In the present research work, the phytochemical and biological evaluation of Crude Methanol Extract (CME) and fractions of aerial parts of Berberis lycium Royle (family: Berberidaceae) was conducted. The results of phytochemicals analysis showed the presence of tannins, alkaloids and sterols. The n-hexane fraction presented significant activity against S. aureus (80%) and low activity against B. pumilus (34%), S. typhi (33%), K. pneumoniae (26%), S. pneumoniae (25%) and S. epidermidis (10%). The CHCl3 fraction showed significant results against S. aureus (80%) while low activity was exhibited against S. pneumoniae (32%) and B. pumilus (30%). The EtOAc fraction showed a low activity against B. pumilus (39%), S. typhi (33%), and E. aerogenes (24%). The aqueous fraction of the plant was found to have good and moderate activities. The CME showed low activity against K. pneumoniae (32%). The CME possesses low activity against T. hazrianum (10%), A. niger (10%), A. flavus (10%) and R. stolonifer (6%). The n-hexane, CHCl3, EtOAc and aqueous fraction displayed low activity against the test fungal pathogens. The CME showed 10% and 5% regulation of growth at 1000 and 100 µg/mL, respectively against L. minor. The n- hexane fraction showed 20% growth regulation at 1000 µg/mL, while a growth regulation of 10% was observed at 100 µg/mL. In the same way, CHCl3, EtOAc and aqueous fraction showed low growth regulation at 1000 and 100 µg/mL. No phytotoxic activity was observed at 10 µg/mL in any of the test samples. Significant activity on insects was observed for n-hexane fraction against Tribolium castaneum (80%) while the CHCl3 fraction reported a significant vi antitermite activity. Low cytotoxic effect was observed for aqueous (32.5%) and EtOAc (35%) against Artemia salina. At 1.6 mg/mL, the activities of CME, EtOAc and n-hexane were 54.50, 53.50 and 52.22%, respectively, whereas CHCl3 and aqueous fraction showed 49.70 and 46.49% activity at the same concentration. Different in vivo studies were carried out which revealed that various metabolites of this plant have potentially high analgesic and antidepressant effects. The results showed that CME is safe upto dose of 600 mg/ kg. Charcoal meal test was used to study gastrointestinal (GIT) motility induced by CME. The mean charcoal movement after treatment with CME was 23.41, 19.63, and 11.23, at doses 100,200 and 500mg/ kg body weight. The extract causes relaxation of the small intestine and can be used in the treatment of diarrhea and abdominal spasms. The results were dose-dependent, i.e., the movement was reduced upon increasing the dose. The results of the current study revealed that increase in the dose of the CME enhanced the antidepressant effect, implying that CME possesses antidepressant activity, using Forced Swimming Test (FST) in animals. The CME was tested for locomotor activity and results were recorded. The total lines crossed by mice in the wooden apparatus were 38, 26, and 17 at a dose of 500, 700 and 1000 mg/kg body weight, respectively. Allopathy is a biological action by which one or more biochemical substances are secreted by an organism to effect the growth, existence and reproduction of other species. The germination of T. aestivum seeds was highly inhibited by B. lycium. On the basis of polarity, four compounds were isolated from n-hexane fraction through column chromatography. Four known compounds namely nonacosan–1-ol (2), heptacosan–1-ol, β-sitosterol and oxyberberine were isolated from B. lycium. vii Further the Oxyberberine was subjected to various enzyme inhibitory assays like urease activity, α-Chymotripson, Carbonic anhydrase and Phosphodiestrase-1 activity. Oxyberberine was initiate pointedly energetic (with IC50 values of 140.2±1.21) against urease enzyme inhibition assay while low activity against carbonic anhydrase, phosphodiestrase-I, and α-chymotrypsin. Urease showed active binding and next molecular modelling was performed. It was observed that the Oxyberberine shows significant integration with the active site of the jack bean urease. Different softwares were used like Autodck Vina and i-GEMDOCK v 2.1 for the docking analysis. The method of docking was accustomed with co-crystallized ligand of receptor P-gp. Different softwares such as PyMOL, discovery studio visualizer software and LIGPLOT+ version v.1.4.5 Autodock Vina and i GEMDOCK version 2.1 was used for the docking analysis.