Optimizing the sulfation-modification system for scale preparation of chondroitin sulfate A
Optimizing the sulfation-modification system for scale preparation of chondroitin sulfate A
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规模化制备硫酸软骨素A的硫酸化改性体系优化
DOI：
10.1016/j.carbpol.2020.116570
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发表时间：
2020-10-15
影响因子：
11.2
通讯作者：
Kang, Zhen
中科院分区：
文献类型：
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作者：
Jin, Xueron;Li, Qing;Kang, Zhen
从动物组织中提取的硫酸软骨素(CS)已被广泛用于治疗骨关节炎的营养和医药产品。在这里，我们建立了一种高效的硫化修饰体系，用于体外大规模制备环孢素A。首先，随着伴侣相扑的融合，C4ST的表达水平提高了30倍。在辅因子PAPs的再生过程中，甘油作为蛋白质稳定剂可以提高大鼠AST IV的稳定性。然后利用多肽连接物或蛋白质支架将AST IV和C4ST组装成人工复合体，使酶和PAPs在空间上更接近，从而提高了软骨素硫化的催化效率。最终将该体系扩大到1个L体系，并在24 h内将15g软骨素转化为CsA，转化率为98%。本研究进一步向工业化生产不同硫酸盐化程度的CSA迈进了一步。
Chondroitin sulfate (CS) extracted from animal tissues has been widely used as nutraceutical and pharmaceutical products for osteoarthritis treatment. Here we developed an efficient sulfation-modification system for large scale preparation of CSA in vitro. First, the expression level of C4ST was improved by 30 times with fusion of the chaperone SUMO. Then, glycerol as a protein stabilizer was found to improve rat AST IV stability during the regeneration of cofactor PAPS. Then peptide linkers or protein scaffolds were employed to assemble AST IV and C4ST into artificial complexes to bring the enzymes and PAPS spatially closer and enhance the catalytic efficiency of chondroitin sulfation. Eventually, the system was scaled up to 1 L system and 15 g chondroitin was converted to CSA in 24 h, with a 98 % conversion. The present study made a step further towards the industrial production of CSA with different sulfation degrees.