Structural insight on the liquid silk from the middle silk gland of non-mulberry silkworm Antheraea assamensis.

This study highlights the preliminary characterization of liquid silk from the middle silk gland (MSG) along with the in-silico analysis of the sericin protein of a less explored non mulberry silkworm Antheraea assamensis which is endemic to the North Eastern region of India. Various biophysical methods have been applied to elucidate the conformational patterns of the liquid silk present inside the MSG without removing the sericin layer. This will help us to know the actual features of the in vivo transitional status of the silk in the MSG which travel towards the anterior silk gland (ASG) prior to spinning. The SDS PAGE analysis represented the existence of the both fibroin and sericin bands in the sample. The structural pattern of the MSG liquid silk as revealed by various methods denoted the occurrence of ß-sheet component along with some random coil and ß-turn components which in turn suggests the transitional state of the liquid silk attributed to the existence of both the crystalline and amorphous contents. The thermo gravimetric study and the aggregation behavior analysis results proposed the occurrence of intermolecular hydrogen bonding between the sericin and fibroin in the MSG. This study will sensitize the better understanding of the behavior of the liquid silk in the MSG of non-mulberry silkworm A. assamensis and will open avenues for various application-based studies of this silk.Communicated by Ramaswamy H. Sarma.

Composition and in silico structural analysis of fibroin from liquid silk of non-mulberry silkworm Antheraea assamensis.

Silk is spun from the liquid precursor known as liquid silk secreted from the posterior part and stored in the silk gland lumen with occurrence of many momentary events. The liquid silk in the silk gland is transformed to the spun silk fibre. In this study the elucidation of the protein components of liquid silk from the posterior part of the silk gland (PSG) of saturniid silkworm Antheraea assamensis along with its structural characterization has been reported. The 3D model of the N-terminal amorphous portion with some repeat crystalline motifs (19-255) of core protein fibroin has also been constructed. 1D and 2D electrophoresis revealed the homo-dimeric structure of the silk protein. Secondary structure analysis by Circular dichroism, FTIR spectroscopy showed α helical structural component as predominant conformation in the liquid silk. The crystalline structure investigated through X ray diffraction (XRD) analysis also revealed the presence of less ordered amorphous α helical conformation in the liquid silk. The 3D structural model proposed of the residues from 19 to 255 has revealed structural stability throughout the molecular dynamics simulation process. This study will provide the detailed structural information and in silico analysis of the core protein present in the liquid silk of PSG.

Variations in the Metallic Ion Concentration in the Silk Gland and Cocoon of Silkworm Antheraea assamensis helfer.

The natural spinning process in silkworms involves the conformation transition of the liquid silk protein present in the silk gland to make fiber. This conformation transition is influenced by different factors, and some studies suggested that changes in the metallic ion concentrations is one of them (Zhou et al. 2005). This study investigated the changes in the metallic ion compositions in the silk glands (before and during spinning) and cocoons of non-mulberry silkworm Antheraea assamensis helfer. Intact silk glands were dissected from mature 5th instar A. assamensis larvae. The glands were rinsed with deionized water and divided into five divisions: posterior silk gland (PSG), middle silk gland (MSG), anterior silk gland (ASG), posterior middle (PM) and anterior middle (AM) division of silk gland. Cocoon pieces and the gland parts were dried and digested in acid mixture to quantify the metallic contents in an atomic absorption spectrophotometer (Shimadzu, AA7000). We determined seven metals (Na, K, Mg, Ca, Cu, Zn, Fe) present in the different parts of the secretory pathway as well as in the fibers of A.assamensis. Our results suggested that the concentrations of Mg, Ca, Na, and K were more abundant in the gland than the Cu and Zn. Fe concentration was found comparatively less in amount in the gland. Amount of Ca found to be higher in the cocoons. The differences in the metallic ion concentrations in the gland parts before and during spinning suggested the possibility of their role in the formation of silk thread from luminal silk.