Optimal Conditions for Extraction and Purification of Penicillinase Enzyme
نویسندگان
چکیده مقاله:
Background and Objective: The present study aimed to evaluate the optimum conditions for extracting and purifying penicillinase enzyme. The enzyme source was Bacillus licheniformis 6346 obtained from the Iranian Research Organization for Science and Technology. Materials and Methods: B. licheniformis was cultured in a medium containing mineral salts, nitrogen compounds, and cultured carbon sources. Following the harvesting and preparation of bacterial suspension, the destruction of the bacterial cell wall and crude cell extraction was completed using a combination of lysozyme and weak ultrasonication. Afterward, enzyme amount was determined utilizing chromatography and electrophoresis. Furthermore, the optimum growth temperature and incubation time were assessed based on the absorbance of samples at 240 nm, enzyme activity, aerification intensity optimization, enzyme production time, and optimum pH. Results: Approximately 8 g of cells were retrieved in one liter of culture. The optimum pH was determined as 6.8. The heat stability evaluation of the enzyme at different times revealed that the enzyme was stable for more than 1 h at 20°C-45°C, while it is inactivated in 10 min at 60°C. Moreover, the best incubation time for this bacterium was obtained as 9 h at 30°C. In terms of aerification intensity, the highest enzyme production rate of 2394 U/mL was achieved in a 250 mL Erlenmeyer flask with a 50 mL culture medium at 200 RPM. More aerification led to the inhibitory effect of oxygen and reduced enzyme activity. Conclusion: Our findings showed that the bacterium B. licheniformis 6346 can be used for producing penicillinase enzyme in optimum conditions.
منابع مشابه
ENZYME-LINKED IMMUNOSORBENT ASSAY OF NEOPTERIN USING PENICILLINASE AS LABEL
An enzyme-linked immunosorbent assay for neopterin using penicillinase as marker enzyme is reported here by polyclonal antibodies against neopterin conjugated to bovine serum albumin which were raised in rabbits. Immunoglobulin fractions were purified and coated on wells of microtiter plates. A chain heterology was introduced in neopterin derivative and conjugated to penicillinase. The assa...
متن کاملDEVELOPMENT OF A SIMPLE AND SENSITIVE ENZYME- LINKED IMMUNOSORBENT ASSAY (ELISA) FOR CLINICAL MEASUREMENT OF TESTOSTERONE USING PENICILLINASE AS LABEL
An enzyme-linked immunosorbent assay using a homologous combination of antiserum raised against testosterone-3-0-carboxymethyloxime-bovine serum albumin (T-3-0-CMO-BSA ) and penicillinase-labelled T-3-0-CMO was developed. This assay was utilized to measure testosterone in serum samples of male and female subjects. The sensitivity of the assay is 50pg/well and the antibody developed crossrea...
متن کاملinvestigation of extraction and purification of histone acetyl transferase enzyme from saccharomyces cerevisiae and assay enzyme activity
متن کامل
extraction and acetylation of purified trypsin from bovin pancreas and study of some its physico-chemical properties
آنزیم تریپسین در شرایط قلیایی ناپایدار می باشد .و فعالیت پروتئولیتیکی تریپسین منجربه خود هضمی آن در جایگاههای خاصی می گردد. بنابر این آنزیمی با ناپایداری بالا محسوب میگردد. در سالهای اخیر موفق شدند که با ایجاد تغیرات شیمیایی با اضافه کردن فلزات خاص ، کلسیم و یا عمل استیلاسیون منجر به افزایش پایداری آنزیم تریپسین گردند. مطالعات در حال حاضر نشان می دهد که تریپسین استیله شده فعالیت آنزیمی خود را ...
15 صفحه اولa benchmarking approach to optimal asset allocation for insurers and pension funds
uncertainty in the financial market will be driven by underlying brownian motions, while the assets are assumed to be general stochastic processes adapted to the filtration of the brownian motions. the goal of this study is to calculate the accumulated wealth in order to optimize the expected terminal value using a suitable utility function. this thesis introduced the lim-wong’s benchmark fun...
15 صفحه اولa study on thermodynamic models for simulation of 1,3 butadiene purification columns
attempts have been made to study the thermodynamic behavior of 1,3 butadiene purification columns with the aim of retrofitting those columns to more energy efficient separation schemes. 1,3 butadiene is purified in two columns in series through being separated from methyl acetylene and 1,2 butadiene in the first and second column respectively. comparisons have been made among different therm...
منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ذخیره در منابع من قبلا به منابع من ذحیره شده{@ msg_add @}
عنوان ژورنال
دوره 15 شماره None
صفحات 684- 691
تاریخ انتشار 2021-12
با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023