Synthesis and characterization of stevioside having low degree polymerized glucosides using dextransucrase and dextranase

Son, Gyumin; Nguyen, Thi Thanh Hanh; Park, Byeongsu; Kwak, So-Hyung; Jin, Juhui; Kim, Young Min; Moon, Young Hwan; Park, Sunghee; Kim, Seong Bo; Kim, Doman

doi:10.1016/j.enzmictec.2019.109412

Cited by 13 publications

(5 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the other hand, glucansucrases are more promising enzymes in the modification of steviol glycosides, since they form a-1,6 and 1,3-glycosidic bonds, which are not hydrolyzed by human enzymes. Moreover, they can use low-cost donors, such as sucrose, reaching product yields over 90% (Ko et al, 2016;Devlamynck et al, 2019;Son et al, 2020). Regarding plantderived UDP-glucosyltransferase-mediated bioconversion, the regeneration of UDP sugar donors with sucrose synthase is necessary, but yields over 70% can be obtained (Wang et al, 2016;Chen et al, 2021Chen et al, , 2020Chen et al, , 2018.…”

Section: Discussionmentioning

confidence: 99%

β-Glucosidase and β-Galactosidase-Mediated Transglycosylation of Steviol Glycosides Utilizing Industrial Byproducts

Zerva

Chorozian

Kritikou

et al. 2021

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Stevia rebaudiana Bertoni is a plant cultivated worldwide due to its use as a sweetener. The sweet taste of stevia is attributed to its numerous steviol glycosides, however, their use is still limited, due to their bitter aftertaste. The transglycosylation of steviol glycosides, aiming at the improvement of their taste, has been reported for many enzymes, however, glycosyl hydrolases are not extensively studied in this respect. In the present study, a β-glucosidase, MtBgl3a, and a β-galactosidase, TtbGal1, have been applied in the transglycosylation of two steviol glycosides, stevioside and rebaudioside A. The maximum conversion yields were 34.6 and 33.1% for stevioside, while 25.6 and 37.6% were obtained for rebaudioside A conversion by MtBgl3a and TtbGal1, respectively. Low-cost industrial byproducts were employed as sugar donors, such as cellulose hydrolyzate and acid whey for TtbGal1- and MtBgl3a- mediated bioconversion, respectively. LC-HRMS analysis identified the formation of mono- and di- glycosylated products from stevioside and rebaudioside A. Overall, the results of the present work indicate that both biocatalysts can be exploited for the design of a cost-effective process for the modification of steviol glycosides.

show abstract

Section: Discussionmentioning

confidence: 99%

β-Glucosidase and β-Galactosidase-Mediated Transglycosylation of Steviol Glycosides Utilizing Industrial Byproducts

Zerva

Chorozian

Kritikou

et al. 2021

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…Stevioside (13-O-β-sophorosyl-19-O-β-D-glucosyl-steviol, Figure 8), the main component of Stevia leaves extract, was shown to be glucosylated by alternansucrase of Leuconostoc citreum SK24.002 with a conversion degree of over 70%, producing nine glucosyl-stevioside derivatives with up to three glucosyl units (Musa et al 2014). Under optimized conditions, dextransucrase of L. mesenteroides B-512 F/KM converted 98% of stevioside into its glucosides by transferring glucosyl units to the C-13-β-sophorosyl unit and/or the C-19-linked glucosyl unit with the formation of (α1→6) linkages (Son et al 2020). Gtf180-ΔN was also shown to be capable of glucosylating stevioside with the formation of multiple glucosides containing up to nine glucosyl units (Devlamynck et al 2019).…”

Section: Glucosylation Of Steviol Glycosides By Glucansucrasesmentioning

confidence: 99%

Enzymatic glucosylation of polyphenols using glucansucrases and branching sucrases of glycoside hydrolase family 70

Meng

Leeuw³

et al. 2021

Critical Reviews in Food Science and Nutrition

View full text Add to dashboard Cite

Polyphenols exhibit various beneficial biological activities and represent very promising candidates as active compounds for food industry. However, the low solubility, poor stability and low bioavailability of polyphenols have severely limited their industrial applications. enzymatic glycosylation is an effective way to improve the physicochemical properties of polyphenols. As efficient transglucosidases, glycoside hydrolase family 70 (GH70) glucansucrases naturally catalyze the synthesis of polysaccharides and oligosaccharides from sucrose. Notably, GH70 glucansucrases show broad acceptor substrate promiscuity and catalyze the glucosylation of a wide range of non-carbohydrate hydroxyl group-containing molecules, including benzenediol, phenolic acids, flavonoids and steviol glycosides. Branching sucrase enzymes, a newly established subfamily of GH70, are shown to possess a broader acceptor substrate binding pocket that acts efficiently for glucosylation of larger size polyphenols such as flavonoids. Here we present a comprehensive review of glucosylation of polyphenols using GH70 glucansucrase and branching sucrases. Their catalytic efficiency, the regioselectivity of glucosylation and the structure of generated products are described for these reactions. Moreover, enzyme engineering is effective for improving their catalytic efficiency and product specificity. The combined information provides novel insights on the glucosylation of polyphenols by GH70 glucansucrases and branching sucrases, and may promote their applications.

show abstract

“…The ratio of LCGS reached the highest level at pH 4, decreased to the lowest level at pH 5, and then gradually increased from pH 5 to 8 [ 50 ]. Son et al synthesized stevioside having low degree polymerized glucosides using dextransucrase and dextranase [ 51 ]. In this study, we improve the taste quality of glucosylated steviol glycoside through the protein engineering of CGTase.…”

Section: Resultsmentioning

confidence: 99%

Engineering of cyclodextrin glycosyltransferase improves the conversion efficiency of rebaudioside A to glucosylated steviol glycosides and increases the content of short-chain glycosylated steviol glycoside

et al. 2023

View full text Add to dashboard Cite

Background Compared with steviol glycosides, the taste of glucosylated steviol glycosides is better and more similar to that of sucrose. At present, cyclodextrin glucanotransferase (CGTase) is primarily used to catalyze the conversion of steviol glycosides to glucosylated steviol glycosides, with soluble starch serving as a glycosyl donor. The main disadvantages of enzymatic transglycosylation are the limited number of enzymes available, the low conversion rates that result in low yields, and the lack of selectivity in the degree of glycosylation of the products. In order to fill these gaps, the proteome of Alkalihalobacillus oshimensis (also named Bacillus oshimensis) was used for mining novel CGTases. Results Here, CGTase-15, a novel β-CGTase with a wide pH adaptation range, was identified and characterized. The catalyzed product of CGTase-15 tasted better than that of the commercial enzyme (Toruzyme® 3.0 L). In addition, two amino acid sites, Y199 and G265, which play important roles in the conversion of steviol glycosides to glucosylated steviol glycosides were identified by site-directed mutagenesis. Compared with CGTase-15, CGTase-15-Y199F mutant significantly increased the conversion rate of rebaudioside A (RA) to glucosylated steviol glycosides. Compared with CGTase-15, the content of short-chain glycosylated steviol glycosides catalyzed by CGTase-15-G265A mutant was significantly increased. Moreover, the function of Y199 and G265 was verified in other CGTases. The above mutation pattern has also been applied to CGTase-13 (a CGTase discovered by our laboratory with great potential in the production of glycosylated steviol glycosides), confirming that the catalytic product of CGTase-13-Y189F/G255A mutant has a better taste than that of CGTase-13. Conclusions This is the first report on the improvement of the sensory profiles of glycosylated steviol glycosides through site-directed mutagenesis of CGTase, which is significant for the production of glycosylated steviol glycosides.

show abstract

Synthesis and characterization of stevioside having low degree polymerized glucosides using dextransucrase and dextranase

Cited by 13 publications

References 44 publications

β-Glucosidase and β-Galactosidase-Mediated Transglycosylation of Steviol Glycosides Utilizing Industrial Byproducts

β-Glucosidase and β-Galactosidase-Mediated Transglycosylation of Steviol Glycosides Utilizing Industrial Byproducts

Enzymatic glucosylation of polyphenols using glucansucrases and branching sucrases of glycoside hydrolase family 70

Engineering of cyclodextrin glycosyltransferase improves the conversion efficiency of rebaudioside A to glucosylated steviol glycosides and increases the content of short-chain glycosylated steviol glycoside

Contact Info

Product

Resources

About