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Title: Partial purification and characterization of -amylase produced by Aspergillus oryzae using spent-brewing grains
Authors: Patel, Anil Kumar
Nampoothiri, K Madhavan
Ramachandran, Sumitra
Szakacs, George
Pandey, Ashok
Keywords:  -Amylase
Aspergillus oryzae
solid-state fermentation
spent brewing grains
Issue Date: Jul-2005
Publisher: CSIR
IPC CodeInt. Cl.7 A01N63/02; C12N9/30; C12R1: 69
Abstract: Solid-state fermentation (SSF) was carried out to produce -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 -amylase. It was optimally active at pH 5 and 50°C by using 1% starch as substrate concentration. The partially purified -amylase loses activity rapidly above 50oC 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.
Page(s): 336-341
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Source:IJBT Vol.04(3) [July 2005]

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