Estrogen circulating in blood has been proved to be a strong biomarker for breast cancer. A β-glucuronidase enzyme (GUS) from human gastrointestinal tract (GIT) microbiota including probiotics has significant involvement in enhancing the estrogen concentration in blood through deconjugation of glucuronidated estrogens. The present project has been designed to explore GIT microbiome-encoded GUS enzymes (GUSOME) repertoire in normal human and breast cancer patients. For this purpose, a total of nineteen GUS enzymes from human GIT microbes, i.e., seven from healthy and twelve from breast cancer patients have been focused on. Protein sequences of enzymes retrieved from UniProt database were subjected to ProtParam, CELLO2GO, SOPMA (secondary structure prediction method), PDBsum (Protein Database summaries), PHYRE2 (Protein Homology/AnalogY Recognition Engine), SAVES v6.0 (Structure Validation Server), MEME version 5.4.1 (Multiple Em for Motif Elicitation), Caver Web server v 1.1, Interproscan and Predicted Antigenic Peptides tool. Analysis revealed the number of amino acids, isoelectric point, extinction coefficient, instability index and aliphatic index of GUS enzymes in the range of 586–795, 4.91–8.92, 89980–155075, 25.88–40.93 and 71.01–88.10, respectively. Sub-cellular localization of enzyme was restricted to cytoplasm and inner-membrane in case of breast cancer patients’ bacteria as compared to periplasmic space, outer membrane and extracellular space in normal GIT bacteria. The 2-D structure analysis showed α helix, extended strand, β turn and random coil in the range of 27.42–22.66%, 22.04–25.91%, 5.39–8.30% and 41.75–47.70%, respectively. The druggability score was found to be 0.05–0.45 and 0.06–0.80 in normal and breast cancer patients GIT, respectively. The radius, length and curvature of catalytic sites were observed to be 1.1–2.8 Å, 1.4–15.9 Å and 0.65–1.4, respectively. Ten conserved protein motifs with p < 0.05 and width 25–50 were found. Antigenic propensity-associated sequences were 20–29. Present study findings hint about the use of the bacterial GUS enzymes against breast cancer tumors after modifications via site-directed mutagenesis of catalytic sites involved in the activation of estrogens and through destabilization of these enzymes.

Maize (Zea mays L.) holds significance importance in Pakistan in addition to being the highest yielding cereal crop in the world. But its yield has been severely affected due to the different environmental constraints including heat stress. In order to meet the growing food demand there is dire need to produce climate resilient varieties of maize. For this purpose we need to exploit the biochemical and molecular mechanisms involved in providing heat resistance to maize. In present study four genes were selected which were reportedly involved in response against heat stimulus, but were not structurally and functionally characterized. Therefore in-silico approach was used in order to investigate these hypothetical/candidate proteins. The phylogenetic investigation was conducted to find the evolutionary relationship. The 3D structures were elucidated in addition to the secondary structure prediction. Conserved domains and their interactions with other proteins were analyzed for the functional annotation. Our results showed that the protein sequences of maize are showing constant evolutionary change similar to their homologues. Predicted structures and domains of HPs are involved in conferring heat tolerance to maize crop. NP_001148903.1 is one of the HP; its interactions with other proteins along with its predicted domains, suggests that it is involved in stress response as most of its interacting partners are stress responsive proteins. This theoretical overview of heat tolerance proteins in maize will help researchers in understanding their predicted structure and function which will aide them in designing the genetically engineered varieties of heat resilient maize.