碱溶酸沉盐析法纯化水不溶烟叶蛋白质  

Purification of water-insoluble tobacco proteins by alkaline dissolution,acid precipitation and salting-out method

作　　者：张建平 蓝洪桥 黄梦贤 刘秀彩 黄华发 陈伟珠 郑泉兴 方华 刘泽春 ZHANG Jianping;LAN Hongqiao;HUANG Mengxian;LIU Xiucai;HUANG Huafa;CHEN Weizhu;ZHENG Quanxing;FANG Hua;LIU Zechun(Technology Center,China Tobacco Fujian Industrial Co.,Ltd.,Xiamen 361021,Fujian,China;Technical Innovation Center for Utilization of Marine Biological Resources,Third Institute of Oceanography,Ministry of Natural Resources,Xiamen 361005,Fujian,China;College of Biology and Environment,Zhejiang Wanli University,Ningbo 315100,Zhejiang,China;Xiamen Ocean Vocational College,Xiamen 361102,Fujian,China)

机构地区：[1]福建中烟工业有限责任公司技术中心,福建省厦门市361021 [2]自然资源部第三海洋研究所海洋生物资源开发利用工程技术创新中心,福建省厦门市361005 [3]浙江万里学院生物与环境学院,浙江省宁波市315100 [4]厦门海洋职业技术学院,福建省厦门市361102

出　　处：《烟草科技》2024年第11期18-25,共8页Tobacco Science & Technology

基　　金：福建中烟工业有限责任公司重点项目“纤维素酶在烟草中作用效果表征技术的研究及应用”(FJZYKJJH2023ZD005);厦门海洋与渔业局专项资金项目“海带中三种功能组分原料研发及产业化示范”(23CZP009HJ11)。

摘　　要：为了匹配烤烟蛋白质检测对照品的同源度,制备了烤烟来源的高质量分数蛋白质,进行了烤烟水不溶烟叶蛋白质的提取及纯化工艺研究,并中试规模制备了水不溶烟叶蛋白质粗提物。分别考察了pH值、亚氯酸钠的质量分数、加热温度和硫酸铵加入量4个单因素对水不溶烟叶蛋白质粗提物纯化效果的影响,采用凯氏定氮法检测样品中蛋白质的质量分数,采用色差仪检测并计算出明度值(L*),红度值(a*),黄度值(b*)和总色差值(ΔE*_(ab))。结果表明:①最佳纯化工艺为将5 g水不溶烟叶蛋白质粗提物,采用50 mL pH为8的3%(质量分数)亚氯酸钠反应液,在60℃条件下反应30 min,再加入10 g硫酸铵盐析,得到纯化过的水不溶烟叶蛋白质样品。②在最佳纯化工艺条件下,所制备得到的水不溶烟叶蛋白质样品的质量分数为88.41%,收率为12.57%,且与对照组的ΔE*_(ab)值最大。To match the homology of protein detection reference materials from flue-cured tobacco,proteins with higher mass fractions were prepared from flue-cured tobacco sources,and the extraction and purification processes of water-insoluble tobacco proteins were studied.In addition,a crude extract of water-insoluble proteins from tobacco leaves was prepared on a pilot scale.The effects of four single factors,including pH,mass fraction of sodium chlorite,heating temperature,and addition amount of ammonium sulfate,on the purification effect of crude water-insoluble tobacco proteins were investigated.The mass fractions of water-insoluble tobacco proteins were determined by the Kjeldahl method,and the values of L*,a*,b*,andΔE*_(ab)were measured and calculated using a colorimeter.The results showed that:1)The optimal purification process was achieved by using 5 grams of crude water-insoluble tobacco proteins,which were reacted with 50 mL of 3%(mass fraction)sodium chlorite solution at pH 8 and 60℃for 30 minutes,and then salted out with 10 grams of ammonium sulfate.2)Under the optimized processing condition,the mass fraction and yield of water-insoluble tobacco proteins were 88.41%and 12.57%,respectively,and theΔE*_(ab)value between the sample and the control group was the largest.

关 键 词：烤烟 水不溶烟叶蛋白质 总色差值(ΔE*_(ab)) 单因素实验 盐析法 

分 类 号：TS411.1[农业科学—烟草工业]