Purification and biochemical characterization of a mycelial glucose- and xylose-stimulated β-glucosidase from the thermophilic fungus Humicola insolens

Souza, Flávio Henrique Moreira; Nascimento, César Vanderlei; Rosa, José César; Masui, Douglas Chodi; Leone, Francisco de Assis; Jorge, João Atı́lio; Furriel, Rosa dos Prazeres Melo

doi:10.1016/j.procbio.2009.09.018

Cited by 77 publications

(58 citation statements)

References 40 publications

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“…by Kaur et al (2007). The K m and V max values for pNPG hydrolysis for Humicola insolens β-glucosidase were calculated to be 0.16 mM and 18.1 U/mg respectively (Souza et al, 2010).…”

Section: Enzyme Characterizationmentioning

confidence: 99%

Metabolic engineering and thermodynamic characterization of an extracellular -glucosidase produced by Aspergillus niger

Zahoor¹,

Javed²,

Aftab³

et al. 2011

Afr. J. Biotechnol.

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The production of an extracellular β-glucosidase by Aspergillus niger NRRL 599 was optimized using submerged fermentation technique. Effect of different media, different carbon sources, initial pH of the fermentation medium, temperature, incubation period and inoculum size on the production of β-glucosidase enzyme was investigated. A. niger NRRL 599 produced maximum extracellular β-glucosidase (4.48 U/mg) in Eggins and Pugh medium with 1% wheat bran (w/v) at pH 5.5 inoculated with 4% conidial suspension after 96 h of incubation at 30°C. Purified β-glucosidase gave K m and V max values of 3.11 mM and 20.83 U/mg respectively for pNPG hydrolysis. The enzyme was optimally active at pH 4.8 and at temperature of 60°C. Thermodynamic parameters, E a , ∆H and ∆S were found to be 52.17 KJ/mol, 49.90 J/mol.K and -71.69 KJ/mol, respectively. The pKa 1 and pKa 2 of ionizable groups of active site residues involved in V max were calculated to be 4.1 and 6.0 respectively.

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“…by Kaur et al (2007). The K m and V max values for pNPG hydrolysis for Humicola insolens β-glucosidase were calculated to be 0.16 mM and 18.1 U/mg respectively (Souza et al, 2010).…”

Section: Enzyme Characterizationmentioning

confidence: 99%

Metabolic engineering and thermodynamic characterization of an extracellular -glucosidase produced by Aspergillus niger

Zahoor¹,

Javed²,

Aftab³

et al. 2011

Afr. J. Biotechnol.

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“…Recent studies have shown that GH1 and GH3 β-glucosidases vary in their tolerance to glucose. In comparison to the GH3 β-glucosidases that have IC 50 values of less than 100 mM [6, 16, 36] due to the competitive inhibition by glucose [5, 12, 31], the GH1 counterparts have an IC 50 value of several hundreds to thousands of millimole [28, 33, 35]. A few studies have been carried out to reveal the high glucose tolerance mechanism of GH1 β-glucosidases.…”

Section: Introductionmentioning

confidence: 99%

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

Cao

Zhang

Shi

et al. 2018

Journal of Industrial Microbiology and Biotechnology

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In the feed industry, β-glucosidase has been widely used in the conversion of inactive and bounded soybean isoflavones into active aglycones. However, the conversion is frequently inhibited by the high concentration of intestinal glucose in monogastric animals. In this study, a GH1 β-glucosidase (AsBG1) with high specific activity, thermostability and glucose tolerance (IC50 = 800 mM) was identified. It showed great glucose tolerance against substrates with hydrophobic aryl ligands (such as pNPG and soy isoflavones). Using soybean meal as the substrate, AsBG1 exhibited higher hydrolysis efficiency than the GH3 counterpart Bgl3A with or without the presence of glucose in the reaction system. Furthermore, it is the first time to find that the endogenous β-glucosidase of soybean meal, mostly belonging to GH3, plays a role in the hydrolysis of soybean isoflavones and is highly sensitive to glucose. These findings lead to a conclusion that the GH1 rather than GH3 β-glucosidase has prosperous application advantages in the conversion of soybean isoflavones in the feed industry.Electronic supplementary materialThe online version of this article (10.1007/s10295-018-2040-6) contains supplementary material, which is available to authorized users.

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“…18 In order to readily screen conditions for optimal enzyme function, including temperature and pH, new methods are needed; ideally these methods would circumvent the need to express, purify, and identify optimal conditions on a per enzyme basis as done previously. 29–31 To identify an optimal pH for maximum broad glycoside hydrolase enzyme activity in the secretome from three T. reesei strains, QM6a, RUT-C30, and NG14, we utilized an in-gel fluorescent probe approach. Probe GH-ABP (Fig.…”

Section: Resultsmentioning

confidence: 99%

Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

et al. 2013

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Lignocellulosic biomass has great promise as a highly abundant and renewable source for the production of biofuels. However, the recalcitrant nature of lignocellulose toward hydrolysis into soluble sugars remains a significant challenge to harnessing the potential of this source of bioenergy. A primary method for deconstructing lignocellulose is via chemical treatments, high temperatures, and hydrolytic enzyme cocktails, many of which are derived from the fungus Trichoderma reesei. Herein, we use an activity-based probe for glycoside hydrolases to rapidly identify optimal conditions for maximum enzymatic lignocellulose deconstruction. We also demonstrate that subtle changes to enzyme composition and activity in various strains of T. reesei can be readily characterized by our probe approach. The approach also permits multimodal measurements, including fluorescent gel-based analysis of activity in response to varied conditions and treatments, and mass spectrometry-based quantitative identification of labelled proteins. We demonstrate the promise this probe approach holds to facilitate rapid production of enzyme cocktails for high-efficiency lignocellulose deconstruction to accommodate high-yield biofuel production.

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Purification and biochemical characterization of a mycelial glucose- and xylose-stimulated β-glucosidase from the thermophilic fungus Humicola insolens

Cited by 77 publications

References 40 publications

Metabolic engineering and thermodynamic characterization of an extracellular -glucosidase produced by Aspergillus niger

Metabolic engineering and thermodynamic characterization of an extracellular -glucosidase produced by Aspergillus niger

A highly glucose-tolerant GH1 β-glucosidase with greater conversion rate of soybean isoflavones in monogastric animals

Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

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