Isolation, molecular identification and production investigation of Bacillus-producing L-Asparaginase from Jiroft microflora

Subject Areas : Microbial Biotechnology

Arastoo Badoei-dalfard ¹ (Associate Professors, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.)
Zahra Karami ² (Assistant Professors, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.)
Narjes Ramezani-pour ³ (M.Sc., Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.)

Received: 2016-03-04 Accepted : 2016-04-25 Published : 2016-11-01

Keywords: Screening, Bacillus, L-asparaginase,

Abstract :

Background & Objectives: Cancer cells are not capable of producing L-asparagine, and mainly are depended to the L-asparagine from the circulating plasma pools. L-Asparaginase (L-asparagine amidohydrolase EC 3.5.1.1) catalysis the conversion of L-asparagine to L-aspartate and ammonium. L-asparaginase is the first enzyme with anti-leukemic activity to be intensively studied in human beings. Isolation and identification of L-asparaginase producing bacteria from Jiroft microflora was the aim of this study. Materials & Methods: In this sectional study, soil and water samples were collected from citrus orchards and waste of milk factory in Jiroft, Kerman province. The bacterial-producing L-asparaginase was screened on agar medium supplied with L-asparagine and phenol red indicator dye. L-asparaginase activity was detected on the basis of pink color around the colony. The best strains were finally identified using PCR method. Results: 16S rRNA sequencing results showed that the best isolate-producing L-asparaginase belonged to the Bacillus genus. DJK23 strain produced the maximum L-asparaginase, approximately 165 U/ml. The our analysis showed that L-asparaginase was active in a pH rang of 5 to 8, with maximal enzyme activity at pH 7. It is mentioned that this enzyme retains 36% of its initial activity at pH 5. Conclusion: Although L-asparaginase from bacteria has been extensively characterized, a similar attention has not been paid to Bacillus. These results showed that Bacillus L-Asparaginase has desirable activity in the presence of neutral and acidic pH, which can be considered as a therapeutic and industrial enzyme.

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