2013
DOI: 10.1093/abbs/gmt061
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Expression and characterization of a novel highly glucose-tolerant β-glucosidase from a soil metagenome

Abstract: A b-glucosidase gene unbgl1A was isolated by the functionbased screening of a metagenomic library and the enzyme protein was expressed in Escherichia coli, purified, and biochemically characterized. The enzyme Unbgl1A had a K m value of 2.09 + + + + + 0.31 mM, and a V max value of 183.90 + + + + + 9.61 mmol min 21 mg 21 under the optimal reaction conditions, which were pH 6.0 at 508 8 8 8 8C. Unbgl1A can be activated by a variety of monosaccharides, disaccharides, and NaCl, and exhibits a high level of stabili… Show more

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Cited by 50 publications
(20 citation statements)
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“…yunnanensis 258 ± 5.1182 ± 3.42.2 and 1.4 for glucose and xylose, respectively842 ± 15.560~10 h at 60 °C;BacteriumPresent work S. thermophilum 204 ± 12191 ± 140.7 and 0.5 for glucose and xylose, respectively8.96020 min at 55 °CFungi[8] H. insolens 180 ± 9200 ± 110.436.46044 min at 55 °CFungi[7] H. grisea var. thermoidea 1882020.529.5507 min at 60 °CFungi[23] B. halodurans 135.0 ± 4.3131.0 ± 0.2~0.8-0.9935030 min at 50 °CBacterium[24]Uncultured bacterium1201500.3 for glucose183.950 ND Soil metagenome[5] Marinomonas sp. MWYL1~132~1450.638940~11 min at 45 °CBacterium[9]

ND not determined

a One hundred percent specific activity (control) was estimated in the absence of carbohydrates

…”
Section: Discussionmentioning
confidence: 99%
“…yunnanensis 258 ± 5.1182 ± 3.42.2 and 1.4 for glucose and xylose, respectively842 ± 15.560~10 h at 60 °C;BacteriumPresent work S. thermophilum 204 ± 12191 ± 140.7 and 0.5 for glucose and xylose, respectively8.96020 min at 55 °CFungi[8] H. insolens 180 ± 9200 ± 110.436.46044 min at 55 °CFungi[7] H. grisea var. thermoidea 1882020.529.5507 min at 60 °CFungi[23] B. halodurans 135.0 ± 4.3131.0 ± 0.2~0.8-0.9935030 min at 50 °CBacterium[24]Uncultured bacterium1201500.3 for glucose183.950 ND Soil metagenome[5] Marinomonas sp. MWYL1~132~1450.638940~11 min at 45 °CBacterium[9]

ND not determined

a One hundred percent specific activity (control) was estimated in the absence of carbohydrates

…”
Section: Discussionmentioning
confidence: 99%
“…For examples, β-glucosidase from Microbispora bispora was activated in the presence of 300 mM glucose [135] and that reported in T. thermosaccharolyticum DSM 571 and T. aotearoense was activated by glucose concentration < 200 and 250 mM, respectively [85,136]. Similarly, recombinant GH 1 β-glucosidase from Exiguobacterium antarcticum B7 [84], Fervidobacterium islandicum [137], Thermotoga thermarum DSM 5069T [138] and Anoxybacillus DT3-1 [83] was tolerant to high glucose concentration e.g., Ki of 0.54-15 M. On the other hand, glucose tolerant β-glucosidase with Ki of 1.0-4.8 M has been identified from different environments using metagenomic approaches [45,53,106,111,139,140].…”
Section: Glucose Sensitivity and Tolerancementioning
confidence: 99%
“…Cocktails have generally been based on commercial enzymes isolated from different microorganisms, which may have different optimal growth conditions. Recently, the isolation of novel enzymes, including thermostable enzymes from metagenomic libraries, has been extensively studied, leading to the isolation of many enzymes with special characteristics such as being cold- (Wierzbicka-Wos et al, 2013 ) or heat-active (Schroder et al, 2014 ), and ionic liquid- (Ilmberger et al, 2012 ) or product-tolerant (Lu et al, 2013 ). Microorganisms in the same metagenome generally live together for a relatively long time, and should have similar tolerance for environmental conditions, such as temperature, pH, and oxidation-reduction potentials, which may result in isolation of enzymes with similar optimal conditions.…”
Section: Discussionmentioning
confidence: 99%