DTE-CM on D-fructose was found to be remarkably influenced and modulated by the type of metal ions (co-factors). The 50°C from 0.5 to 3.5 h at a concentration of 0.1 mM. The enzyme exhibited its maximum catalytic activity on D-fructose at pH 6.0 and 50°C from 0.5 to 3.5 h at a concentration of 0.1 mM. The enzyme exhibited its maximum catalytic activity on -fructose at pH 6.0 and 50°C with a Kcat/Km value of 45 mM−1min−1. The 500 g/L D-fructose, which corresponded to 30% conversion rate. With these interesting catalytic properties, this enzyme could be a promising candidate for industrial biocatalytic applications.

Crystal structure of a novel homodimericD-allulose 3-epimerase from a Clostridia bacterium

“…1). The polygenetic tree of the KEase family obtained by the neighbor-joining method (Wang et al, 2021) indicated a close relationship of CDAE to other DAEases; in particular, an evolutionary relationship between CDAE and the DAEases from Clostridium cellulolyticum H10 (Clce-DAE; GenBank ACL75304.1; Mu et al, 2011) and Clostridium sp. BNL1100 (Clsp-DAE; GenBank WP_014314767.1; Mu et al, 2013).…”

Research Advances of d-allulose: An Overview of Physiological Functions, Enzymatic Biotransformation Technologies, and Production Processes

“…Notably, only the DPEase from Dorea sp. CAG317 and the DPEase from C. minuta are optimally adapted to weak acidic pH conditions [24,25], suggesting that these two enzymes have potential for application in the large-scale production of D-allulose. Note: NR = not reported.…”

“…Most of them are weakly basophilic [ 22 , 23 ], except for Dorea sp. CAG317 d -psicose 3-epimerase (DPEase) and Christensenella minuta DPEase, which posess a weakly acidic optimal pH [ 24 , 25 ]. Ketose 3-epimerases have been heterologously expressed successfully in prokaryotic and eukaryotic systems for the efficient production of D-allulose and its high-value derivates.…”

The Characterization of a Novel D-allulose 3-Epimerase from Blautia produca and Its Application in D-allulose Production

“…The Izumoring strategy is an effective method proposed in 2006 for the bioproduction of various kinds of rare hexoses [ 18 ] and also involves D-allulose 3-epimerase (DAE), D-tagatose 3-epimerase (DTE), polyol dehydrogenases, and aldose isomerases [ 19 ]. Consequently, D-fructose can be epimerized at the C-3 position to produce D-allulose by DTEs or DAEs [ 20 ]. Due to the substrate specificity, catalytic property, and specific enzyme activity of D-tagatose epimerase and D-allulose epimerase being different enzymes and are named DTE and DAE, respectively [ 21 , 22 ].…”

“…According to one study, DTE extracted from the genes found in the macro-genome resource of the hot water exhibited high thermostability at 60 °C to 70 °C [ 35 ]. Probiotics are also sources of several DTEs, including the DTE-CM gene from Christensenella minuta DSM 22607 [ 20 ], which is produced in accordance with food safety regulations.…”