复合酶法高效制备紫菜原生质体的研究  

Research of Efficient Preparation of Laver Protoplasts by Compound Enzymatic Method

作　　者：穆惠敏 朱常亮[2] 沈照鹏[2,3] 孟蕾 江晓路 MU Hui-min1, ZHU Chang-liang2, SHEN Zhao-peng2,3 ,MENG Lei4 , JIANG Xiao-lu1,3,4(1.College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China; 2.College of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; 3.Qingdao Marine Biomedical Research Institute, Qingdao 266071, China; 4.State Key Laboratory of Bioactive Seaweed Substances,Qingdao Brightmoon Seaweed Group Co.Ltd., Qingdao 266400, China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]中国海洋大学医药学院,山东青岛266003 [3]青岛海洋生物医药研究院,山东青岛266071 [4]青岛明月海藻集团有限公司海藻活性物质国家重点实验室,山东青岛266400

出　　处：《食品工业科技》2018年第19期142-146,共5页Science and Technology of Food Industry

基　　金："十三五"海洋经济创新发展示范城市项目(2016QD003);青岛市海洋医药科技创新中心项目(2017-CXZX01-3-13)

摘　　要：以紫菜(Porphyra yezoensis)为研究对象,通过复合酶水解获得原生质体,研究酶解温度、加酶量、酶解时间、摇床转速、盐度、复合酶中组分比例、金属离子对酶解后原生质体数的影响。在单因素试验的基础上,以原生质体数为指标,选取因子与水平进行正交实验,对酶解条件进行优化研究。结果表明,最佳酶解条件为:酶解温度28℃、转速125 r/min、纤维素粗酶含量3%、琼胶酶12 m L、盐度4%、酶解时间100 min。同时发现一些金属离子对原生质体制备有抑制作用。在最优酶解条件下,得到的原生质体活率较高,即1 g紫菜样品经酶解得到的原生质体活率高达(16.88±0.22)×10~6个/g。综上,复合酶法制备紫菜原生质体反应条件温和、成本低廉,为紫菜的遗传育种以及在水产养殖中的应用研究提供理论依据。Laver（ Porphyra yezoensis） was chosen for enzymatic hydrolysis,and protoplasts were obtained by enzyme hydrolysis.Effects on the enzyme of temperature,the amount of enzyme,reaction time,rotary speed,salinity,the proportion of components in the enzyme and metal ion were studied by the number of laver protoplasts.Factors and levels were selected on the basis of single factor test,the optimization of hydrolysis conditions were obtained by orthogonal experiment with the number of protoplasts as the index. The results indicated that the optimal hydrolytic conditions were： Temperature 28 ℃,rotary speed125 r/min,content of cellulose crude enzyme 3%,the agar enzyme 12 mL,salinity 4% and hydrolysis time 100 min.Protoplast formation inhibited by some metal ions were found simultaneously.Protoplasts obtained under the optimal hydrolysis conditions were high live rate,the number of protoplasts obtained from 1-g porphyra sample was（ 16.88 ± 0.22） × 10^6 cells/g.

关 键 词：海藻解壁酶 紫菜 复合酶法 原生质体 

分 类 号：TS201.1[轻工技术与工程—食品科学]