A novel and environmentally friendly method for preserving and depilating sheepskin : comprehensive physical, biochemical and molecular analyses : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistry at Massey University, Manawatu, New Zealand

Abstract

The first step of leather processing, depilation – in other words, removing hair from skins – accounts for one-third of the leather-making industrial waste due to the production of sulfide and alkaline water waste from the process. This study describes a method that preserves and depilates unwashed sheepskins using milk or milk by-products, including whey and permeate. It doesn’t require the use of harsh chemicals or temperature control, and results in skin that is ready to be tanned. In order to evaluate the products of this process, the depilated skin surface was evaluated not only by eye, but with scanning and transmission electron microscopy (SEM and TEM) which showed there was no apparent damage to the grain or fraying of the collagen bundles. The depilated skin was also processed to leather which was subjected to tear, tensile and shrinkage measurements which were shown to be almost identical to leathers made using the traditional process. Quantitative biochemical analyses, including collagen quantitation, collagen crosslink analysis and glycosaminoglycan (GAG) analysis of sheepskins depilated with this process showed no significant differences in both collagen, and collagen crosslink concentrations in contrast to the 10-fold increase seen in the GAG concentration. A quantitative proteomic analysis showed there was a higher retention of proteins found in the basement membrane of the skin, supporting the observation that permeate depilated skins were smoother than their sulfide depilated counterparts and produced leather with a superior surface. It also showed the loss of specific proteins around the hair follicle, hinting at a possible mechanism for depilation. To further investigate this a microbiological survey of the process was conducted. Traditional culturing methods were used to isolate and identify microorganisms present in the depilation solution after the wool had been removed. Two bacterial species (Lactococcus lactis and Lactobacillus plantarum) and two fungal species (Geotrichum candidum and Yarrowia lipolytica), were routinely identified, all of which are known to secrete a number of hydrolytic enzymes and antimicrobial compounds. This was followed by a metagenomic study of changes in the microbial community over the time course of the depilation. Although there were only 13 dominant bacterial genera identified during this study, the biggest change was a concomitant increase in the relative abundance of Lactococcus lactis that matched the decrease in Acinetobacter sp. by the end of the depilation treatment, controlling the proliferation of other putrefying organisms. In conclusion, this preliminary study has paved the way for the development of a depilation process that is kind to the environment, but more research is needed to investigate its potential use with other animal skins.