Vol.:(0123456789) 1 3 Molecular Biology Reports https://doi.org/10.1007/s11033-019-04787-7 ORIGINAL ARTICLE Isolation and screening of extracellular anticancer enzymes from halophilic and halotolerant bacteria from diferent saline environments in Iran Mahdis Zolfaghar 1  · Mohammad Ali Amoozegar 1  · Khosro Khajeh 2  · Hamid Babavalian 3  · Hamid Tebyanian 4 Received: 2 January 2019 / Accepted: 27 March 2019 © Springer Nature B.V. 2019 Abstract It was confrmed that several enzymes have anti-cancer activity. The enzymes L-asparaginase, L-glutaminase, and L-arginase were chosen according to amino acids starvation in cancer cells and screened in halophilic and halotolerant bacteria, given probably less immunological reactions of halophilic or halotolerant enzymes in patients. Out of 110 halophilic and halotol- erant strains, isolated from diferent saline environments in Iran and screened, some could produce a variety of anticancer enzymes. A total of 29, 4, and 2 strains produced L-asparaginase, L-glutaminase, and L-arginase, respectively. According to the phenotypic characteristics and partial 16S rRNA gene sequence analysis, the positive strains—strains with the abil- ity to produce these anticancer enzymes—were identifed as the members of the genera: Bacillus, Dietzia, Halobacillus, Rhodococcus, Paenibacillus and Planococcus as Gram-positive bacteria and Pseudomonas, Marinobacter, Halomonas, Idiomarina, Vibrio and Stappia as Gram-negative bacteria. The production of anticancer enzymes was mostly observed in the rod-shaped Gram-negative isolates, particularly in the members of the genera Halomonas and Marinobacter. Most of the enzymes were produced in the stationary phase of growth and the maximum enzyme activity was experienced in strain GBPx3 (Vibrio sp.) for L-asparaginase at 1.0 IU/ml, strain R 2 S 25 (Rhodococcus sp.) for L-glutaminase at 0.6 IU/ml and strain GAAy3 (Planococcus sp.) for L-arginase at 3.1 IU/ml. The optimum temperature and pH for L-asparaginase and L-glutaminase activities in selected strains were similar to the physiological conditions of human body and the enzymes could tolerate NaCl up to 7.5% concentration. Keywords Anticancer enzymes · Halophiles · L-asparaginase · Halophilic · Halotolerant enzymes Introduction Cancer is characterized by the uncontrolled and invasive growth of cells, due to the failure occurred in the control mechanisms of the cell growth and proliferation cycles [1]. Although conventional anticancer therapies, such as surgi- cal resection, radiotherapy, and chemotherapy are efective in about half of the cancer cases, such non-specifc methods are inefective, presenting many undesirable side efects. Thus, the improvement or replacement of non-specifc and invasive conventional treatment methods are being consid- ered in cancer therapy researches, particularly the methods targeting the cancer cells specifcally [2]. Enzyme therapy is one of these methods leading to more efective and specifc treatment of cancer [3]. Using certain kinds of enzymes, capable of depleting cells from a certain type of amino acids, essential for the growth of malignant cells but not crucial for the normal cells survival, is the main idea of enzyme Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-019-04787-7) contains supplementary material, which is available to authorized users. * Mohammad Ali Amoozegar amoozegar@ut.ac.ir 1 Extremophiles Laboratory, Department of Microbiology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, P. O. Box 14155–6455, Tehran, Iran 2 Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran 3 Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran 4 Research Center for Prevention of Oral and Dental Diseases, Baqiyatallah University of Medical Sciences, Tehran, Iran