Synthesis of rutinosides and rutinose by reverse hydrolysis catalyzed by fungal α-<scp>l</scp>-rhamnosidases

Martearena, Maria Rita; Daz, Mirta Elizabeth; Ellenrieder, G.

doi:10.1080/10242420701568617

Cited by 17 publications

(13 citation statements)

References 18 publications

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“…Attack on oligosaccharides, such as fructans, and phenols may lead to the formation of phenol and sugar radicals that either form sugar–phenol compounds, repolymerized oligosaccharides or polymerized phenols [41]. Rutin might also arise in such a scenario—rutinose is not known to occur as free sugar in nature [42] and fungal enzymes have been suggested to be involved in its biosynthesis [43]. Only the 16 h variant of the deoxyribose degradations assay reveals that rutin, in comparison to quercetin, might be a more efficient antioxidant when it is present during the entire Haber–Weiss reaction.…”

Section: Resultsmentioning

confidence: 99%

Versatile Redox Chemistry Complicates Antioxidant Capacity Assessment: Flavonoids as Milieu-Dependent Anti- and Pro-Oxidants

Chobot

Kubicová

Bachmann

et al. 2013

IJMS

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Some antioxidants have been shown to possess additional pro-oxidant effects. Diverse methodologies exist for studying redox properties of synthetic and natural chemicals. The latter are substantial components of our diet. Exploration of their contribution to life-extending or -compromising effects is mandatory. Among reactive oxygen species (ROS), hydroxyl radical (•OH) is the most damaging species. Due to its short half-life, the assay has to contain a specific generation system. Plants synthesize flavonoids, phenolic compounds recognized as counter-agents to coronary heart disease. Their antioxidant activities are affected by their hydroxylation patterns. Moreover, in the plant, they mainly occur as glycosides. We chose three derivatives, quercetin, luteolin, and rutin, in attempts to explore their redox chemistry in contrasting hydrogen peroxide environments. Initial addition of hydrogen peroxide in high concentration or gradual development constituted a main factor affecting their redox chemical properties, especially in case of quercetin. Our study exemplifies that a combination of a chemical assay (deoxyribose degradation) with an electrochemical method (square-wave voltammetry) provides insightful data. The ambiguity of the tested flavonoids to act either as anti- or pro-oxidant may complicate categorization, but probably contributed to their evolution as components of a successful metabolic system that benefits both producer and consumer.

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Section: Resultsmentioning

confidence: 99%

Versatile Redox Chemistry Complicates Antioxidant Capacity Assessment: Flavonoids as Milieu-Dependent Anti- and Pro-Oxidants

Chobot

Kubicová

Bachmann

et al. 2013

IJMS

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“…It is a biotechnologically important enzyme in food industry and in the preparation of drugs and drug precursors (de Araujo et al 2013;Yadav et al 2010;You et al 2010). Meanwhile, some α-L-rhamnosidases can catalyze the synthesis of rhamnose-containing chemicals (RCCs) by reverse hydrolysis with the L-rhamnose as a donor (De Winter et al 2013;Lu et al 2015;Martearena et al 2008), suggesting that these enzymes would be attractive for synthesizing rare rhamnosylated compounds for chemical and pharmaceutical applications. Despite their industrial importance, few attempts have been made using protein engineering method to understand the catalytic mechanism for optimizing the catalytic efficiency of the existing rhamnosidases.…”

mentioning

confidence: 99%

“…Therefore, reverse hydrolysis has been considered as an economically feasible approach for industrial applications (Julio and Andrew 2007). Several glycosidases including α-rhamnosidase (Lu et al 2015;Martearena et al 2008), β-glucosidase (Vic et al 1997), β-galactosidase (Majumder et al 2008;Wei et al 2013), α-mannosidase (Maitin et al 2004;Singh et al 2000), and endo-α-N-acetylgalactosaminidase (Ashida et al 2010) have been used for glycoside synthesis through reverse hydrolysis reaction. It has been experimentally proved that the transglycosylation efficiency could be improved by mutagenesis of the glycosidases, and the sitedirected mutagenesis based on rational protein design is regarded as the common strategy to improve the catalytic properties in glycosidases on the basis of the solved homological 3D structures (Choi et al 2008;Hansson et al 2001;Hinz et al 2006;Lundemo et al 2013;Pollet et al 2010;Sun et al 2014;Zeuner et al 2014).…”

mentioning

confidence: 99%

Site-directed mutagenesis of α-l-rhamnosidase from Alternaria sp. L1 to enhance synthesis yield of reverse hydrolysis based on rational design

Liu

Yin³

et al. 2016

Appl Microbiol Biotechnol

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The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp, Asp, Asp, Glu, Arg, His, and Trp) and may form the hydrophobic pocket in stabilizing donor (Trp, Tyr, Tyr, and Trp) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp. Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme.

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“…It has been used in debittering of citrus, manufacturing of prunin from naringin, enhancing of wine aromas, and producing of L-rhamnose (Yadav, Yadav, Yadav, & Yadav, 2010). Moreover, it is a biotechnologically important enzyme in food industry in the preparation of drug and drug precursors, and some α-l-rhamnosidases can catalyze the synthesis of rhamnose-containing chemicals by reverse hydrolysis (RCCs) with the L-rhamnose as a donor (Chin, Murad, Mahadi, Nathan, & Bakar, 2013;Martearena, Daz, & Ellenrieder, 2009;Xu et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Enhancement of the thermostability of Aspergillus niger α‐ l ‐rhamnosidase based on PoPMuSiC algorithm

et al. 2019

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α‐l‐Rhamnosidase is a biotechnologically important enzyme in food industry and in the preparation of drugs and drug precursors. To expand the functionality of our previously cloned α‐l‐rhamnosidase from Aspergillus niger JMU‐TS528, 14 mutants were constructed based on the changes of the folding free energy (ΔΔG), predicted by the PoPMuSiC algorithm. Among them, six single‐site mutants displayed higher thermal stability than wild type (WT). The combinational mutant K573V–E631F displayed even higher thermostability than six single‐site mutants. The spectra analyses displayed that the WT and K573V–E631F had almost similar secondary and tertiary structure profiles. The simulated protein structure‐based interaction analysis and molecular dynamics calculation were further implemented to assess the conformational preferences of the K573V–E631F. The improved thermostability of mutant K573V–E631F may be attributed to the introduction of new cation–π and hydrophobic interactions, and the overall improvement of the enzyme conformation. Practical applications The stability of enzymes, particularly with regards to thermal stability remains a critical issue in industrial biotechnology and industrial processing generally tends to higher ambient temperature to inhibit microbial growth. Most of the α‐l‐rhamnosidases are usually active at temperature from 30 to 60°C, which are apt to denature at temperatures over 60°C. To expand the functionality of our previously cloned α‐l‐rhamnosidase from Aspergillus niger JMU‐TS528, we used protein engineering methods to increase the thermal stability of the α‐l‐rhamnosidase. Practically, conducting reactions at high temperatures enhances the solubility of substrates and products, increases the reaction rate thus reducing the reaction time, and inhibits the growth of contaminating microorganisms. Thus, the improvement on the thermostability of α‐l‐rhamnosidase on the one hand can increase enzyme efficacy; on the other hand, the high ambient temperature would enhance the solubility of natural substrates of α‐l‐rhamnosidase, such as naringin, rutin, and hesperidin, which are poorly dissolved in water at room temperature. Protein thermal resistance is an important issue beyond its obvious industrial importance. The current study also helps in the structure–function relationship study of α‐l‐rhamnosidase.

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Synthesis of rutinosides and rutinose by reverse hydrolysis catalyzed by fungal α-l-rhamnosidases

Cited by 17 publications

References 18 publications

Versatile Redox Chemistry Complicates Antioxidant Capacity Assessment: Flavonoids as Milieu-Dependent Anti- and Pro-Oxidants

Versatile Redox Chemistry Complicates Antioxidant Capacity Assessment: Flavonoids as Milieu-Dependent Anti- and Pro-Oxidants

Site-directed mutagenesis of α-l-rhamnosidase from Alternaria sp. L1 to enhance synthesis yield of reverse hydrolysis based on rational design

Enhancement of the thermostability of Aspergillus niger α‐ l ‐rhamnosidase based on PoPMuSiC algorithm

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