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海带中抑藻活性化合物的分离及抑藻活性分析
引用本文:孙颖颖,朱文轩,周静,庄丽雯,毛奕淋,赵秀芳,王长海.海带中抑藻活性化合物的分离及抑藻活性分析[J].海洋学报,2022,44(2):143-154.
作者姓名:孙颖颖  朱文轩  周静  庄丽雯  毛奕淋  赵秀芳  王长海
作者单位:1.江苏海洋大学 江苏省海洋生物技术重点实验室,江苏 连云港 222005
基金项目:江苏省第五期“333工程”科研资助项目(BRA2020263);江苏省优势学科建设工程资助项目;江苏省研究生实践创新计划(SJCX20_1317);连云港市“海燕计划”;2018年江苏高校优秀中青年教师和校长境外研修计划。
摘    要:以2 000 g海带为原料、米氏凯伦藻为测试微藻,采用甲醇浸提、液液萃取、硅胶柱层析、Sephadax LH-20凝胶柱层析和硅胶薄层层析等分离方法,纯化可能具有抑藻活性的化合物,制备到5个薄层纯样品,B21(1.39 g)、B131(0.51 g)、B132(0.48 g)、C211(0.016 g)和C212(0.020 g)。经鉴定,B21、B131、B132、C211和C212依次为7-羟基庚酸、十四酸、十六酸、十八酸和油酸,均是第一次从海带中分离获得。此5种脂肪酸对塔玛亚历山大藻、强壮前沟藻、赤潮异弯藻、米氏凯伦藻、球形棕囊藻和中肋骨条藻的生长表现出选择性抑制作用,其中7-羟基庚酸具有较为广泛的抑藻活性。除塔玛亚力山大藻之外,在浓度为50 μg/mL时,7-羟基庚酸对其他5种赤潮微藻生长的抑制作用与硫酸铜和重铬酸钾对它们生长的抑制作用接近,生长抑制率超过71%(第4天);十四酸在浓度为50 μg/mL时对强壮前沟藻、米氏凯伦藻和中肋骨条藻生长也有明显的抑制作用,第4天时对它们的生长抑制率在57%以上。在此基础上,第一次确定了7-羟基庚酸、十四酸、十六酸和十八酸对强壮前沟藻、赤潮异弯藻、米氏凯伦藻、球形棕囊藻和中肋骨条藻生长的半抑制效应浓度(EC50~96 h),7-羟基庚酸对塔玛亚历山大藻生长的EC50~96 h也是第一次被确定。与硫酸铜和重铬酸钾对每种赤潮微藻生长的EC50~96 h比较,7-羟基庚酸和十四酸在抑制强壮前沟藻上具有明显的抑藻优势。

关 键 词:抑藻活性化合物    分离    海带    纯化    赤潮微藻
收稿时间:2021-07-31

Isolation,purification and analysis of antialgal activities of antialgal com-pounds from Laminaria japonica
Sun Yingying,Zhu Wenxuan,Zhou Jing,Zhuang Liwen,Mao Yilin,Zhao Xiufang,Wang Changhai.Isolation,purification and analysis of antialgal activities of antialgal com-pounds from Laminaria japonica[J].Acta Oceanologica Sinica (in Chinese),2022,44(2):143-154.
Authors:Sun Yingying  Zhu Wenxuan  Zhou Jing  Zhuang Liwen  Mao Yilin  Zhao Xiufang  Wang Changhai
Affiliation:(Jiangsu Provincial Key Laboratory of Marine Biotechnology,Jiangsu Ocean University,Lianyungang 222005,China;Co-Innovation Center of Jiangsu Marine Bio-industry Technology,Jiangsu Ocean University,Lianyungang 222005,China;Jiangsu Institute of Marine Resources Development,Lianyungang 222005,China;Lianyungang Quality and Technology Comprehensive Inspection and Testing Cen-ter,Lianyungang 222005,China;College of Resources and Environment Sciences,Nanjing Agricultural University,Nanjing 210095,China)
Abstract:Using 2 000 g kelp as raw material and Karenia mikimotoi as the test microalgae, methanol extraction, liquid-liquid extraction, silica gel column chromatography, sephadax LH-20 gel column chromatography and silica gel thin layer chromatography are used to purify compounds that might inhibit algal activity. Five samples with thin layer purity, B21 (1.39 g), B131 (0.51 g), B132 (0.48 g), C211 (0.016 g) and C212 (0.020 g), are prepared. Structural identification shows that B21, B131, B132, C211 and C212 are 7-hydroxyheptanic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, and oleic acid, respectively. They are isolated from Laminaria japonica for the first time. The results show that these five compounds have selective inhibition effects on the growth of six species of red tide microalgae (Alexandrium tamarense, Amphidinium carterae, Heterosigma akashiwo, Karenia mikimotoi, Phaeocystis globsa, and Skeletonema costatum). Among them, 7-hydroxyheptanoic acid has a wide range of antialgal activities. Except for Alexandrium tamarense, at the concentration of 50 μg/mL, the growth inhibition of 7-hydroxyheptanic acid against the other five species of red tide microalgae is close to that of copper sulfate and potassium dichromate, and the growth inhibition is more than 71% (the 4th day). Tetradecanoic acid also shows obvious inhibition on the growth of Amphidinium carterae, Karenia mikimotoi, and Skeletonema costatum. On 4th day, the growth inhibition of tetradecanoic acid for the three species of red tide microalgae is more than 57%. Furthermore, EC50?96 h values of 7-hydroxyheptanic acid, tetradecanoic acid, hexadecanoic acid, and octadecanoic acid for Amphidinium carterae, Heterosigma akashiwo, Karenia mikimotoi, Phaeocystis globsa, and Skeletonema costatum are obtained for the first time, respectively. EC50?96 h value of 7-hydroxyheptanic acid for Alexandrium tamarense is also determined for the first time. It is found that 7-hydroxyheptanic acid and tetradecanoic acid has the advantage of inhibiting Amphidinium carterae.
Keywords:antialgal compound  isolation  Laminaria japonica  purification  red tide microalgae
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