Cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp

Abstract

Cholesterol oxidase (CHO) is one of the valuable enzymes that play an important role in: measurement of serum cholesterol, food industry as a biocatalyst and agriculture as a biological larvicide. This enzyme was produced by several bacterial strains. Wild type enzyme produced by Rhodococcus sp. secret two forms of CHO enzyme: extra cellular and membrane bound type which its amount is low and unstable. The goal of the study was cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp. CHO gene was isolated from native bacteria and cloned into pET23a. In the next step, the construct was expressed in E.coli BL21 and induced by different concentration of IPTG ranges from 0.1 - 0.9 mM. This gene contains 1642 bp and encodes a protein consists of 533 amino acids. It has about 96 % homology with CHO gene isolated from Rhodococcus equi. The high expression was obtained in 0.5 mM concentration of IPTG after 4 hour induction. This recombinant enzyme had a molecular weight of 55 kDa, that secretion of intra cellular type is much more than extra cellular form. The optimum pH and temperature conditions for the recombinant enzyme were 7.5 and 45°C, respectively. CHO enzyme obtained from Rhodococcus sp. is a cheap enzyme with medical and industrial applications that can be produced easily and purified in large scale with simple methods.

Authors and Affiliations

Hamed Esmaeil Lashgarian| Hepatitis Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran, Corresponding E-mail: hamedesmaiili@gmail.com, Kiana Shahzamani| Dept. of Biology, Faculty of Basic Sciences, Lorestan University, Khorramabad, Iran, Sareh Jahanbakhsh| Research Center for Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran

Keywords

Related Articles

HISTOLOGICAL AND HISTOMETRIC STUDY OF TESTIS IN ALBINO RATS TREATED WITH AMLODIPINE

Amlodipine is the most common drug of choice to treat hypertension, one of its side effects is infertility and its effect on the testis of male albino rats is not well documented. Aim: To observe the effect of amlodipin...

CHECKING AND CORRECTING COMPLIANCE: FOCUSING RENAL PATIENTS

Compliance is a behaviour resulting from a specific set of cues and consequences. It is a self-care behaviour which entails obedience to a directive whereas noncompliance is self-care deficit, which calls for rejection...

The Relationship Between Leadership Styles and Personality Traits of Nursery Managers of Hospitals Affliated to Tehran Medical Sciences Universities

Background: Nursing and health care, will be challenged to respond the changing needs of people and changing nature society and this requires that nurses are in managerial roles until nurses needs management and support...

Comparison of the effects of Mycocin vaginal cream and Metronidazole vaginal gel on treatment of bacterial vaginosis: A randomized clinical trial

Metronidazole is recommended as the first-line treatment for Bacterial Vaginosis (BV). With respect to various side effects of metronidazole and increased antibiotic resistance, it seems necessary to find a medication w...

Novel Electron Spin Resonance-Enzyme Immunosorbent Assay for Detecting Occult Hepatitis B Infection in HCV Chronic Liver Disease

Background: Hepatitis B virus infection in patients who lack detectable hepatitis B surface antigen (HBsAg) is called occult hepatitis B infection (OHB). The very low level of HBV genome may hamper its detection by mole...

Download PDF file
  • EP ID EP12100
  • DOI -
  • Views 268
  • Downloads 11

How To Cite

Hamed Esmaeil Lashgarian, Kiana Shahzamani, Sareh Jahanbakhsh (2016). Cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp. International Journal of Medical Research & Health Sciences (IJMRHS), 5(10), 91-96. https://europub.co.uk/articles/-A-12100