Isolation and identification of L-asparaginase producing bacteria from soil and effluent of slaughterhouse in Isfahan

Document Type : Research Article

Authors

• Fereshteh Hozoorbakhsh ¹
• Mozhgan Ghiasian ¹
• Fereshte ghandehari ¹
• Zarrindokht Emami-Karvani ¹
• Maryam Khademi Dehkordi ²

¹ Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran.

² Department of Biology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran.

10.22034/ijmcm.2022.693586

Abstract

L-asparaginase is one of the most important treatments for acute lymphoblastic leukemia. The aim of this study was isolation and molecularly identification for L-asparaginase producer bacteria. In this study, samples were taken from the effluent and soil of a slaughterhouse in Isfahan. M9 culture medium was used to screen the L-asparaginase producer bacteria. After screening, qualitative measurement of L-asparaginase was performed based on the color change in the media. The L-asparaginase producer bacteria were identified by biochemical and molecular methods. 114 colonies were isolated from the samples. 5 colonies were able to produce L-asparaginase. The halo diameter of L-asparaginase production in bacteria numbers SS18, SS19, SE64, SE112 and SE114 were 21, 22, 30, 33 and 55 mm, respectively. 16SrRNA sequences showed that strain SS18 was 99% likely to belong to Stenotrophomonas maltophilia, strain SS19 was 99% likely to belong to Chryseobacterium indologenes, strain SE64 was 99% likely to belong to the genus Chryseobacterium, strain SE112 was 99% likely to belongs to Bacillus safensis species and strain number SE114 probably 100% belongs to Bacillus velezensis species.  Since, L-asparaginase from different bacteria has shown different anti-cancer effects, finding the L-asparaginase producer microorganisms is one of the ways that lead us to have an enzyme with ideal medical aspects. One of the best environments for finding these bacteria is slaughterhouse soil and effluent, which are high in protein and can therefore be a potential source of bacteria that produce L-asparaginase.

Keywords

• L-Asparaginase
• Screening
• Slaughterhouse
• Phylogenetic tree
• PCR

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