Cloning, expression, and enzymatic activity evaluation of cholesterol oxidase gene isolated from a native Rhodococcus sp
Journal Title: International Journal of Medical Research & Health Sciences (IJMRHS) - Year 2016, Vol 5, Issue 10
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
- EP ID EP12100
- DOI -
- Views 267
- Downloads 11
Traditional herbal medicine for weight management: A Review
The prevalence of obesity is increasing at an alarming rate in recent years. Many drugs for weight managing are available in the market; however their adverse effects and hazards have not been thoroughly evaluated, ther...
Respiratory Manifestation of Malaria: An Update
Globally, malaria is the most important parasitic disease, representing a public health problem in more than 90 countries. In recent years, there has been an increased incidence of respiratory complications. Acute respi...
SOURCE OF STRESSORS AND EMOTIONAL DISTURBANCES AMONG UNDERGRADUATE SCIENCE STUDENTS IN MALAYSIA
Introduction: Higher education is considered as a stressful period in students’ life which they have to cope with since they are facing a variety of demands such as living away from their families, a heavily loaded cu...
The Effect of Logo Therapy on Life Orientation of Cancer Patient
The aim of the present research is to determine the effect of logo therapy on life orientation of cancer patients. The research method was quasi-experimental with control group and testing. Statistical population includ...
Osteopathic manipulative treatment results in sustained relief from spinal pain in older patients: A pilot crossover study
Osteopathic Manipulative Treatment is commonly used to complement conventional treatment of back pain. The present study verified whether OMT, associated with training program, reduces spinal pain in older. A pilot ran...