Identification of an active-site residue in yeast invertase by affinity labeling and site-directed mutagenesis.

Reddy, V.Anthony; Maley, Frank

doi:10.1016/s0021-9258(19)38518-7

Cited by 90 publications

(10 citation statements)

References 37 publications

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“…Despite co-option from distinct lineages, the two independently evolved ASFFs are both trichome-enriched, neofunctionalized GH32 β-fructofuranosidases, which lack the canonical amino acid sequence DDTK sucrose-binding pocket of Arabidopsis cell-wall invertase 1 (AT3G13790; fig. S4) ( 40 – 42 ). This is consistent with the observation that neither ASFF uses unacylated sucrose as substrate (fig.…”

Section: Discussionmentioning

confidence: 99%

It happened again: Convergent evolution of acylglucose specialized metabolism in black nightshade and wild tomato

et al. 2021

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Section: Discussionmentioning

confidence: 99%

It happened again: Convergent evolution of acylglucose specialized metabolism in black nightshade and wild tomato

et al. 2021

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“…This is a perfect match for the prosite pattern HX # PX % [L\I\V\M]NDPNG, which is indicative of the membership of the two enzymes in glycoside hydrolase family 32 [27]. All residues that resemble those in related enzymes (on the basis of structural predictions [28]) that have earlier been shown to be involved directly in the catalytic process [29][30][31][32] are conserved in both Aspergillus enzymes. These are, in particular, Asp-41, Asp-189 and Glu-241.…”

Section: All Datamentioning

confidence: 52%

Purification, characterization, gene cloning and preliminary X-ray data of the exo-inulinase from Aspergillus awamori

et al. 2002

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Extracellular exo-inulinase has been isolated from a solid-phase culture of the filamentous fungus Aspergillus awamori var. 2250. The apparent molecular mass of the monomer enzyme was 69 +/- kDa, with a pI of 4.4 and a pH optimum of 4.5. The enzyme hydrolysed the beta-(2-->1)-fructan (inulin) and beta-(2-->6)-fructan (levan) via exo-cleavage, releasing fructose. The values for the Michaelis constants K(m) and V(max) in the hydrolysis of inulin were 0.003 +/- 0.0001 mM and 175 +/- 5 micromol.min(-1).mg(-1). The same parameters in the hydrolysis of levan were 2.08 +/- 0.04 mg/ml and 1.2 +/- 0.02 micromol/min per mg, respectively. The gene and cDNA encoding the A. awamori exo-inulinase were cloned and sequenced. The amino acid sequence indicated that the protein belongs to glycoside hydrolase family 32. A surprisingly high similarity was found to fructosyltransferase from Aspergillus foetidus (90.7% on the level of the amino acid sequence), despite the fact that the latter enzyme is unable to hydrolyse inulin and levan. Crystals of the native exo-inulinase were obtained and found to belong to the orthorhombic space group P2(1)2(1)2(1) with cell parameters a=64.726 A (1A=0.1 nm), b=82.041 A and c=136.075 A. Crystals diffracted beyond 1.54 A, and useful X-ray data were collected to a resolution of 1.73 A.

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“…From the comparison between members of the GH32 family, the catalytic dyad aspartate (Asp47) stands out as a nucleophile, 65 whereas the glutamate (Glu222) acts as the general acid/base catalyst, 66 typical for GH32. Tyrosine (Tyr287) was suggested by Verhaest et al (2005) 52 to be involved in the pK a modulation of the equivalent to Glu222 acid/base catalyst because of its proximity, while Yuan et al (2012) 67 showed that the arginine equivalent to Arg166 could be a more suitable candidate based on theoretical pK a calculations and molecular dynamics simulations.…”

Section: Resultsmentioning

confidence: 99%

Crystal Structure of a Sucrose-6-phosphate Hydrolase from Lactobacillus gasseri with Potential Applications in Fructan Production and the Food Industry

Lima

Almeida

Mera

et al. 2021

J. Agric. Food Chem.

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Fructooligosaccharides (FOSs) are polymers of fructose with a prebiotic activity because of their production and fermentation by bacteria that inhabit the gastrointestinal tract and are widely used in the industry and new functional foods. Lactobacillus gasseri stands out as an important homofermentative microorganism related to FOS production, and its potential applications in the industry are undeniable. In this study, we report the production and characterization of a sucrose-6-phosphate hydrolase from L. gasseri belonging to the GH32 family. Apo-LgAs32 and LgAs32 complexed with β-d-fructose structures were determined at a resolution of 1.94 and 1.84 Å, respectively. The production of FOS, fructans, 1-kestose, and nystose by the recombinant LgAs32, using sucrose as a substrate, shown in this study is very promising. When compared to its homologous enzyme from Lactobacillus reuteri, the production of 1-kestose by LgAs32 is increased; thus, LgAs32 can be considered as an alternative in fructan production and other industrial applications.

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Identification of an active-site residue in yeast invertase by affinity labeling and site-directed mutagenesis.

Cited by 90 publications

References 37 publications

It happened again: Convergent evolution of acylglucose specialized metabolism in black nightshade and wild tomato

It happened again: Convergent evolution of acylglucose specialized metabolism in black nightshade and wild tomato

Purification, characterization, gene cloning and preliminary X-ray data of the exo-inulinase from Aspergillus awamori

Crystal Structure of a Sucrose-6-phosphate Hydrolase from Lactobacillus gasseri with Potential Applications in Fructan Production and the Food Industry

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