Enzymatically active Delta(5)-3-ketosteroid isomerase (KSI) protein with a C-terminus his(6)-tag was produced following insoluble expression using Escherichia coli. A simple, integrated process was used to extract and purify the target protein. Chemical extraction was shown to be as effective as homogenization at releasing the inclusion body proteins from the bacterial cells, with complete release taking less than 20 min. An expanded bed adsorption (EBA) column utilizing immobilized metal affinity chromatography (IMAC) was then used to purify the denatured KSI-(His(6)) protein directly from the chemical extract. This integrated process greatly simplifies the recovery and purification of inclusion body proteins by removing the need for mechanical cell disruption, repeated inclusion body centrifugation, and difficult clarification operations. The integrated chemical extraction and EBA process achieved a very high purity (99%) and recovery (89%) of the KSI-(His(6)), with efficient utilization of the adsorbent matrix (9.74 mg KSI-(His(6))/mL adsorbent). Following purification the protein was refolded by dilution to obtain the biologically active protein. Seventy-nine percent of the expressed KSI-(His(6)) protein was recovered as enzymatically active protein with the described extraction, purification, and refolding process. In addition to demonstrating the operation of this intensified inclusion body process, a plate-based concentration assay detecting KSI-(His(6)) is validated. The intensified process in this work requires minimal optimization for recovering novel his-tagged proteins, and further improves the economic advantage of E. coli as a host organism.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1002/bit.21043 | DOI Listing |
Heart Rhythm O2
December 2024
Department of Internal Medicine, Burnett School of Medicine at Texas Christian University (TCU) and Consultants in Cardiovascular Medicine and Science, Fort Worth, Texas.
Background: The adoption of leadless pacemakers (LPMs) is increasing, yet the impact of body mass index (BMI) on procedural outcomes remains underexplored.
Objective: The purpose of this study was to explore the impact of BMI on in-hospital outcomes for patients receiving LPM implantation.
Methods: Data from the National Inpatient Sample from 2018-2021 were analyzed for patients older than 18 years who underwent LPM implantation, with specific inclusion and exclusion criteria applied.
JSLS
January 2025
Western New York Urology Associates, Cheektowaga, New York, USA. (Dr. Eddib).
Background: Sacrocolpopexy has become a favored treatment of pelvic organ prolapse due to its durability and efficacy. Sacrocolpopexy has not been standardized and there is no categorization scheme to facilitate communication or research efforts for the procedure. A systematic review was conducted to facilitate construction of a classification system for sacrocolpopexy based on extent of vaginal dissection described in the medical literature.
View Article and Find Full Text PDFIndian J Radiol Imaging
January 2025
Department of Neurology, Division of Cerebrovascular Medicine and Neurology, National Cerebral and Cardiovascular Center (NCVC), Suita, Osaka, Japan.
Physician-scientists serve as conduits between clinical practice and scientific research, leveraging their unique expertise to improve patient care and drive medical innovation. This article highlights the indispensable role of research and publishing in promoting evidence-based practices, facilitating professional growth, and shaping public health policy. Drawing on historical and contemporary examples, I examine the challenges faced by physician-scientists, such as ethical dilemmas and declining engagement in research, particularly in resource-constrained settings.
View Article and Find Full Text PDFJ Adv Pract Oncol
September 2024
From Clemson University, Clemson, South Carolina.
Molecular profiling (MP), which involves testing tissue, blood, or other body fluids to identify biomarkers, has become increasingly important in cancer treatment. Genomic tumor sequencing, a specific type of MP, is commonly used to identify specific gene variants or proteins that can be targeted for treatment. Germline testing is also routinely recommended for certain cancers.
View Article and Find Full Text PDFTransl Anim Sci
December 2024
Department of Agricultural and Environmental Sciences, College of Agriculture, Environment and Nutrition Sciences, Tuskegee University, Tuskegee, AL 36088, USA.
Hempseed meal (HSM) is a potential alternative feedstuff for livestock due to its high protein content, but it has not been approved for animal feed in the United States due to safety concerns. This study was conducted to determine the effects of HSM on feed intake, growth performance, serum biochemistry, ruminal papillae morphology, ruminal fermentation profiles, and carcass characteristics of intact male goats. Thirty-six Boer × Spanish intact male goats were randomly assigned to one of four experimental diets ( = 9 goats/diet): 0%, 10%, 20%, and 30% HSM on as-fed basis.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!