Production of Serine Protease Inhibitors from Momordica Dioica Cell Cultures

Abstract

Protein molecules which bind and inhibit the activity of serine protease enzymes are known as serine protease inhibitors. These are proteolytic enzymes which possess triad of aspartic acid, histidine and serine in their active site. They have been found in microbes, plant tissues, animal tissues and fluids and insects. In plants, they play a vital role in defence mechanism against predators and pathogens. Serine protease inhibitors are mostly detected in seed extracts. The present study aims the development of callus cultures of Momordica dioica for the production of serine protease inhibitors which may have potential as bioinsecticide. Callus cultures of M. dioica were established from various explants which obtained from in-vitro as well as in- vivo growing plants. M. dioica seedlings were developed under in-vitro condition through germination of stored seeds on water agar medium. Sixty percent germination was achieved. Leaves obtained from outdoor grown plants were proved to be the best explants for callus formation with 60% rate of callus induction. Protein profile of callus and media samples was analyzed through SDS-PAGE and two proteins were found to be similar and expressed in seeds also along with several others. Trypsin inhibitory activity was reported in callus extract (91%) and media samples (54%). Since trypsin inhibitory activity was also observed in the protein extracted from the medium, it is believed that the protein involved is extracellular.