Multi-functional enzymes are one of the nature's solutions to facilitate metabolic pathways, thus several reactions are regulated and performed simultaneously on one polypeptide chain. Inspired by nature, artificial chimeric proteins have been designed to reduce the production costs and improve the performance. One of the interesting applications of this method is in the plant-based industries such as feed additive, waste treatment, biofuel production, and pulp and paper bleaching. In fact, the heterogeneous texture of plants needs using a combination of different enzymes to achieve an optimal quality in the manufacturing process. Given that xylans are the most abundant non-cellulosic polysaccharides in nature, xylanases are widely utilized in the mentioned industries. In this regard, several studies have been conducted to develop the relevant chimeric enzymes. Despite the successes that have been attained in this field, misfolding, functional or structural interference, and linker breakage have been reported in some cases. The present paper reviews the research to introduce the prerequisites to design an appropriate chimeric xylanase.
Rasburicase is an expensive treatment used to control hyperuricemia caused by tumour lysis syndrome (TLS). In this study, a non-chromatographic method was designed based on nano-oil bodies for convenient and economical purification of the recombinant uricase. For this purpose, two chimaeras were synthesized with a different arrangement of the uricase, caleosin and intein fragments. After confirming the protein expression by measuring the uricase activity at 293 nm, purification was conducted through oil-body construction. The results were resolved on the 12% SDS-PAGE gel. Finally, the stability of the oil bodies was examined against different salts, surfactants, temperatures, and pH values. According to our results, the overexpression of uricase–caleosin chimaera under the T7 promoter in Escherichia coli led to the production of soluble protein, which was successfully purified by artificial oil bodies. The active uricase was subsequently released through the self-splicing of intein. Further investigations highlighted the importance of the free C-terminus of caleosin in constructing artificial oil bodies. Moreover, surfactants and low temperature, in contrast to salts, improved the stability of oil bodies. In conclusion, caleosins are an efficient purification tag reducing the cost of purification compared to conventional chromatography methods. Graphical Abstract
Plant cells store energy in oil bodies constructed by structural proteins such as oleosins and caleosins. Although oil bodies usually accumulate in the seed and pollen of plants, caleosins are present in various organs and organelles. This issue, coupled with the diverse activities of caleosins, complicates the description of these oleo-proteins. Therefore, the current article proposes a new classification based on the bioinformatics analysis of the transmembrane topology of caleosins. Accordingly, the non-membrane class are the most abundant and diverse caleosins, especially in lower plants. Comparing the results with other reports suggests a stress response capacity for these caleosins. However, other classes play a more specific role in germination and pollination. A phylogenetic study also revealed two main clades that were significantly different in terms of caleosin type, expression profile, molecular weight, and isoelectric point (P < 0.01). In addition to the biochemical significance of the findings, predicting the structure of caleosins is necessary for constructing oil bodies used in the food and pharmaceutical industries.
Deaths from tuberculosis have long gripped people and threatened human health. The need for new drug compounds are critically sensed by medical scientists and practitioners due to the emergence of new strains and the slow rate of discovering novel medicines for this disease. Since plants are a rich source of diverse drug compounds, they are among the best choices to achieve new ones. The study of all plants or their compounds is an almost impossible scenario; hence bio/cheminformatics methodology can be used to reduce time and cost spent in drug discovery. For this purpose, we made several databases of anti-mycobacterial plant compounds and further found filter criteria which were able to describe more predicted bioactive compounds by the established algorithm. Also, we present the survey of the developed resource by using bio/cheminformatics tools. The presence of several anti-mycobacterial compounds in the predicted algorithm and introduction of new active compounds represent the high potential of this method. In addition, the general profile of such bioactive molecules is pinpointed using molecular descriptors and cheminformatics approach.
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