Izvorni znanstveni članak
https://doi.org/10.15255/CABEQ.2016.831
Study on Some Properties of Calcium-dependent a-Amylase from Bacillus subtilis through Submerged Fermentation of Wheat Bran
M. Irfan
orcid.org/0000-0003-2955-4237
; Department of Biotechnology, University of Sargodha, Sargodha Pakistan
M. Nadeem
; Food & Biotechnology Research Center, Pakistan Council of Scientific & Industrial Research (PCSIR) Laboratories Complex, Ferozepur road Lahore, Pakistan 54600
Q. Syed
; Food & Biotechnology Research Center, Pakistan Council of Scientific & Industrial Research (PCSIR) Laboratories Complex, Ferozepur road Lahore, Pakistan 54600
H. Abdullah Shakir
; Microbial Biotechnology Laboratory, Department of Zoology, University of the Punjab new campus Lahore Pakistan
J. Iqbal Qazi
; Microbial Biotechnology Laboratory, Department of Zoology, University of the Punjab new campus Lahore Pakistan
Sažetak
This study reveals the production of α-amylase through Bacillus subtilis by submerged fermentation of wheat bran as a substrate. Various conditions were optimized and maximum production was observed at initial medium pH of 7.0, incubation temperature
of 35 °C with agitation speed of 140 rpm for 48 h of fermentation period. Peptone, ammonium sulphate and soluble starch favored enzyme production as nitrogen and carbon source, respectively. The crude α-amylase was purified 2.03-fold by column (sephadex G-100) chromatography having specific activity of 480.0 U mg–1. The purified enzyme had molecular mass of 54.18 kDa as determined by 12 % SDS-PAGE. The optimum pH and temperature of the enzyme was 8.5 and 60 °C, respectively. Ca2+ played an important role in the stability of the enzyme retaining relative activity of 87 % and 67 % at pH 9.5 and 80 °C, revealing the Ca2+ dependency respectively. The enzyme was highly stable towards surfactants and oxidizing agents. Kinetic study indicated that the enzyme exhibit Km and Vmax value of 4.4 mg mL–1 and 714.2 U mL–1 using soluble starch as a substrate respectively.
Ključne riječi
α-amylase; purification; characterization; Bacillus sp
Hrčak ID:
172289
URI
Datum izdavanja:
6.1.2017.
Posjeta: 1.322 *