Molecular characterization and heterologous expression of a Xanthophyllomyces dendrorhous α-glucosidase with potential for prebiotics production

Gutiérrez-Alonso, Patricia; Gimeno-Pérez, María; Ramirez-Escudero, M.; Plou, Francisco J.; Sanz‐Aparicio, Julia; Fernández-Lobato, María

doi:10.1007/s00253-015-7171-3

Cited by 22 publications

(17 citation statements)

References 35 publications

(49 reference statements)

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“…By 24 h of reaction with 500 mM maltose, the amount of transglycosylation products was 12.6% of total sugars in the reaction mixture, while the respective value for the MAL62 protein was about three times less-4.5% (see Table S3). Many α-glucosidases can transglycosylate and produce short oligosaccharides, especially at high concentration of the substrate [17][18][19][20][21]. We assayed this possibility by incubating BaAG2 with 250 and 500 mM maltose up to 72 h and analyzed the reaction products by HPLC.…”

Section: The Transglycosylation Of Maltosementioning

confidence: 99%

“…Phaffia rhodozyma) has been studied in detail, and synthesis of tri-and tetrasaccharides with α-1,6 linkages was detected. This enzyme certainly has a biotechnological potential-it produced 3.6 times more transglycosylation products than the S. cerevisiae α-glucosidase studied at the same conditions [17,20].…”

Section: Introductionmentioning

confidence: 99%

“…α-Glucosidases have been popular objects to study protein evolution and phylogenesis [13][14][15][16], but they also have a biotechnological value. Thus, some of them have a high transglycosylating activity thanks to which they produce prebiotic oligosaccharides and potential functional food ingredients such as panose, melezitose, isomelezitose and isomalto-oligosaccharides [17][18][19][20][21]. For example, the α-glucosidase of S. cerevisiae produced isomelezitose from sucrose when the substrate concentration was high [21].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of a Maltase from an Early-Diverged Non-Conventional Yeast Blastobotrys adeninivorans

Visnapuu

Meldre

Põšnograjeva

et al. 2019

IJMS

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Genome of an early-diverged yeast Blastobotrys (Arxula) adeninivorans (Ba) encodes 88 glycoside hydrolases (GHs) including two α-glucosidases of GH13 family. One of those, the rna_ARAD1D20130g-encoded protein (BaAG2; 581 aa) was overexpressed in Escherichia coli, purified and characterized. We showed that maltose, other maltose-like substrates (maltulose, turanose, maltotriose, melezitose, malto-oligosaccharides of DP 4‒7) and sucrose were hydrolyzed by BaAG2, whereas isomaltose and isomaltose-like substrates (palatinose, α-methylglucoside) were not, confirming that BaAG2 is a maltase. BaAG2 was competitively inhibited by a diabetes drug acarbose (Ki = 0.8 µM) and Tris (Ki = 70.5 µM). BaAG2 was competitively inhibited also by isomaltose-like sugars and a hydrolysis product—glucose. At high maltose concentrations, BaAG2 exhibited transglycosylating ability producing potentially prebiotic di- and trisaccharides. Atypically for yeast maltases, a low but clearly recordable exo-hydrolytic activity on amylose, amylopectin and glycogen was detected. Saccharomyces cerevisiae maltase MAL62, studied for comparison, had only minimal ability to hydrolyze these polymers, and its transglycosylating activity was about three times lower compared to BaAG2. Sequence identity of BaAG2 with other maltases was only moderate being the highest (51%) with the maltase MalT of Aspergillus oryzae.

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Section: The Transglycosylation Of Maltosementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of a Maltase from an Early-Diverged Non-Conventional Yeast Blastobotrys adeninivorans

Visnapuu

Meldre

Põšnograjeva

et al. 2019

IJMS

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“…It was extensively reported that the hydrolytic activity of glycosidases, such as α/β‐glucosidases or β‐fructofuranosidases, could be shifted towards synthesis (trasnsglycosylating activity) using a high substrate concentration. Thus, the α‐glucosidase from the yeast Xanthophyllomyces dendrorhous produced IMOS (mainly panose) using 200–525 g l −1 maltose (Fernández‐Arrojo et al , ; Gutierrez‐Alonso et al , ) and β‐fructofuranosidases from Aspergillus awamori or yeast such as Schwanniomyces occidentalis and Saccharomyces cerevisiae synthesized FOS using different sucrose‐rich by‐products from a variety of palm tree dates (the first) or 600 g l −1 sucrose (the last two) (Álvaro‐Benito et al , ; Lafraya et al , ; Smaali et al , ). In fact, sucrose is a relatively cheap and renewable raw material that has already been widely employed in the synthesis of bioactive oligosaccharides (Monsan and Ouarné, ), and among them several hetero‐oligosaccharides.…”

Section: Introductionmentioning

confidence: 99%

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

García-González

Plou

Cervantes

et al. 2019

Microbial Biotechnology

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Summary Metschnikowia reukaufii is a widespread yeast able to grow in the plants’ floral nectaries, an environment of extreme conditions with sucrose concentrations exceeding 400 g l−1, which led us into the search for enzymatic activities involved in this sugar use/transformation. New oligosaccharides were produced by transglucosylation processes employing M. reukaufii cell extracts in overload‐sucrose reactions. These products were purified and structurally characterized by MS‐ESI and NMR techniques. The reaction mixture included new sugars showing a great variety of glycosidic bonds including α‐(1→1), α‐(1→3) and α‐(1→6) linkages. The main product synthesized was the trisaccharide isomelezitose, whose maximum concentration reached 81 g l−1, the highest amount reported for any unmodified enzyme or microbial extract. In addition, 51 g l−1 of the disaccharide trehalulose was also produced. Both sugars show potential nutraceutical and prebiotic properties. Interestingly, the sugar mixture obtained in the biosynthetic reactions also contained oligosaccharides such as esculose, a rare trisaccharide with no previous NMR structure elucidation, as well as erlose, melezitose and theanderose. All the sugars produced are naturally found in honey. These compounds are of biotechnological interest due to their potential food, cosmeceutical and pharmaceutical applications.

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“…Further, the promoter for crtS contains four consensus motifs for the binding of CreA [13], which is a negative regulator involved in catabolic repression in Aspergillus nidulans [18]. In addition, extracellular α-glucosidase and invertase activities were not detected in X. dendrorhous cultures when glucose was used as a carbon source, suggesting the catabolic repression of these enzymes [19, 20]. This indicates that this regulatory mechanism is operative in X. dendrorhous , as genes encoding glycosyl hydrolases are well known as targets of catabolic repression.…”

Section: Introductionmentioning

confidence: 99%

Regulation of carotenogenesis in the red yeast Xanthophyllomyces dendrorhous: the role of the transcriptional co-repressor complex Cyc8–Tup1 involved in catabolic repression

et al. 2016

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BackgroundThe yeast Xanthophyllomyces dendrorhous produces carotenoids of commercial interest, including astaxanthin and β-carotene. Although carotenogenesis in this yeast and the expression profiles of the genes controlling this pathway are known, the mechanisms regulating this process remain poorly understood. Several studies have demonstrated that glucose represses carotenogenesis in X. dendrorhous, suggesting that this pathway could be regulated by catabolic repression. Catabolic repression is a highly conserved regulatory mechanism in eukaryotes and has been widely studied in Saccharomyces cerevisiae. Glucose-dependent repression is mainly observed at the transcriptional level and depends on the DNA-binding regulator Mig1, which recruits the co-repressor complex Cyc8–Tup1, which then represses the expression of target genes. In this work, we studied the regulation of carotenogenesis by catabolic repression in X. dendrorhous, focusing on the role of the co-repressor complex Cyc8–Tup1.ResultsThe X. dendrorhous CYC8 and TUP1 genes were identified, and their functions were demonstrated by heterologous complementation in S. cerevisiae. In addition, cyc8 − and tup1 − mutant strains of X. dendrorhous were obtained, and both mutations were shown to prevent the glucose-dependent repression of carotenogenesis in X. dendrorhous, increasing the carotenoid production in both mutant strains. Furthermore, the effects of glucose on the transcript levels of genes involved in carotenogenesis differed between the mutant strains and wild-type X. dendrorhous, particularly for genes involved in the synthesis of carotenoid precursors, such as HMGR, idi and FPS. Additionally, transcriptomic analyses showed that cyc8 − and tup1 − mutations affected the expression of over 250 genes in X. dendrorhous. ConclusionsThe CYC8 and TUP1 genes are functional in X. dendrorhous, and their gene products are involved in catabolic repression and carotenogenesis regulation. This study presents the first report involving the participation of Cyc8 and Tup1 in carotenogenesis regulation in yeast.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0597-1) contains supplementary material, which is available to authorized users.

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Molecular characterization and heterologous expression of a Xanthophyllomyces dendrorhous α-glucosidase with potential for prebiotics production

Cited by 22 publications

References 35 publications

Characterization of a Maltase from an Early-Diverged Non-Conventional Yeast Blastobotrys adeninivorans

Characterization of a Maltase from an Early-Diverged Non-Conventional Yeast Blastobotrys adeninivorans

Efficient production of isomelezitose by a glucosyltransferase activity in Metschnikowia reukaufii cell extracts

Regulation of carotenogenesis in the red yeast Xanthophyllomyces dendrorhous: the role of the transcriptional co-repressor complex Cyc8–Tup1 involved in catabolic repression

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