Journal of Proteomics & EnzymologyISSN: 2470-1289

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Research Article, J Proteomics Enzymol Vol: 6 Issue: 1

Partial Purification and Characterization of Cellulase (Ec.3.2.1.4) from Aspergillus niger Using Saw Dust as Carbon Source

Ekpa Emmanuel* and Adebisi Queen
Department of Biosciences, College of Natural and Applied Sciences, Salem University Lokoja, Nigeria
Corresponding author : Ekpa Emmanuel
Department of Biosciences, College of Natural and Applied Sciences, Salem University Lokoja, Nigeria
Tel: +2347036135834
E-mail: [email protected]
Received: August 28, 2016 Accepted: December 17, 2016 Published: January 11, 2017
Citation: Emmanuel E, Queen A (2017) Partial Purification and Characterization of Cellulase (Ec.3.2.1.4) from Aspergillus niger Using Saw Dust as Carbon Source. J Proteomics Enzymol 6:1. doi: 10.4172/2470-1289.1000128

 

Abstract

Cellulase (EC.3.2.1.4) was isolated and partially purified from the submerged culture of Aspergillus niger through a simple four step purification step using homogenization to obtain crude filtrate, ammonium sulphate precipitation, carboxymethyl cellulose chromatography, and sephadex G-200 chromatography. A purification fold of 1.00 and 1.26 were gotten for crude filtrate and ammonium sulphate precipitation respectively, while 0.56 and 0.41 were gotten for carboxymethyl cellulose and sephadex G-200. The highest yield of 96.4% was recorded for (NH4)2SO4 precipitation. Kinetic studies gave Km and Vmax of 0.01 mg/ml and 1.02 μmole/ml/min for carboxymethyl cellulose and 0.02 mg/ml and 0.96 μmole/ml/min for sephadex G-200. Optimum temperature and pH of 40°C and 8.0 were obtained for the cellulase. Ethanol was found to inhibit the activity of the enzyme higher than other inhibitors tested. The implication of this is that given a more suitable carbon source and a longer culturing time, a very useful cellulase with commercial value could be produced.

Keywords: Cellulases; Saw dust; Aspergillus niger; Enzyme purification

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