Understanding the interaction between proteins and polyphenols is of significance to food industries. The aim of this research was to investigate the mode of aggregation for trypsin-EGCG (Epigallocatechin-3-gallate) complexes. For this, the complex was characterized by fluorescence spectroscopy, circular dichroism (CD) spectra, small-angel X-ray scattering (SAXS), and atomic force microscope (AFM) techniques. The results showed that the fluorescence intensity of trypsin-EGCG complexes decreased with increasing the concentration of EGCG, indicating that the interaction between trypsin and EGCG resulted in changes in the microenvironment around fluorescent amino acid residues. The results of CD analysis showed conformational changes in trypsin after binding with EGCG. The results from SAXS analysis showed that the addition of EGCG results in the formation of aggregates of trypsin-EGCG complexes, and increasing the concentration of EGCG resulted in larger aggregates. AFM images showed that the trypsin-EGCG complex formed aggregates of irregular ellipsoidal shapes with the size of about 200 × 400 × 200 nm, with EGCG interconnecting the trypsin particles. Overall, according to these results, it was concluded that the large aggregates of trypsin-EGCG complexes are formed from several small aggregates that are interconnected. The results of this study shed some light on the interaction between digestive enzymes and EGCG.
Superoxide dismutases (SODs) are a group of enzymes that have a crucial role in controlling oxidative stress in plants.Here, we synthesized an environmentally friendly SOD mimic, SODm-123, from L-aspartic acid and manganese oxide. SODm-123 showed similar enzymatic activity to Mn-SOD. To gain insights into the role of SODm-123 in oxidative stress tolerance, a series of experiments were conducted to assess the physiological and molecular responses of tomato plants when treated with SODm-123. The results showed that the levels of O 2 −• and H 2 O 2 in tomato cells were affected by SODm-123 treatment, indicating that SODm-123 can control oxidative stress like Mn-SOD. The results also exhibited that SODm-123 increased the contents of photosynthetic pigments. However, it was noted that SODm-123 resulted in a reduction in the content of soluble sugar and MDA. These results indicate that SODm-123 promoted the efficiency of photosynthesis by regulating the content of H 2 O 2 . To further investigate the role of SODm-123 in controlling oxidative stress, a transcriptome analysis was used to identify differentially expressed genes (DEGs) associated with SODm-123 treatment. The results indicated that SODm-123 treatment resulted in 341 differentially expressed genes (DEGs) in treated tomato leaves at 96 h after treatment. Kyoto encyclopedia of genes and genomes (KEGG) revealed that DEGs were involved in pathways such as photosynthetic pigment biosynthesis, ABC transporters, sugar metabolism, and MAPK signaling, which further confirmed a positive role of SODm-123 in improving stress tolerance in plants. Overall, the results of this study suggest that SODm-123 promotes the growth and development of tomato seedlings and therefore can be used as a potential growthpromoting agent for plants.
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