Changes in molecular weight and secondary structure of eri silk during alkali degumming and silk powdering were studied. An increase in silk degumming intensity, through increased alkali concentration, treatment temperature, and time, reduced the fibroin molecular weight and, therefore, the fiber tenacity, but at the same time, increased the beta-sheet fraction. These changes reduced the time required to mill the degummed silk fibers into fine powders. Mechanical forces used in wet attritor and air jet milling disturbed intermolecular packing along the direction of side chains, but the conformation remained essentially beta-sheet even in the sub-micron silk particles. Dry milling drastically reduced molecular weight and changed the conformation of the fibroin chains. The rate of the spontaneous conformation transition in regenerated fibroin solution prepared from fibers and powders increased with a decrease in fibroin molecular weight, affecting the time fibroin solutions could be stored before gelling. Overall, the study showed that molecular weight and secondary structure of silk powders could be manipulated by suitably changing the degumming and milling conditions. It also suggests that wet media milling and air jet milling are better than dry media milling to prepare less degraded and more crystalline ultrafine silk particles. (C) 2010 Wiley Periodicals, Inc. J Appl Polym Sci 119: 1339-1347, 2011