Iranian Biomedical Journal
مجله بیومدیکال ایران
IBJ
Basic Sciences
http://ibj.pasteur.ac.ir
1
admin
1028-852X
2008-823X
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10.61186/ibj
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8888
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en
jalali
1388
10
1
gregorian
2010
1
1
14
1
online
1
fulltext
en
Extracellular Cholesterol Oxidase from Rhodococcus sp.: Isolation and Molecular Characterization
Extracellular Cholesterol Oxidase from Rhodococcus sp.: Isolation and Molecular Characterization
Related Fields
Related Fields
مقاله کامل
Full Length/Original Article
Background: Cholesterol oxidase (CHO) has various clinical and industrial applications. Recently, microbial CHO have received a great attention for their wide usage in medicine. Here, taxonomic characterizations of isolated strain from soil, optimization of the conditions for CHO production and biochemical characterizations of produced CHO enzyme were described. Finally, CHO gene was cloned into a cloning vector. Methods: Various samples were collected and cultivated in a screening medium consisting of cholesterol. For isolation of CHO-producing bacteria, well-grown colonies were inoculated into an optimized medium. Different biochemical and microbiological tests were performed on isolated bacteria to identify their properties. For phylogenic analysis, a partial sequence of l6s rRNA was amplified by PCR using universally conserved primers. A modified method was applied for determination of CHO activity. Then, extracellular CHO activity was assessed under different temperature, pH and cholesterol concentration conditions. Finally, CHO gene was amplified by PCR and cloned into STV28. Results: According to the morphological, cultural and biochemical tests, the isolated bacterium was identified as Rhodococcus sp. strain 501 and deposited in GenBank with accession number FN298676. Results showed that optimum temperature and pH for CHO activity were 35°C and 7.5, respectively. Alignment of nucleotide sequence of CHO gene showed 99% homology with other bacterial CHO genes. Conclusion: Rhodococcus sp. strain 501 produced significant levels of extracellular CHO in an optimized medium for a short period. CHO gene was cloned into cloning vector that can be a valuable tool for better identification and further studies on gene expression.
Rhodococcus, Cholesterol oxidase, Soil
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http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-170&slc_lang=en&sid=1
Hamed
Lashkarian
حامد
لشکریان
100319475328460056801
100319475328460056801
No
Jamshid
Raheb
جمشید
راهب
jam@nigeb.ac.ir
100319475328460056802
100319475328460056802
Yes
Kiana
Shahzamani
کیانا
شاه زمانی
100319475328460056803
100319475328460056803
No
Hossein
Shahbani
حسین
شاهبانی
100319475328460056804
100319475328460056804
No
Mehdi
Shamsara
مهدی
شمس آرا
100319475328460056805
100319475328460056805
No