Purification and characterization of α-amylase from Ganoderma tsuage growing in waste bread medium

  • Muhammad Irshad
  • Zahid Anwar
  • Muhammad Gulfraz
  • Hamama Islam Butt
  • Amir Ejaz
  • Haq Nawaz
Keywords: α-Amylase, purification, characterization, waste bread, Ganoderma tsuage

Abstract

The objective of this study was to purify and characterize the α-amylase for industrial perspective. The production of α-amylase through solid-state fermentation by Ganoderma tsuage was investigated by using waste bread as substrates. Production parameters were optimized as 2 mL of inoculum size, moisture 50%, additional carbon source (glucose) and nitrogen source (ammonium nitrate) 10:1, 1 mM/mL MgSO4, 0.75 mM/mL CaCl2 and 0.50 mM/mL KH2PO4. The purification value of α-amylase was observed as 1.2 fold with specific activity of 112 U/mg having a yield of 22%. Specific activity of α-amylase increased up to the level of 143 U/mg and had 1.5-fold purification factor having a yield of 6% after Sephadex gel filtration. Optimum value of α-amylase was obtained at 35°C and at pH 6 for the time duration of 72 h. The Km and Vmax values for α-amylase were 1.3 mg and 39 mg/min, respectively. Calcium chloride (CaCl2) was found to increase the activity of α-amylase while all other compounds seemed to have inhibitory action against α-amylase. Silver nitrate (AgNO3) was the strongest inhibitor and therefore would not be advised for use in future research against α-amylase production.

Keywords: α-Amylase, purification, characterization, waste bread, Ganoderma tsuage

Published
2015-12-16
Section
Articles

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eISSN: 1684-5315