Figure 2 - uploaded by Kai Griebenow
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Asparaginase enzymatic activity. (A) Asparaginase presence. Asparaginase reacts with the substrate (L-asparagine) to form the enzyme−substrate complex and generate the product (ammonia); then the presence of ammonia is determined by the addition of the aquarium test kit solutions that shows a change of color (orange to green). (B) BSA presence. BSA is mixed with the substrate, and no reaction takes place; then the aquarium test kit solutions are added, and in this case the color remains (orange) since no ammonia is produced. Chemical structures are generated from published data 15 and visualized and modified with PyMOL and ChemDraw.
Source publication
Determining the catalytic activity of an enzyme can be the perfect method for its identification, for example during purification procedures or for isolation purposes. Herein, we used a pharmaceutically relevant protein to bring the concept of enzymatic activity to the classroom. We designed a hands-on interactive activity in which a medically rele...
Context in source publication
Context 1
... activity consisted of a hands-on interactive experiment to show the enzymatic activity of asparaginase, 12,15 alternating with an animated slideshow to introduce basic concepts: bionanotech- nology, protein size and composition, asparaginase as an antileukemia drug, recombinant protein production (in general), and the asparaginase enzymatic activity (Supporting Information 2). Finally, we finished the presentation with a competition between the two proteins (asparaginase vs BSA) (Figure 2). Through this methodology, the students were motivated by the adaptation of a recreational chemistry strategy; they were entertained with an experiment in which scientific concepts were perceived as something else more than a "magical color change". ...
Citations
The purpose of this study was to map the research literature on Biochemistry education, covering the scientific production indexed on the Web of Science over the past 66 years. The open‐source Bibliometrix R‐package, an R‐tool, was used to carry out the bibliometric analysis. Our results describe (1) how many articles were published per year and what is the annual average growth rate; (2) which are the core journals, authors, and publications in the field; (3) which countries and funding agencies contribute most to the development of research in the area; (4) the leading collaborative research and co‐citation networks; (5) which articles were the most cited in the past 10 years; and (6) which are the trending topics in the field. Our main contribution is offering insights into the evolution of the field. Also, the use of a quantitative methodological design, which covers a large volume of publications, and could identify possible gaps in the area.
Protein PEGylation refers to the covalent binding of polyethylene glycol (PEG) moieties to a protein and has been increasingly applied, mainly in the biopharmaceutical industry. This technique may increase bioavailability and stability by decreasing immunogenicity and proteolytic attack. Given the industrial interest and the relevance of this technique, an example of a common PEGylation reaction is herein proposed, using lysozyme as a model protein. This class is divided into three parts: the first one is composed by the PEGylation reaction per se, the second one aims at the analytical quantification of the PEGylation yield, and the third aims to understand the effects of PEGylation on protein activity. Students should gain skills in comprehending this type of chemical reaction, calculating reaction yields, and realizing the benefits of protein PEGylation. This experimental class comprises two lab classes with the expected duration of 2 h each and is intended for students in the third and fourth years of courses such as pharmaceutical sciences, biochemistry, chemistry, biotechnology, and correlated areas.
A laboratory experiment introducing the concept of chemical bioconjugation of proteins to undergraduate students in a therapeutically relevant context was developed. Initially, students installed an aldehyde functionality into a protein via the oxidation of the N-terminal threonine residue of the cholera toxin subunit B (CTB) protein, which was followed by subsequent modification via hydrazone addition under mild conditions with a chromophore bearing a distinct UV–vis-absorption peak. Students determined the yield of the reaction to be ca. 11% by HPLC coupled to UV–vis spectroscopy and developed key skills such as preparation of stock solutions, chemical manipulation of proteins, and analysis via HPLC. The reported experiment can be readily adapted for use with other proteins and may contribute to enhancing constructive alignment in interdisciplinary degree programs at the chemistry–biology interface.
