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Personals offer an essay and the ability to add up to ten photos. The cocoons containing relatively higher amounts of protein-tannin complexes in the form of proanthocyanidins Pandey and Goel, ; Pandey, , have to be softened by more drastic boiling off techniques Pandey and Goel, Generally the cocoons are cooked in presence of strong alkali agent or other harsh chemicals Tikoo and Goel, ; Das, ; Iizuka et al. Since the chemical methods reduce the quality of the tasar silk thread in many ways Tikoo and Goel, , an alternative method for the oak tasar cocoon cooking based on the proteolytically active pineapple extract may be developed for better results.
The pineapple extract may be used in cooking of the silk cocoon to soften it by decomposing or partially solubilizing the proteinaceous silk gum sericin which is involved in binding the fibroin silk strands together in the cocoon shell. In the present investigation, a study was undertaken to enzymologically characterize the pineapple extract having proteinase activity with the aim of developing an effective oak tasar Antheraea proylei J.
Materials and Methods Pineapple pulp Fresh and ripe mature and yellow fruit of pineapple Ananas comosus L. Queen was purchased from the markets in and around Imphal, Manipur, India. The pineapple fruit pulp was prepared from the fruit by first detaching the crown and stem parts, and then slicing off the skin part.
Silk cocoon The cocoons produced by the oak tasar silkworm Antheraea proylei J. Three specimen cocoons are shown in Figure 1. Preparation of pineapple extract The pulp 15 g of ripe mature and yellow pineapple fruit was homogenized with distilled water mL. Proteinase assay The proteinase activity of the pineapple extract was assayed by a modification of the azocasein method of Rowan and Buttle, in which the time-dependent release of azo-dye-coupled-TCA-soluble-peptide fragments from the proteinase substrate azocasein was monitored.
The assay mixture was constituted by mixing 1. The assay mixture was found to have a fixed pH value ranging from 3. The reaction was started with the addition of the substrate.
After 30 min interval, the reaction was arrested by adding 0. The protein precipitation was allowed to complete by cooling the resulting reaction mixture in ice for 5 min and then centrifuged to collect the supernatant. A corresponding blank was run by adding TCA to the pineapple extract prior to the mixing with the azocasein solution. Determination of effect of sodium carbonate The proteinase activity of the pineapple extract was determined as a function of sodium carbonate concentration.
The proteinase assay mixture was constituted by mixing 1. The rest of the procedure was same as in Proteinase assay. Corresponding blank at each concentration of sodium carbonate was run by adding TCA to the pineapple extract prior to mixing with azocasein. Determination of optimum temperature The proteinase activity of the pineapple extract was determined as a function of temperature.
The assay mixture, constituted by mixing 1. After 30 min incubation, the reaction was arrested by addition of 0. Corresponding blank at each temperature was run by adding TCA to the pineapple extract prior to mixing with azocasein. Similar experiment was repeated with reaction mixture containing 9. Determination of stability characteristics Thermal stability. Corresponding blank at each temperature was run by adding TCA prior to addition of azocasein to the pineapple extract.
Similar experiment was repeated by incubating the pineapple extract containing 9. Time course of inactivation.
At different time intervals, 1. Corresponding blank at each time interval was run by adding TCA prior to addition of azocasein to the pineapple extract. Similar experiment was repeated by taking the pineapple extract containing 9.
Determination of effect of cocoon extract Effect on proteinase activity.