the cytoplasmic and periplasmic expression levels and folding of organophosphorus hydrolase enzyme in escherichia coli

نویسندگان

ali mohammad latifi applied biotechnology research centre, baqiyatallah university of medical sciences, tehran, ir iran

khosro khajeh department of biochemistry, faculty of science, tarbiat modares university, tehran, ir iran

gholamreza farnoosh applied biotechnology research centre, baqiyatallah university of medical sciences, tehran, ir iran; applied biotechnology research centre, baqiyatallah university of medical sciences, tehran, ir iran. tel/fax: +98-9155437109

kazem hassanpour medical school, sabzevar university of medical sciences, sabzevar, ir iran

چکیده

conclusions the activities studied by the production of pnp were determined by following the increase at 410 nm. the maximum pnp was produced at 30°c with an optical density of 10.62 in the presence of cytoplasmic expression of oph (pet21a-opd). consequently, our results suggest cytoplasmic expression system as an appropriate candidate with a high amount of oph in spite of inclusion body formation, which needs an additional refolding step. results sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page) analysis showed a band of about 37 kda with a maximum expression level at 30°c from pet21a-opd.however, the obtained results of the periplasmic space extraction of oph (pet26b-opd) showed a very weak band, while the cytoplasmic expression of oph (pet21a-opd) produced a strong protein band. background organophosphorus hydrolase (oph) is a type of organophosphate-degrading enzyme which is widely used in the bioremediation process. objectives in this study, the periplasmic and cytoplasmic productions and the activity of recombinant oph in escherichia coli were investigated and compared using two pet systems (pet21a and pet26b). materials and methods the sequence encoding the opd gene was synthesized and expressed in the form of inclusion body using pet21a-opd and in the periplasmic space in pet26b-opd.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

The Cytoplasmic and Periplasmic Expression Levels and Folding of Organophosphorus Hydrolase Enzyme in Escherichia coli.

BACKGROUND Organophosphorus hydrolase (OPH) is a type of organophosphate-degrading enzyme which is widely used in the bioremediation process. OBJECTIVES In this study, the periplasmic and cytoplasmic productions and the activity of recombinant OPH in Escherichia coli were investigated and compared using two pET systems (pET21a and pET26b). MATERIALS AND METHODS The sequence encoding the opd...

متن کامل

Evaluation of the organophosphorus hydrolase enzyme activity in creams and investigation of its stability

The main purpose of this project is investigation of the organophosphorus hydrolase (OPH) enzyme activity in water in oil (w/o) and oil in water (o/w) creams and investigation of the OPH enzyme stability in formulated creams. OPH enzyme was extracted and purified from strain flavobacterium. The w/o and o/w creams were prepared using different formulations. In order to achieve an emulsion with m...

متن کامل

human tissue plasminogen activator expression in escherichia coli using cytoplasmic and periplasmic cumulative power

tissue plasminogen activator (tpa) is a serine protease, which is composed of five distinct structural domains with 17 disulfide bonds, representing a model of high-disulfide proteins in human body. one of the most important limitations for high yield heterologous protein production in escherichia coli (e. coli) is the expression of complex proteins with multiple disulfide bridges. in this stud...

متن کامل

: the effect of sericin levels (silk glue protein) on rate of in vitro maturation, fertilization and culture of sheep oocytes

هدف از آزمایش اول بررسی اثر سطوح مختلف سریسین [0 (control), 0.1, 0.5, 1.0, 2.5 %] افزوده شده به محیط , ivm بر cumulus cell expansion، بلوغ هسته و توسعه متوالی جنین، در گوسفندان نژاد سنجابی در فصل تولید مثلی می باشد. از سرگیری میوز به وسیله خارج شدن اولین پولار بادی اندازه گیری و هم چنین درصد رسیدن جنین های دو سلولی به مرحله کلیواژ و بلاستوسیت نیز به عنوان نشانه ای از میزان شایستگی توسعه اولیه ج...

Expression Cloning of Recombinant Escherichia coli lacZ Genes Encoding Cytoplasmic and Nuclear P-galactosidase Variants

Objective(s) Nonviral vector can be an attractive alternative to gene delivery in experimental study. In spite of some advantages in comparison with the viral vectors, there are still some limitations for efficiency of gene delivery in nonviral vectors. To determine the effective expression, the recombinant Escherichia coli lacZ genes were cloned into the different variants of pcDNA3.1 and the...

متن کامل

Cloning and optimization of phytase enzyme gene expression in Escherichia coli

Introduction Phytase is an enzyme that has the ability to break down phytic acid into myoinositol and mineral phosphate, and widely uses as an additive in animal foods. The aim of this study was to achieve a high level of bacterial phytase expression in PET26b expression host. Materials and Methods To generate the recombinant phytase enzyme, the target gene was introduced into the expression ...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
jundishapur journal of microbiology

جلد ۸، شماره ۱۲، صفحات ۰-۰

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023