Second‐generation biofuels prove to be a distinctive and renewable source of sustainable energy and cleaner environment. The current study focuses on the exploration and identification of four nonedible sources, that is, Brassica oleracea L., Carthamus oxyacantha M.Bieb., Carthamus tinctorius L., and Beaumontia grandiflora Wall., utilizing light microscopy (LM) and scanning electron microscopy (SEM) for studying the detailed micromorphological features of these seeds. LM revealed that size ranges from 3 to 20 mm. furthermore, a great variety of color is observed from pitch black to greenish gray and yellowish white to off white. Seeds ultrastructure study with the help of SEM revealed a great variety in shape, size, color, sculpturing and periclinal wall shape, and so on. Followed by the production of fatty acid methyl esters from a novel source, that is, seeds oil of Brassica oleracea L. (seed oil content 42.20%, FFA content 0.329 mg KOH/g) using triple metal impregnated montmorillonite clay catalyst (Cu‐Mg‐Zn‐Mmt). Catalyst was characterized using SEM–EDX, FT‐IR. Maximum yield of Brassica oleracea L. biodiesel (87%) was obtained at the conditions; 1:9 of oil to methanol ratio, 0.5 g of catalyst, 5 hr reaction time, and 90°C of temperature. Synthesized biodiesel was characterized by FT‐IR, GC–MS, and NMR. Fuel properties of the Brassica oleracea L. FAMES were determined and found in accordance with ASTM standards.