Optimum conditions for isoamylase production by Pseudomonas sp.

Hameed M Jasim,Haneen S Abdul-Wahab

doi:10.24126/jobrc.2013.7.1.232

Abstract

Different nutritional and cultural factors were studied to determine the optimum conditions for isoamylase production by Pseudomonas sp. in a batch culture of the production medium. These factors include carbon, nitrogen and phosphate sources and their concentrations, temperature and pH. Results showed that the optimum conditions for isoamylase production by Pseudomonas sp. were achieved when the production medium was supplemented with maltose 1%, peptone 0.4%, and K2HPO4 0.4% as a carbon, nitrogen, and phosphate sources respectively, at initial medium pH 6, and incubation at 28°C for 24 hours. Under these conditions isoamylase productivity reaches the maximum, at which enzyme specific activity was 0.85 U/mg proteins.

Highlights

The use of enzymes is preferred as it offers a number of advantages including improved yields and favourable economics [1,2]
Effect of carbon source Six carbon sources (Lactose, sucrose, fructose, galactose, glucose and maltose) were used as a sole sources for carbon and energy to select the optimum for isoamylase production by Pseudomonas sp
Results mentioned in Figure (1) showed that the maximum production of isoamylase was obtained when the production medium was supplemented with maltose as a soul source for carbon and energy

Summary

Introduction

The use of enzymes is preferred as it offers a number of advantages including improved yields and favourable economics [1,2]. The main objective of this study is to determine the optimum conditions for enzyme production by a local isolate of Pseudomonas sp. Optimization of Isoamylase production The optimization experiments were carried out aerobically under batch cultivation conditions in the production medium. Fifty ml of this medium was distributed in 250 ml Erlenmeyer flasks inoculated with 1% of mid-exponential phase culture of the Pseudomonas sp., and incubated with shaking at 150rpm in a shaker incubator at 28C°. Enzyme assay Assay of isoamylase was achieved according to [17], by adding 1ml of the crude enzyme to the reaction mixture containing 5ml of 1% soluble glutinous rice starch solution, 1ml of 0.5M acetate buffer pH 3.5. Protein Concentration Protein concentration in culture medium was determined according to [19] by using Coomassie blue G-250 and Bovine serum albumin standard solution

Results and Discussion

Nitrogen Source

Phosphate Source