title: Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification.
abstract: Abstract In this study, protease-producing capacity of Bacillus pumilus D3, isolated from hydrocarbon contaminated soil, was evaluated and optimized. Optimum growing conditions for B. pumilus D3 in terms of protease production were determined as 1% optimum inoculum size, 35 °C temperature, 11 pH and 48 h incubation time, respectively. Stability studies indicated that the mentioned protease was stable within the pH range of 7-10.5 and between 30 °C and 40 °C temperatures. Surprisingly, the activity of the enzyme increased in the presence of SDS with concentration up to 5 mM. The protease was concentrated 1.6-fold with ammonium sulfate precipitation and dialysis. At least six protein bands were obtained from dialysate by electrophoresis. Four clear protein bands with caseinolytic activity were detected by zymography. Dialysate was further purified by anion-exchange chromatography and the caseinolytic active fraction showed a single band between 29 and 36 kDa of reducing conditions.
authors: Department of Biology, Faculty of Arts and Science, Hitit University , Corum , Turkey .
authors: Burçin
authors: B
authors: Özçelik
authors: Pınar
authors: P
authors: Aytar
authors: Serap
authors: S
authors: Gedikli
authors: Ezgi
authors: E
authors: Yardımcı
authors: Figen
authors: F
authors: Çalışkan
authors: Ahmet
authors: A
authors: Çabuk
mesh_terms: D001407:Bacillus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D010450:Endopeptidases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D004795:Enzyme Stability; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D012988:Soil Microbiology; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D012967:Sodium Dodecyl Sulfate; D010450:Endopeptidases; C087822:alkaline protease
keywords: Bacillus pumilus; hydrocarbon-contaminated soil; production; protease; purification