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CLONING AND EXPRESSION OF LACCASE GENE PRODUCED FROM Bacillus subtilis ZHR IN E. coli
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Ten isolates of Laccase producing bacteria were obtained from 50 isolates belongs to Bacillus sp. These isolates were isolate from different local areas of soil in baghdad.The isolates were subjected to primary screening , and revealed high production of enzyme based on consumed time during the reaction with syringaldazine ( SGZ ) reagent at 37 ° C. These isolates were subjected to secondary screening. The results revealed that isolate 136 was the best producers with highest enzyme activity of 47.0 U/ml. Identification of this isolate was achieved by studying morphological, microscopic characteristic , Vitek2 compact system and studying gene sequencing analysis of 16S rRNA gene .The results revealed that the isolate was belongs to Bacillus subtilis and identified as Bacillus subtilis ZHR. The laccase gene from Bacillus subtilis ZHR was extracted and amplified by PCR technique, then cloned in expressed Escherichia coli to produce biologically active enzyme with enzyme activity of 108.6 U/ml and specific activity 332.14 U/mg .
University of Baghdad - College of Agriculture
Title: CLONING AND EXPRESSION OF LACCASE GENE PRODUCED FROM Bacillus subtilis ZHR IN E. coli
Description:
Ten isolates of Laccase producing bacteria were obtained from 50 isolates belongs to Bacillus sp.
These isolates were isolate from different local areas of soil in baghdad.
The isolates were subjected to primary screening , and revealed high production of enzyme based on consumed time during the reaction with syringaldazine ( SGZ ) reagent at 37 ° C.
These isolates were subjected to secondary screening.
The results revealed that isolate 136 was the best producers with highest enzyme activity of 47.
0 U/ml.
Identification of this isolate was achieved by studying morphological, microscopic characteristic , Vitek2 compact system and studying gene sequencing analysis of 16S rRNA gene .
The results revealed that the isolate was belongs to Bacillus subtilis and identified as Bacillus subtilis ZHR.
The laccase gene from Bacillus subtilis ZHR was extracted and amplified by PCR technique, then cloned in expressed Escherichia coli to produce biologically active enzyme with enzyme activity of 108.
6 U/ml and specific activity 332.
14 U/mg .
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