Androstenedione is the main product after phytosterol fermentation by Mycobacteria. Bisnorcholenaldehyde (BA; 3-Oxopregn-4-ene-20-carboxaldehyde) is one of the by-products produced during fermentation. Bisnorcholenaldehyde is an essential intermediate in the production of progesterone, which is used to treat threatened abortion or habitual abortion disease caused by luteal insufficiency. It is a precursor of androgen, estrogen, glucocorticoid and intermediates of steroid hormone synthesis[1]. In addition, BA is used in magnetic-based graphene composites with steroidal diamine dimer as a potential drug in hyperthermia cancer therapy.