Enzymatic Production of Steviol Glucosides Using β-Glucosidase and Their Applications

Nguyen, Thi Thanh Hanh; Seo, Changseop; Kwak, So Hyung; Kim, Jeesoo; Kang, Hee Kyoung; Kim, Seong Bo; Kim, Doman

doi:10.1016/b978-0-12-813280-7.00023-2

Cited by 13 publications

(9 citation statements)

References 73 publications

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“…Stevia stems and leaves are rich in diterpenoid glycosides, mainly stevioside and rebaudioside A (rebA), but many more minor glycosides have been isolated. The chemical structure of stevia glycosides includes the diterpenoid steviol, glycosylated through an ester bond at the C13 or C19 with one or more glucose moieties, connected with β-1,2 or β-1,3 glycosidic bond ( Nguyen et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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

Zerva

Chorozian

Kritikou

et al. 2021

Front. Bioeng. Biotechnol.

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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.

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Section: Introductionmentioning

confidence: 99%

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

Zerva

Chorozian

Kritikou

et al. 2021

Front. Bioeng. Biotechnol.

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“…Prírodné sladidlá sa v poslednej dobe tešia veľkému záujmu. Hlavným dôvodom je zvyšovanie zdravotných obáv z konzumácie cukru, ale aj problémy týkajúce sa bezpečnosti niektorých nevýživných umelých sladidiel (Nguyen et al, 2019).…”

Section: Discussionunclassified

Food safety and quality. Scientific proceedings

2023

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We tested 68 strains of Staphylococci, (S. aureus -29, S. epidermidis -34, S. haemolyticus -23, S. warneri -16, and S. chromogenes -6). They have been isolated from various types of mastitis. Staphylococci in milk samples were isolated from acute form (27,9 %), subacute form (10,3 %), subclinical form (25 %) and latent form (36,8 %). The most common form of mastitis in dairy cows was latent mastitis. Formation of biofilm was evaluated by results of two methods: quantitative growth on CongoRed agar (CRA) and semiquantitative growth on microtiter plates (MTP). At the significance level of α = 0.05, the dependence of biofilm formation in bacteria positive using both phenotypic methods in S. aureus, S. epidermidis and S. haemolyticus was confirmed. The positivity in S. warneri and S. chromogenes determined by both methods was random. Based on our results and found dependence in comparison both methods of biofilm detection, we can recommend the simultaneous use of these methods in ideal conditions. It should be supplemented by determining the presence of genes encoding biofilm production. Keywords production of biofil:, staphylococci m, cow´s mastitis, milk ÚVOD Tvorba biofilmu ako významný faktor virulencie pomáha baktériám vyhnúť sa imunitnej obrane hostiteľa a antimikrobiálnej terapii. V chovoch prežúvavcov má biofilm vplyv na intenzitu a pretrvávanie mastitíd spôsobených v prevažnej miere baktériami z rodu Staphylococcus spp., ktoré sú najčastejšou príčinou mastitíd u prežúvavcov (Vasiľ et al., 2017).Vzhľadom na úlohu biofilmu pri perzistencii a kolonizácii stafylokokov v epiteli mliečnej žľazy, zníženie imunologickej odpovede organizmu, rezistencie na antibiotickú liečbu a ťažkosti pri eradikácii patogénov má podrobné preskúmanie a pochopenie mechanizmu tvorby biofilmu zásadný význam pri terapii príp. prevencii zápalov mliečnej žľazy. Je všeobecne známe, že baktérie produkujúce biofilm je ťažké z mliečnej žľazy odstrániť, sú často rezistentné na antibiotickú liečbu, ktorá sa na liečbu mastitíd bežne používa (Farkašová et al., 2020).Cieľom našej práce bolo stanovenie biofilmu u stafylokokov izolovaných z mlieka mastitídnych dojníc počas jedného roka. MATERIÁL A METODIKAVyšetrované vzorky mlieka sme odobrali v priebehu jedného roka od dojníc na PD Kluknava na východe Slovenska. Odbery sme uskutočnili u dojníc v štvrťročných intervaloch (marec, jún, september december). Celkom bolo vyšetrených 120 kusov dobytka. Mlieko bolo hneď po odbere vyšetrené maštaľným NK testom na detekciu typu mastitídy (akútna, subakútna, subklinická, latentná), a následne bakteriologickou kultiváciou na 5 % krvnom agare a na špeciálnych selektívnych pôdach. Následne boli stafylokokové kmene identifikované pomocou dvoch rôznych identifikačných metód: na základe biochemických enzymatických vlastností baktérií STAPHYtest 24 s identifikačným programom TNW 7.0 (Erba-Lachema, Brno, Česká republika) s presnosťou detekcie viac ako 90,0 % a na základe stanovenia

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“…Previous studies on the hydrolysis and modification of steviol glycosides mainly focused on ST and RA, while there were few reports on RC. It has been reported that some β-glucosidase and β-galactosidase from microorganisms hydrolyze ST and RA to STE ( Nguyen et al, 2019 ), demonstrating their ability to hydrolyze the β-glucosidic linkage of the C13 side chain and β-glucosyl ester linkage of C19 side chain ( Chen et al, 2014 ; Nguyen et al, 2016 ), although the substrate specificity of enzymes from different sources varies greatly ( Lan et al, 2019 ; Yan et al, 2021 ). Based on sequence homology, the proteins encoded by 1932, 1940, 1941, and 1944 genes belong to β-glucosidase of glycoside hydrolase family 3.…”

Section: Discussionmentioning

confidence: 99%

Integrated metabolite profiling and transcriptome analysis unraveling mechanism of RC catabolism in Paenarthrobacter ilicis CR5301

Li¹,

Sun²,

Cao³

et al. 2023

Front. Microbiol.

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Steviol glycosides are ideal sweeteners that are widely used in food, medicine, and cosmetics. Rebaudioside C (RC) is considered to be the third most abundant steviol glycoside, which has a bitter aftertaste that limits its application. Hydrolysis of RC to generate other bioactive steviol glycosides is an effective way to promote its additional utilization. In our previous study, a bacterium Paenarthrobacter ilicis CR5301 was isolated and identified for hydrolyzing RC with high efficiency. Herein, the expression profiles of P. ilicis CR5301 in the deletion and presence of RC were investigated by RNA-seq. The RC metabolites were identified by high-performance liquid chromatography and ultra-performance liquid chromatography-triple-time of flight mass spectrometry. Novel results were discovered in four aspects of research. First, the identification of metabolites revealed that four metabolites, namely, dulcoside A, dulcoside B, dulcoside A1, and steviol, were produced during RC metabolism. Second, RNA-seq analyses unraveled that 105 genes of P. ilicis CR5301 were significantly differentially expressed, and 7 pathways were significantly enriched. Third, independent RT-qPCR verified the accuracy and reliability of the RNA-seq results. Finally, a complete catabolic model of RC in P. ilicis CR5301 was proposed, and key genes were indicated in the RC catabolic metabolism by combining them with literature and sequence alignments. This study comprehensively unraveled the genes and pathways of RC catabolism in P. ilicis CR5301 at the transcriptional and metabolic levels. It provided new insights and evidence for understanding the mechanism of RC catabolism in bacteria. Key candidate genes may potentially contribute to the RC hydrolysis and preparation of other functional steviol glycosides in the future.

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Enzymatic Production of Steviol Glucosides Using β-Glucosidase and Their Applications

Cited by 13 publications

References 73 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

Food safety and quality. Scientific proceedings

Integrated metabolite profiling and transcriptome analysis unraveling mechanism of RC catabolism in Paenarthrobacter ilicis CR5301

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