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. 2017 Dec;32(1):468-477.
doi: 10.1080/14756366.2016.1261131.

Purification and biochemical characterization of a novel thermostable serine alkaline protease from Aeribacillus pallidus C10: a potential additive for detergents

Vildan Yildirim  1 , Mustafa Ozkan Baltaci  1 , Ilknur Ozgencli  2 , Melda Sisecioglu  1 , Ahmet Adiguzel  1 , Gulsah Adiguzel  3
Affiliations

Purification and biochemical characterization of a novel thermostable serine alkaline protease from Aeribacillus pallidus C10: a potential additive for detergents

Vildan Yildirim et al. J Enzyme Inhib Med Chem. 2017 Dec.

Abstract

An extracellular thermostable alkaline serine protease enzyme from Aeribacillus pallidus C10 (GenBank No: KC333049), was purified 4.85 and 17. 32-fold with a yield of 26.9 and 19.56%, respectively, through DE52 anion exchange and Probond affinity chromatography. The molecular mass of the enzyme was determined through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), with approximately 38.35 kDa. The enzyme exhibited optimum activity at pH 9 and at temperature 60 °C. It was determined that the enzyme had remained stable at the range of pH 7.0-10.0, and that it had preserved more than 80% of its activity at a broad temperature range (20-80 °C). The enzyme activity was found to retain more than 70% and 55% in the presence of organic solvents and commercial detergents, respectively. In addition, it was observed that the enzyme activity had increased in the presence of 5% SDS. KM and Vmax values were calculated as 0.197 mg/mL and 7.29 μmol.mL-1.min-1, respectively.

Keywords: Aeribacillus pallidus; Thermotolerant; alkaline serine protease; biochemical characterization; purification.

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Figures

None
Graphical abstract
Figure 1.
Figure 1.
Neighbor-joining phylogenetic tree on the basis of 16S rRNA gene sequence data of the thermophilic bacteria. Alicyclobacillus acidiphilus DSM 14558 was used as out-group. Bootstrap values based on 1000 replications are listed as percentages at branching points. Only bootstrap values >50% are shown at nodes. The scale bar represented 1% divergence.
Figure 2.
Figure 2.
SDS-PAGE of the purified protease from A. pallidus C10. (A) Lane 1: Standard protein molecular mass markers, lane 2: (NH4)2SO4 precipitated proteins, lane 3–4: Purified protease from DE52 anion exchange chromatography, (B) Lane 1–2: Purified enzyme from Probond affinity chromatography, lane 3: Zymography of the purified enzyme.
Figure 3.
Figure 3.
Effect of pH on activity (A) and stability (B) of the purified protease from A. pallidus C10.
Figure 4.
Figure 4.
Effect of temperature on activity (A) and stability (B) of the purified protease from A. pallidus C10.
Figure 5.
Figure 5.
Stability of the alkaline protease from A. pallidus C10 in the presence of various commercial detergents.
See this image and copyright information in PMC

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