杨会成,胡世伟,周小敏,等.金枪鱼骨胶原蛋白肽螯合钙制备工艺及对MG-63细胞碱性磷酸酶分泌的影响[J].中国海洋药物,2020,39(1):09-18.
金枪鱼骨胶原蛋白肽螯合钙制备工艺及对MG-63细胞碱性磷酸酶分泌的影响
Preparation of collagen peptide from tuna bone chelated calcium and its effects on alkaline phosphatase secretion in MG-63 cells
投稿时间：2019-06-27  修订日期：2019-08-11
DOI：
中文关键词:  金枪鱼骨  胶原蛋白肽螯合钙  制备工艺  碱性磷酸酶
English Keywords:tuna bone  collagen peptide chelated calcium  preparation technology  alkaline phosphatase
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作者单位E-mail
杨会成 浙江海洋大学  
胡世伟* 浙江海洋学院 hushiweihai@163.com 
周小敏 浙江兴业集团有限公司  
李澄 浙江兴业集团有限公司  
郑万源 浙江兴业集团有限公司  
李世杰 浙江海洋大学  
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中文摘要:
      目的 为获得一种能够增加人成骨细胞碱性磷酸酶生成量的金枪鱼骨胶原蛋白肽螯合钙(collagen peptide from tuna bone chelated calcium, TCP-Ca)的工艺方法。方法 本文利用正交实验优化蛋白酶酶解法制备金枪鱼骨胶原蛋白肽的工艺,利用正交实验优化金枪鱼骨胶原蛋白肽和柠檬酸钙进行螯合反应的工艺,然后利用傅里叶变换红外光谱对TCP-Ca进行定性分析,同时检测TCP-Ca对MG-63细胞碱性磷酸酶生成量的影响。结果 金枪鱼骨胶原蛋白肽制备的最佳工艺为pH 9.0、酶解时间180 min、加酶量5000 U/g、酶解温度45 ℃；TCP-Ca制备的最佳工艺为温度35 ℃、pH 7.5、肽钙质量比6:1、时间40 min；金枪鱼骨胶原蛋白肽的氨基和羟基均参预了钙螯合反应；TCP-Ca比金枪鱼骨胶原蛋白肽更有利于增加MG-63细胞分泌碱性磷酸酶。结论 本文优化了TCP-Ca的制备工艺,发现其具有增加成骨细胞分泌碱性磷酸酶的生物活性。
English Summary:
      Objective To gain a technology for preparation of collagen peptide from tuna bone chelated calcium with the bioactivity of increase in alkaline phosphatase secreted by human osteoblast. Methods The technology for preaparation of collagen peptide from tuna bone (TCP) using protease digestion was optimized by orthogonal experiment. The technology for preaparation of TCP chelated calcium from TCP and calcium citrate was optimized by orthogonal experiment. TCP chelated calcium was analyzed using fourierStransformSinfraredSspectroscopy and its effect on alkaline phosphatase production from MG-63 cells was also investigated. Results The best preparation technology for TCP was: pH 9.0, enzymatic hydrolysis time 180 min, enzyme amount 5000 U/g, enzymatic hydrolysis temperature 45 ℃. The best preparation technology for TCP chelated calcium was: temperature 35 ℃, pH 7.5, ratio of peptide and caluium 6:1, time 40 min. Amino and hydroxyl of TCP were both chelated with calcium. TCP chelated calcium can increase more alkaline phosphatase secretion than TCP. Conclusion A kind of TCP chelated calcium with the bioactivity of increase in alkaline phosphatase secreted by human osteoblast was achieved, and the preparation technology was also optimized.
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