© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.jbm-journal.com
Research Paper
Purification, characterization and crystallization
of an extracellular alkalescent protease
from genus Aspergillus nidulans HA-10
P. Charles1, V. Devanathan2, Periasamy Anbu3, M. N. Ponnuswamy1, P. T. Kalaichelvan2
and Byung-Ki Hur3
1 Centre of Advanced make in Crystallography and Biophysics, University of Madras, Guindy Campus,
Chennai, India
2 Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai, Tamil Nadu, India
3 Department of Biological Engineering, Inha University, Incheon, South Korea
Aspergillus nidulans is a highly potent fungus used in the production of alkaline protease.
Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving
ammonium sulphate precipitation and Sephadex G-100 column chromatography. The
molecular(a) mass of the enzyme was determined to be 42 kDa by SDS-PAGE. The enzyme activity
was overly analyzed by zymogram with gelatin. The enzyme was more stable over a wide range of
pH (610) and the temperatures up to 50 °C. It showed best enzyme activity at pH 8.0 and
a temperature of 35 °C.
The protease enzyme was completely inhibited by the serine protease
inhibitor of phenylmethylsulfonyl fluoride (PMSF). The crystallization of the purified enzyme
was performed by reprieve drop vapour diffusion method using pin tumbler 6000 as the precipitant.
The micro crystals occurred in 40% of PEG 6000.
Keywords: Aspergillus nidulans / Alkaline protease / Purification / Crystallization
Received: February 01, 2008; accepted: abut 10, 2008
DOI 10.1002/jobm.200800043
Introduction*
Protease derived from microorganisms such as bacteria,
kingdom Fungi and yeast has found wide-spread applications in
many fields [1]. They are the close to important industrial
enzymes accounting for about 60% of the total enzyme
marketplace [2]. Alkaline proteases are of...If you want to get a wax essay, order it on our website: Orderessay
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