Miyamoto I, Nagase S
J Biochem. 1984 May;95(5):1331-6. doi: 10.1093/oxfordjournals.jbchem.a134739.
Hyaluronic acid was purified from the horny layer of guinea pigs and its biochemical and physical properties were studied. The horny layer, obtained by applying n-hexadecane to guinea pig skin, was digested with pronase, and glycosaminoglycans in the digest were separated from UV-absorbing material by Sephadex G-75 chromatography (sample A, 17.5 mg). On DEAE-Sephadex chromatography, the fraction obtained with 0.5 M NaCl was found to contain 94% of the total uronic acid. This fraction, consisting mainly of hyaluronic acid, was dialyzed and lyophilized (sample B, 12.5 mg). Sample B, consisting of 26.1% uronic acid and 27.0% glucosamine on a dry weight basis, could be digested completely with Streptomyces hyaluronidase. Sample B had a low reduced viscosity which showed almost no concentration dependence. The intrinsic viscosity of sample B was 0.83 dl/g and its molecular weight, calculated from its viscosity, was 34,000. Sample B was eluted from Sepharose CL-6B as a broad peak between the void volume and the total column volume. The enzyme levels of hyaluronidase, beta-glucuronidase, and beta-N-acetylglucosaminidase in the n-hexadecane treated guinea pig skin increased to 1.7 to 2.5 fold those of controls after 6 days of the experiment. These results suggested that hyaluronic acid in the horny layer of n-hexadecane treated guinea pig skin might be degraded by hyaluronic acid degrading enzymes in the hyperkeratinized tissue.
从豚鼠角质层中纯化出透明质酸，并对其生化和物理性质进行了研究。通过将正十六烷涂覆在豚鼠皮肤上获得角质层，用链霉蛋白酶进行消化，消化液中的糖胺聚糖通过Sephadex G - 75色谱法与紫外线吸收物质分离（样品A，17.5毫克）。在DEAE - Sephadex色谱上，用0.5M NaCl洗脱得到的级分中发现含有总糖醛酸的94%。该级分主要由透明质酸组成，经过透析和冻干（样品B，12.5毫克）。以干重计，样品B含有26.1%的糖醛酸和27.0%的葡萄糖胺，可被透明质酸链霉菌酶完全消化。样品B的比浓粘度较低，几乎不显示浓度依赖性。样品B的特性粘度为0.83 dl/g，根据其粘度计算出的分子量为34,000。样品B从Sepharose CL - 6B上以一个宽峰的形式洗脱，介于空体积和总柱体积之间。在实验6天后，经正十六烷处理的豚鼠皮肤中透明质酸酶、β - 葡萄糖醛酸酶和β - N - 乙酰氨基葡萄糖苷酶的酶水平增加到对照的1.7至2.5倍。这些结果表明，经正十六烷处理的豚鼠皮肤角质层中的透明质酸可能被过度角化组织中的透明质酸降解酶降解。
