Expression and Functional Characterization of Oryzacystatin II Protein from Oryza sativa L. Indica Rice and Its Potential Role as a Bioinsecticide Against Major Insects of Rice.

Rice (Oryza sativaL.) is a crucial staple food crop globally, facing significant challenges from various pests that affect crop productivity and quality. Conventional pesticide usage has limitations, necessitating the development of sustainable pest management strategies. This study focuses on the expression, purification, and functional characterization of Oryzacystatin II (OC-II), a protein derived from O. sativaL. Indica rice, with the intent to evaluate its potential as a bioinsecticide against rice pests. The OC-II gene was expressed and purified, and purification confirmed its molecular weight (∼12 kDa) and protein sequence through LC-MS/MS analysis and Western blotting. The IC50 value of OC-II was calculated as 0.06 µM, and the inhibition was identified as a competitive inhibition. The protein exhibited efficient control of both pests at the nymph and adult stages, with lower probing marks observed on treated plants. The inhibition of cathepsin B enzyme activity in insects further confirmed the bioactivity of the OC-II protein. Molecular docking and molecular dynamics simulations provided insights into the interaction between the OC-II protein and cathepsin enzymes reported in BPH and WBPH. Further investigations can focus on optimizing production methods and exploring the specificity and efficacy of the OC-II protein against other crop pests to enhance its practical applications.

Biological Activity of Cyclic Peptide Extracted from Sphaeranthus amaranthoides Using De Novo Sequencing Strategy by Mass Spectrometry for Cancer.

Though there are several advancements and developments in cancer therapy, the treatment remains challenging. In recent years, the antimicrobial peptides (AMPs) from traditional herbs are focused for identifying and developing potential anticancer molecules. In this study, AMPs are identified from Sphaeranthus amaranthoides, a natural medicinal herb widely used as a crucial immune stimulant in Indian medicine. A total of 86 peptide traces were identified using liquid-chromatography-electrospray-ionisation mass spectrometry (LC-ESI-MS). Among them, three peptides were sequenced using the manual de novo sequencing technique. The in-silico prediction revealed that SA923 is a cyclic peptide with C-N terminal interaction of the carbon atom of ASP7 with the nitrogen atom of GLU1 (1ELVFYRD7). Thus, SA923 is presented under the orbitides class of peptides, which lack the disulfide bonds for cyclization. In addition, SA923, steered with the physicochemical properties and support vector machine (SVM) algorithm mentioned for the segment, has the highest in silico anticancer potential. Further, the in vitro cytotoxicity assay revealed the peptide has anti-proliferative activity, and toxicity studies were demonstrated in Danio rerio (zebrafish) embryos.