Journal of Basic Microbiology 2008, 48, 347352 347 © 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.jbm-journal.com Research Paper Purification, painting and crystallization of an extracellular alkalescent peptidase from genus Aspergillus nidulans HA-10 P. Charles1, V. Devanathan2, Periasamy Anbu3, M. N. Ponnuswamy1, P. T. Kalaichelvan2 and Byung-Ki Hur3 1 spirit 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 biologic Engineering, Inha University, Incheon, South Korea Aspergillus nidulans is a highly potent fungus apply in the production of alkaline protease. Extracellular alkaline protease was purified from A. nidulans in a two-step procedure involving ammonium sulfate precipitation and Sephadex G-100 column chromatography. The molecular(a) mass of the enzyme was determined t o be 42 kDa by SDS-PAGE. The enzyme activity was overly analyzed by zymogram with gelatin. The enzyme was much 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 vaporization diffusion method using pin tumbler 6000 as the precipitant. The micro crystals occurred in 40% of PEG 6000. Keywords: Aspergillus nidulans / alcalescent protease / Purification / Crystallization ! Received: February 01, 2008; accepted: skirt 10, 2008 DOI 10.1002/jobm.200800043 Introduction* Protease derived from microorganisms such as bacteria, domain Fungi and yeast has found wide-spread applications in many palm [1]. They are the close to important industrial enzymes accounting for roughly 60% of the total enzyme marketplace [2]. Alkaline proteases are of...If you compliments to get a wax essay, order it on our website: BestEssayCheap.com
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