摘要
为获得优质的黑木耳胶原蛋白,本研究通过单因素实验和响应面法对碱性蛋白酶法提取黑木耳胶原蛋白的工艺条件进行优化,并采用盐析、DEAE-52离子交换层析、Sephadex G-75凝胶过滤等方法对黑木耳胶原蛋白进行分离纯化。结果表明:在酶解温度55℃、pH 8.5条件下优化得到的黑木耳胶原蛋白最佳提取条件为:碱性蛋白酶(2240 IU/mg)添加量7%、料液比1∶68 g/mL、酶解时间2.0 h,此条件下,黑木耳胶原蛋白得率为0.65%;最佳盐析条件为:4 mol/L NaCl、24 h、15℃;最佳DEAE-52分离条件为:洗脱时间160 min、上样浓度300μg/mL、上样pH 7.2、NaCl洗脱浓度0.2 mol/L;经Sephadex G-75凝胶过滤层析后胶原蛋白纯度可达到72.6%,胶原蛋白回收率74.3%。因此,碱性蛋白酶法及盐析、DEAE-52、Sephadex G-75层析可有效提取纯化黑木耳胶原蛋白,为黑木耳胶原蛋白的深入研究及应用开发提供了理论依据。
In order to obtain high quality collagen from black fungus,the extraction conditions of collagen by alkaline protease were optimized by single factor experiment and response surface method.The black fungus collagen was separated and purified by salting out,DEAE-52 ion exchange chromatography and Sephadex G-75 gel filtration.The results showed that the optimal extraction conditions were as follows:alkaline protease(2240 IU/mg)7%,solidliquid ratio 1∶68 g/mL,and enzymatic hydrolysis time 2.0 h.Under these conditions,the yield of collagen was 0.65%.The optimal salting-out conditions were as follows:4 mol/L NaCl for 24 h at 15℃.The optimal DEAE-52 separation conditions were as follows:elution time 160 min,loading concentration 300μg/mL,loading pH 7.2,and NaCl elution concentration 0.2 mol/L.Being purified with Sephadex G-75 gel filtration chromatography,the purity of collagen was 72.6% and the recovery of collagen was 74.3%.Therefore,the alkaline protease method and salting out,DEAE-52 and Sephadex G-75 chromatography could effectively extract and purify the collagen from black fungus.The study provides theoretical basis for the further study and application of the collagen from black fungus.
作者
毕海鑫
林雨泽
孟桥
那治国
BI Haixin;LIN Yuze;MENG Qiao;NA Zhiguo(College of Food Engineering,East University of Heilongjiang,Harbin 150066;College of Food Engineering,Harbin University of Commerce,Harbin 150028)
出处
《中国食品添加剂》
CAS
北大核心
2023年第9期56-65,共10页
China Food Additives
基金
黑龙江省自然科学基金联合引导项目(LH2020C078)
黑龙江省重点研发计划指导类项目(GZ20210166)
哈尔滨市科技计划自筹经费项目(ZC2022ZJ020002)
黑龙江东方学院科研项目(HDFKY200101)。
关键词
黑木耳
胶原蛋白
碱性蛋白酶
分离纯化
black fungus
collagen protein
alkaline protease
separation and purification
