Institute of Biological Chemistry, Academia Sinica, No. 128, Academia Road, Section 2, Nankang, Taipei, 11529, Taiwan.
Graduate Institute of Biomedical Sciences, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan.
Sci Rep. 2022 Aug 29;12(1):14684. doi: 10.1038/s41598-022-18474-4.
Eggshell membrane (ESM), a plentiful biological waste, consists of collagen-like proteins and glycosaminoglycans (GAGs) such as hyaluronic acid (HA). Here we used a keratinase (oeMtaker)-mediated system to decompose ESM. The best reaction condition was established by incubating the solution containing oeMtaker, sodium sulfite, and ESM with a weight ratio of 1:120:600. ESM enzymatic hydrolysate (ESM-EH) showed a high proportion of essential amino acids and type X collagen peptides with 963-2259 Da molecular weights. The amounts of GAGs and sulfated GAGs in ESM-EH were quantified as 6.4% and 0.7%, respectively. The precipitated polysaccharides with an average molecular weight of 1300-1700 kDa showed an immunomodulatory activity by stimulating pro-inflammatory cytokines (IL-6 and TNF-α) production. In addition, a microorganism-based system was established to hydrolyze ESM by Meiothermus taiwanensis WR-220. The amounts of GAGs and sulfated GAGs in the system were quantified as 0.9% and 0.1%, respectively. Based on our pre-pilot tests, the system shows great promise in developing into a low-cost and high-performance process. These results indicate that the keratinase-mediated system could hydrolyze ESM more efficiently and produce more bioactive substances than ever for therapeutical applications and dietary supplements.
蛋壳膜（ESM）是一种丰富的生物废料，由类似于胶原蛋白的蛋白质和糖胺聚糖（GAG）组成，如透明质酸（HA）。在这里，我们使用角蛋白酶（oeMtaker）介导的系统来分解 ESM。通过在含有 oeMtaker、亚硫酸钠和 ESM 的溶液中以 1:120:600 的重量比进行孵育，确定了最佳反应条件。ESM 酶解产物（ESM-EH）表现出高比例的必需氨基酸和类型 X 胶原蛋白肽，分子量为 963-2259 Da。ESM-EH 中的 GAG 和硫酸化 GAG 含量分别定量为 6.4%和 0.7%。沉淀的多糖分子量平均为 1300-1700 kDa，具有刺激促炎细胞因子（IL-6 和 TNF-α）产生的免疫调节活性。此外，还建立了一种基于微生物的系统，通过 Meiothermus taiwanensis WR-220 水解 ESM。该系统中 GAG 和硫酸化 GAG 的含量分别定量为 0.9%和 0.1%。根据我们的初步试验，该系统在开发低成本、高性能工艺方面具有很大的应用前景。这些结果表明，角蛋白酶介导的系统可以更有效地水解 ESM，并产生更多的生物活性物质，用于治疗应用和膳食补充剂。
