Partial purification and characterization of α-amylase produced by Aspergillus oryzae using spent-brewing grains

Anil Kumar Patel, K. Madhavan Nampoothiri, Sumitra Ramachandran, George Szakacs, Ashok Pandey

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Solid-state fermentation (SSF) was carried out to produce α-amylase from Aspergillus oryzae (IFO 30103) using spent brewing grains (SBG) as substrate. A maximum of 11296 U/gds amylase activity was obtained after 48 h of fermentation. The extracted enzyme was subjected to partial purification by ammonium sulphate fractionation. Maximum specific activity was obtained with 40-70% fraction. SDS-PAGE of the corresponding sample revealed an approximate 66 kDa band, which was confirmed by activity staining as α-amylase. It was optimally active atpH 5 and 50°C by using 1% starch as substrate concentration. The partially purified α-amylase loses activity rapidly above 50°C but it can be retained in the presence of Ca2+. Presence of Mn2+ and Fe2+ enhanced the enzyme activity and is almost doubled in presence of Mn2+. However, in the presence of Hg2+ and Cu2+ the activity is reduced. Hg2+ reduced the enzyme activity by half.

Original languageEnglish
Pages (from-to)336-341
Number of pages6
JournalIndian Journal of Biotechnology
Volume4
Issue number3
Publication statusPublished - Jul 1 2005

    Fingerprint

Keywords

  • Aspergillus oryzae
  • Solid-state fermentation
  • Spent brewing grains
  • α-Amylase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Plant Science

Cite this