Production of an alkaline protease using Bacillus pumilus D3 without inactivation by SDS, its characterization and purification

ÖZÇELİK B., AYTAR ÇELİK P., Gedikli S., Yardimci E., ÇALIŞKAN F., ÇABUK A.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY, cilt.29, sa.3, ss.388-396, 2014 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 29 Sayı: 3
  • Basım Tarihi: 2014
  • Doi Numarası: 10.3109/14756366.2013.788503
  • Dergi Adı: JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.388-396
  • Anahtar Kelimeler: Bacillus pumilus, hydrocarbon-contaminated soil, production, protease, purification, MICROBIAL PROPERTIES, SERINE-PROTEASES, FERMENTATION, PROTEINASE, STRAIN, GENES, SOIL
  • Eskişehir Osmangazi Üniversitesi Adresli: Evet

Özet

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 degrees 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 degrees C and 40 degrees 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.