Further Analysis of <i>Burkholderia pseudomallei</i> MF2 and Identification of Putative Dehalogenase Gene by PCR

Abstract

Halogenated organic compounds are extensively and widely used as pesticides, herbicides, and antibiotics that contribute to the pollution. This research was aimed to further analyze and characterize a bacterium that has the ability to utilize 2,2dichloropropionic acid (2,2-DCP) as a model to study dehalogenase enzyme production. Microscopic observation, biochemical tests and PCR technique were carried out in order to characterize the isolated bacterium. Strain MF2 showed its ability to grow on 10 mM 2,2-DCP liquid minimal medium with doubling time of 13 h with maximum chloride ion released of 19.8 molCl -/mL. The 16S rDNA analysis suggested that strain MF2 belongs to the genus Burkholderia. This was supported by the microscopic observation and biochemical tests. Dehalogenase gene was observed when using only primers dehIfor1 and dehIrev2 derived from group I deh PCR primer sequences, whereas no amplification using dhlB-314-forward and dhlB-637-reverse (group II dehalogenase) and haloacetate dehalogenase (H2-1157-forward and H2-1662-reverse) PCR primer sequences. The results suggested that, possibly, dehalogenase from MF2 was related to group I deh. In conclusion, strain MF2 showed the ability to utilize 2,2-DCP as sole source of carbon and energy. Further analysis revealed the MF2 strain consisted of dehalogenase gene that could be used for degradation of man-made halogenated compounds present in the environment. Using existing dehalogenase PCR primers, it was possible to amplify the dehalogenase genes sequence.

Identification and Characterization of a 2,2-Dichloropropionic Acid (2,2-DCP) Degrading Alkalotorelant Bacterium Strain BHS1 Isolated from Blue Lake, Turkey

“…The rate of chloride ion released was proportional to the growth of strain BHS1 in the 2,2-DCP minimal media. Similar observation was seen in other studies as reported earlier [42,43]. Chloride ions released into the growth medium was due to the hydrolysis of 2,2-DCP inside the cells resulted in chloride ion released outside the cells [42,43] [ 44,45].…”

“…Similar observation was seen in other studies as reported earlier [42,43]. Chloride ions released into the growth medium was due to the hydrolysis of 2,2-DCP inside the cells resulted in chloride ion released outside the cells [42,43] [ 44,45]. The product of 2,2-DCP dehalogenation resulted in pyruvate, that was essential for cell growth.…”

Antarktika: Yaşam Bilimleri ve Biyoteknoloji Araştırmalarının Gözden Geçirilmesi

“…Ayrıca, bazı mikroorganizmalar düşük sıcaklıklarda mezofilik enzimlere göre daha aktif ve kararlı olan ve böylece düşük sıcaklıklı endüstriyel işlemler için kullanılabilecek soğuk aktif veya psikrofilik enzimler de üretebilmektedir [103] . [110], bazik göller [111] ve tarım alanları [112] gibi birbirinden farklı lokasyonlardan izole edilerek biyoremidasyon için kullanılmıştır. Kutup çalışmaları ile birlikte kutup bölgelerinden potansiyel türler izole edilip, pestisitler gibi kirleticilere karşı biyolojik mücadele ile doğa daha temiz hale getirilebilmektedir [85].…”

Isolation and Characterisation of the Degradation Potential of 2,2-Dichloropropionate (2,2-Dcp) by Bacillus Amyloliquefaciens From Gebeng

“…These bacteria produce enzymes known as dehalogenase which catalyze the cleavage of carbon-halogen bonds in halogenated organic compounds thereby producing some environmental friendly products by a process known as dehalogenation [8], [9]. Some examples of aerobic degradative bacteria include; Pseudomonas, Bacillus, Micrococcus and Rhodococcus while anaerobic degradative bacteria are Syntrophus, Methanospirillum, Syntrophobacter.…”