PROSPECTIVE UTILIZATION OF ANTHERAEA MYLITTA COCOONASE AND ITS MOLECULAR HARMONY WITH NATURE.

fibroin. It is observed that during pupa-adult metamorphosis moth secretes proteolytic enzyme cocoonase. This proteolytic enzyme is one of the un-utilized by-products of tasar silk industry. Cocoonase is having efficacy to soften anterior portion of cocoons and adult moths comes out from silk cocoons. Therefore, a study towards utilization of cocoonase in cocoon cooking has been conducted. It is reported that, several sericigenous insects including A. mylitta exude a proteolytic enzyme cocoonase as they near the final stages of their metamorphosis. This proteolytic enzyme makes anterior portion of cocoon soft which facilitates the moth exit from cocoon (Kafatos and 1964; Felsted et al., 1973; Pandey et al., 2011; Unajak et al ., 2015). Studies on cocoonase of different sericigenous insects have been made by number of researchers (Felsted et al., 1973, and Li 2002; Wang et al ., 2005; Wang at al., 2005a; Yu-Dan, 2008; Wu et al ., 2008; Wang, et ., Rodbumrer et al ., 2012; Fukumori al ., 2014; Geng et al ., 2014; Unajak et al ., 2015) without much correlation with its possible-efficacy in cocoon cooking. It is proven that cocoonase directly acts on the sericin protein without affecting the fibroin protein. The SDS-PAGE analysis of freshly collected cocoonase showed molecular weight around 25-26 kDa. Although, our initial result indicates cocoon processing/cooking in cocoonase retains the tasar silk yarn natural beautiful unique tasar silk color, softness and luster (Pandey et al., 2011) but utilization of A. mylitta cocoonase and its molecular harmony with nature has not been studied much. Therefore, present study was conducted to study the prospective utilization of Antheraea mylitta cocoonase and its molecular harmony with nature. enzymatic SDS-PAGE of purified cocoonase showed molecular 25-26 kDa Per insect cocoonase collection volume is 500 to 800µl volume with 221µg/ml concentration. Recognition of sericin as natural substrate of cocoonase and it is established that cocoonase directly acts on the sericin without affecting the fibroin protein . Elemental-mapping/profiling of secreted cocoonase, suitable condition for specific activity of cocoonase, utilization of purified native cocoonase for cocoon softening has been done. Results indicate that cocoonase reutilization (one time) is also possible. SEM study showed marked variation in silk fiber surface when cocoon softening was done using cocoonase. Maldi Tof-Tof (MS and MSMS) data A& B of A. mylitta cocoonase MS 1320.477 showed similarities with cocoonase/proteolytic enzyme of other sericigenous insects. Molecular matching of Antheraea spp. cocoonase was also performed in order to identify the possible varients available in nature. Silk cocoons mainly consist of two main proteins, sericin and fibroin, with a glue-like layer of sericin coating two singular filaments of fibroin. Cocoon shell contains around 68-74% fibroin and 21-26% sericin. It is noticed that, cocoonase directly acts on the sericin protein without affecting the fibroin protein (silk fiber). It obviously indicates that, sericin is excellent natural-substrate for cocoonase. Based on sequential change in secreted cocoonase concentration specific time for cocoonase expression has been identified. Interestingly, the chronological changes in native cocoonase concentration were observed. Maximum concentration was observed in early collection followed by mid and late. The cocoonase secretion was more during first 30 minutes. Around 260-300µl of cocoonase was secreted during this period, 150-200µl cocoonase was secreted