Abstract

GENETIC ENHANCEMENT TO INCREASE THE PRODUCTION EFFICIENCY OF GLUTAMIC ACID FROM BACILLUS SUBTILIS EN3

Eman J. Al-Attar, Shaymaa H. Al-Rajhi* and Nidhal M. Salih

ABSTRACT

In an attempt enhance production of glutamic acid from wild bacteria B.subtilis EN3 by applying protoplast fusion technique by lysozyme and stimulate the protoplast fusion by using Polyethylene glycol and Ca++, we achieved fusant cells with 1.7×10-2 frequency when they grow on solid production medium. Among 30 fusant isolate the isolate EN3A1-P19 was selected as a best fusant for its excellence in glutamic acid production of which reached 13.7 mg/ml and the improved to 685% of what was produced by the parental cells. In other attempts to increase the production of glutamic acid wild bacteria under study EN3 and fusant EN3A1-P19 was exposed to UV to increase random mutations, its ability to produce glutamic acid has been detected and observed an increasing in the productivity of mutagens in glutamic acid which reaches 15 mg/ml and improved the percentage of productive to750% of what produce by wild isolation. While a superiority of fusant protoplast and mutagens was observed by UV light EN3A1-P19U7 from the rest of the isolates fusant and mutagens by irradiation UV, the glutamic acid production was reached 16.4 mg/ml.Glutamic acid was extracted from production medium by centrifugation (speed of 5000 for 20 minutes). Objective: Improve glutamic acid production by using protoplast fusion and UV mutagenesis.

Keywords: B. subtilis, glutamic acid, protoplast fusion, UV mutagenesis, random mutation.