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

2019 ◽  
Vol 21 (1) ◽  
pp. 297 ◽  
Author(s):  
Triinu Visnapuu ◽  
Aivar Meldre ◽  
Kristina Põšnograjeva ◽  
Katrin Viigand ◽  
Karin Ernits ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Aspergillus Oryzae ◽  
Hydrolysis Product ◽  
Hydrolytic Activity ◽  
Glycoside Hydrolases ◽  
Arxula Adeninivorans ◽  
Sequence Identity ◽  
Diabetes Drug

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.

Expression and characterization of an α-glucosidase from Thermoanaerobacter ethanolicus JW200 with potential for industrial application

Biologia ◽  
2009 ◽  
Vol 64 (6) ◽  
Author(s):  
Yue-Hong Wang ◽  
Yu Jiang ◽  
Zuo-Ying Duan ◽  
Wei-Lan Shao ◽  
Hua-Zhong Li
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Optimal Condition ◽  
Industrial Application ◽  
Conversion Rate ◽  
Hydrolytic Activity ◽  
Thermoanaerobacter Ethanolicus ◽  
Transglycosylation Activity

AbstractIn this study, a new α-glucosidase gene from Thermoanaerobacter ethanolicus JW200 was cloned and expressed in Escherichia coli by a novel heat-shock vector pHsh. The recombinant α-glucosidase exhibited its maximum hydrolytic activity at 70°C and pH 5.0∼5.5. With p-nitrophenyl-α-D-glucoside as a substrate and under the optimal condition (70°C, pH 5.5), K m and V max of the enzyme was 1.72 mM and 39 U/mg, respectively. The purified α-glucosidase could hydrolyze oligosaccharides with both α-1,4 and α-1,6 linkages. The enzyme also had strong transglycosylation activity when maltose was used as sugar donor. The transglucosylation products towards maltose are isomaltose, maltotriose, panose, isomaltotriose and tetrasaccharides. The enzyme could convert 400 g/L maltose to oligosaccharides with a conversion rate of 52%, and 83% of the oligosaccharides formed were prebiotic isomaltooligosaccharides (containing isomaltose, panose and isomaltotriose).

scholarly journals Cloning and expression of Saccharomyces cerevisiae copper-metallothionein gene in Escherichia coli and characterization of the recombinant protein

1993 ◽  
Vol 212 (2) ◽  
pp. 521-528 ◽  
Author(s):  
Zehra SAYERS ◽  
Patricia BROUILLON ◽  
Constantin E. VORGIAS ◽  
Hans F. NOLTING ◽  
Christoph HERMES ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Recombinant Protein ◽  
Cloning And Expression ◽  
Metallothionein Gene

Characterization of ubiquitin C-terminal hydrolase 1 (YUH1) from Saccharomyces cerevisiae expressed in recombinant Escherichia coli

2007 ◽  
Vol 56 (1) ◽  
pp. 20-26 ◽  
Author(s):  
Hyun-Ah Yu ◽  
Sung-Gun Kim ◽  
Eun-Jeong Kim ◽  
Woo-Jong Lee ◽  
Dae-Ok Kim ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Recombinant Escherichia Coli

Expression, purification, and characterization of asparaginase II from Saccharomyces cerevisiae in Escherichia coli

2019 ◽  
Vol 159 ◽  
pp. 21-26 ◽  
Author(s):  
Wagner Lopes ◽  
Bárbara Adriana Ferreira dos Santos ◽  
André Luiz Franco Sampaio ◽  
Ana Paula Gregório Alves Fontão ◽  
Hilton Jorge Nascimento ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Purification And Characterization

scholarly journals Characterization of TbPDE2A, a Novel Cyclic Nucleotide-specific Phosphodiesterase from the Protozoan ParasiteTrypanosoma brucei

2000 ◽  
Vol 276 (15) ◽  
pp. 11559-11566 ◽  
Author(s):  
Roya Zoraghi ◽  
Stefan Kunz ◽  
Kewei Gong ◽  
Thomas Seebeck
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Trypanosoma Brucei ◽  
Dictyostelium Discoideum ◽  
Cyclic Nucleotide ◽  
Catalytic Domain ◽  
Sequence Identity ◽  
Essential Enzyme ◽  
Identification And Characterization

This study reports the identification and characterization of a cAMP-specific phosphodiesterase from the parasitic hemoflagellateTrypanosoma brucei. TbPDE2A is a class I phosphodiesterase. Its catalytic domain exhibits 30–40% sequence identity with those of all 11 mammalian phosphodiesterase (PDE) families, as well as withPDE2fromSaccharomyces cerevisiae,duncefromDrosophila melanogaster, andregAfromDictyostelium discoideum. The overall structure of TbPDE2A resembles that of human PDE11A in that its N-terminal region contains a single GAF domain. This domain is very similar to those of the mammalian PDE2, -5, -6, -10, and -11, where it constitutes a potential cGMP binding site. TbPDE2A can be expressed inS. cerevisiae, and it complements anS. cerevisiaePDE deletion strain. Recombinant TbPDE2A is specific for cAMP, with aKmof ∼2 μm. It is entirely resistant to the nonselective PDE inhibitor 3-isobutyl-1-methylxanthine, but it is sensitive to trequinsin, dipyridamole, sildenafil, and ethaverine with IC50values of 5.4, 5.9, 9.4, and 14.2 μm, respectively. All four compounds inhibit proliferation of bloodstream form trypanosomes in culture, indicating that TbPDE2A is an essential enzyme.

PAPS-Reductase from Escherichia coli: Characterization of the Enzyme as Probe for Thioredoxins

1987 ◽  
Vol 42 (1-2) ◽  
pp. 93-102 ◽  
Author(s):  
Jens D . Schwenn ◽  
Ulrich Schriek
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Malate Dehydrogenase ◽  
Cross Reactivity ◽  
Enzymatic Assay ◽  
Fructose Bisphosphatase ◽  
Simple Method ◽  
Bacterial Enzyme ◽  
Eukaryotic Organisms

Abstract PAPS-reductase from Escherichia coli was em ployed to detect thioredoxins from pro- and eukaryotic organisms. A simple method for the isolation of this enzyme and properties of the enzymatic assay were described. A comparison betw een thioredoxins detected by the PAPS-reductase and the Fructose-bisphosphatase or NADP malate dehydrogenase was used to assess the validity of the test. The high cross-reactivity of the bacterial enzyme was useful in the purification of heterologous thioredoxins from spinach, Synechococcus, and Saccharomyces cerevisiae.

scholarly journals A Novel Highly Thermostable Multifunctional Beta-Glycosidase from CrenarchaeonAcidilobus saccharovorans

Archaea ◽  
2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Vadim M. Gumerov ◽  
Andrey L. Rakitin ◽  
Andrey V. Mardanov ◽  
Nikolai V. Ravin
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Half Life ◽  
Sequence Similarity ◽  
Hydrolytic Activity ◽  
Maximum Activity ◽  
Glycoside Hydrolases ◽  
Amino Acid Residues ◽  
High Tolerance ◽  
High Sequence Similarity

We expressed a putativeβ-galactosidase Asac_1390 from hyperthermophilic crenarchaeonAcidilobus saccharovoransinEscherichia coliand purified the recombinant enzyme. Asac_1390 is composed of 490 amino acid residues and showed high sequence similarity to family 1 glycoside hydrolases from various thermophilic Crenarchaeota. The maximum activity was observed at pH 6.0 and 93°C. The half-life of the enzyme at 90°C was about 7 hours. Asac_1390 displayed high tolerance to glucose and exhibits hydrolytic activity towards cellobiose and various aryl glucosides. The hydrolytic activity withp-nitrophenyl (pNP) substrates followed the order pNP-β-D-galactopyranoside (328 U mg−1), pNP-β-D-glucopyranoside (246 U mg−1), pNP-β-D-xylopyranoside (72 U mg−1), and pNP-β-D-mannopyranoside (28 U mg−1). Thus the enzyme was actually a multifunctionalβ-glycosidase. Therefore, the utilization of Asac_1390 may contribute to facilitating the efficient degradation of lignocellulosic biomass and help enhance bioconversion processes.

Sign in / Sign up

Export Citation Format

Share Document