Kim Chong-Tai, Gujral Naiyana, Ganguly Advaita, Suh Joo-Won, Sunwoo Hoon H
Food Bio-Nano Research Group, Korea Food Research Institute, 516, Baekhyun-Dong, Bundang-Ku, Seongnam-Si, Gyeonggi-Do 463-746, Republic of Korea.
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11361 - 87 Avenue, Edmonton, Alberta T6G 2E1, Canada.
Biotechnol Rep (Amst). 2014 Jul 19;4:14-20. doi: 10.1016/j.btre.2014.07.004. eCollection 2014 Dec.
Chondroitin sulphate (CS), a major glycosaminoglycan, is an essential component of the extracellular matrix in cartilaginous tissues. Wapiti velvet antlers are a rich source of these molecules. The purpose of the present study was to develop an effective isolation procedure of CS from fresh velvet antlers using a combination of high hydrostatic pressure (100 MPa) and enzymatic hydrolysis (papain). High CS extractability (95.1 ± 2.5%) of total uronic acid was obtained following incubation (4 h at 50 °C) with papain at pH 6.0 in 100 MPa compared to low extractability (19 ± 1.1%) in ambient pressure (0.1 MPa). Antler CS fractions were isolated by Sephacryl S-300 chromatography and identified by western blot using an anti-CS monoclonal antibody. The antler CS fraction did not aggregate with hyaluronic acid in CL-2B chromatography and possessed DPPH radical scavenging activity at 78.3 ± 1.5%. The results indicated that high hydrostatic pressure and enzymatic hydrolysis procedure may be a useful tool for the isolation of CS from antler cartilaginous tissues.
硫酸软骨素(CS)是一种主要的糖胺聚糖,是软骨组织细胞外基质的重要组成部分。马鹿鹿茸是这些分子的丰富来源。本研究的目的是开发一种使用高静水压(100兆帕)和酶解(木瓜蛋白酶)相结合的方法,从新鲜鹿茸中有效分离CS。与常压(0.1兆帕)下的低提取率(19±1.1%)相比,在100兆帕、pH 6.0条件下用木瓜蛋白酶孵育(50℃下4小时)后,总糖醛酸的CS提取率较高(95.1±2.5%)。鹿茸CS级分通过Sephacryl S-300色谱法分离,并使用抗CS单克隆抗体通过蛋白质印迹法进行鉴定。鹿茸CS级分在CL-2B色谱中不与透明质酸聚集,并且具有78.3±1.5%的DPPH自由基清除活性。结果表明,高静水压和酶解方法可能是从鹿茸软骨组织中分离CS的有用工具。