Momordica charantia is a plant of the Cucurbitaceae family which is known as bitter gourd and grows widely in tropical and subtropical regions of the world. The various properties of bitter gourd are closely related to the substances or compounds they contain. Bitter gourd contains many secondary metabolites including alkaloids, terpenoids, steroids and flavonoids. These chemical compounds are spread in several parts of the plant such as roots, stems, leaves, and fruit. Apart from containing several organic compounds, bitter melon also contains the phytohormone gibberellin which is quite potential and useful as a biostimulant. The aim of the research was to isolate, purify, and determine the content of gibberellin (GA-3) contained in bitter gourd (M. charantia), and to determine its effect on the vegetative and generative growth of soybean plants, as well as the effect of GA-3 application on the nutritional content of soybean seeds. resulting from. Apart from that, it was also observed the effect of GA-3 application on the chlorophyll content of soybean plants, as well as the activity of the alpha-amylase enzyme in treated soybean seeds. The process of isolation and purification of GA-3 was carried out by thin layer chromatography and adsorption column chromatography on Silica Gel G-60. Characterization of GA-3 was carried out by Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC), Fourier Transform Infrared (FT-IR), and Mass Spectroscopy (MS). RP-HPLC and FTIR results from gibberellin GA-3 isolates showed retention times and wave numbers that were relatively the same as GA-3 standards. From the results of the study it was found that the content of the gibberellin GA-3 hormone in bitter gourd was 4.12 mg/g dry weight. The GA-3 isolate had a significant effect on the growth of soybean plants in both the vegetative and generative phases for each parameter observed compared to the control and 1% NPK fertilizer. The protein content in soybean seeds was 38.17%. The chlorophyll content at the end of the vegetative and generative phases were 33.34 and 11.5 CCI, respectively. The addition of GA-3 isolate increased the activity of the α-amylase enzyme in soybean seeds compared to the control with an average of 271.65 U/mL.