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. 2015 Oct 11:2015:978632.
doi: 10.1155/2015/978632. eCollection 2015.

A Novel Highly Thermostable Multifunctional Beta-Glycosidase from Crenarchaeon Acidilobus saccharovorans

Vadim M Gumerov  1 , Andrey L Rakitin  1 , Andrey V Mardanov  1 , Nikolai V Ravin  1
Affiliations

A Novel Highly Thermostable Multifunctional Beta-Glycosidase from Crenarchaeon Acidilobus saccharovorans

Vadim M Gumerov et al. Archaea. .

Abstract

We expressed a putative β-galactosidase Asac_1390 from hyperthermophilic crenarchaeon Acidilobus saccharovorans in Escherichia coli and 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 with p-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 multifunctional β-glycosidase. Therefore, the utilization of Asac_1390 may contribute to facilitating the efficient degradation of lignocellulosic biomass and help enhance bioconversion processes.

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Figures

Figure 1
Figure 1
Expression and purification of recombinant glycosidase Asac_1390. SDS-PAGE was done using a 10.0% polyacrylamide gel; proteins were stained with Coomassie Brilliant Blue R-250. Lanes: 1—molecular weight markers (sizes are shown in kDa); 2—total protein from uninduced cells; 3—total protein from induced cells; 4—purified recombinant Asac_1390.
Figure 2
Figure 2
Effects of pH (a) and temperature (b) on the β-galactosidase activity of recombinant Asac_1390. Data represent the means of three experiments and error bars represent standard deviation.
Figure 3
Figure 3
Thermal inactivation of recombinant Asac_1390. Purified Asac_1390 was preincubated at 90°C followed by β-galactosidase assay. Data represent the means of three experiments and error bars represent standard deviation.
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