摘要
采用茚三酮试剂显色的薄层原位化学反应和抑藻活性检测方法,从龙须菜、条斑紫菜、鹿角藻和裙带菜中筛选环肽,发现龙须菜和条斑紫菜中存在具有抑藻活性的环肽。以龙须菜和条斑紫菜粉末为原料,采用硅胶柱层析和(或)硅胶薄层层析制备等方法,从龙须菜中分离到环(脯氨酸-甘氨酸)、环(丝氨酸-脯氨酸)和环(丙氨酸-色氨酸),从条斑紫菜中纯化到环(亮氨酸-脯氨酸)和环(缬氨酸-酪氨酸)。除环(丝氨酸-脯氨酸)外,其余4种环二肽均是首次从相应大型海藻中获得。环(脯氨酸-甘氨酸)、环(亮氨酸-脯氨酸)和环(缬氨酸-酪氨酸)明显抑制了强壮前沟藻、赤潮异弯藻和球形棕囊藻的生长,对米氏凯伦藻和中肋骨条藻有选择性抑制作用。在50μg/mL时,3种环二肽对强壮前沟藻、赤潮异弯藻和球形棕囊藻的生长抑制率≥50%(第4d);环(脯氨酸-甘氨酸)对米氏凯伦藻以及环(亮氨酸-脯氨酸)对中肋骨条藻的生长抑制率超过或接近50%。随后,获得环二肽对赤潮微藻生长的半抑制效应浓度EC50-96h。结果表明,环(脯氨酸-甘氨酸)、环(亮氨酸-脯氨酸)和环(缬氨酸-酪氨酸)在抑制米氏凯伦藻、球形棕囊藻或赤潮异弯藻方面比重铬酸钾更有优势,有望开发为环境友好型赤潮微藻抑藻剂。
The cyclic peptides with antialgal activity were screened from Gracilaria lemaneiformis,Neoporphyra yezoensis,Silvetia siliguosa and Undaria pinnatifida,by thin-layer in-situ chemical reaction with ninhydrin reagent and antialgal activity test.The cyclic peptides with antialgal activity were found in the extracts of Gracilaria lemaneiformis and Neoporphyra yezoensis,respectively.Cyclo(Pro-Gly),cyclo(Ser-Pro),cyclo(Ala-Trp)were finally isolated from Gracilaria lemaneiformis,and cyclo(Leu-Pro)and cyclo(Val-Tyr)were purified from Neoporphyra yezoensis by repeated silica gel column chromatography and(or)silica gel thin layer chromatography.Except for cyclo(Ser-Pro),the other four cyclic dipeptides were obtained from corresponding marine macroalgae for the first time.Cyclo(Pro-Gly),cyclo(Leu-Pro)and cyclo(Val-Tyr)significantly inhibited the growth of Amphidinium carterae,Heterosigma akashiwo,and Phaeocystis globosa;they showed selective growth inhibition for Karenia mikimotoi and Skeletonema costatum.At the concentration of 50μg/mL,the growth inhibition of cyclo(Pro-Gly),cyclo(Leu-Pro)and cyclo(Val-Tyr)for Amphidinium carterae,Heterosigma akashiwo and Phaeocystis globosa was≥50%(on the 4th day);the growth inhibition of cyclo(Pro-Gly)for Karenia mikimotoi and that of cyclo(Leu-Pro)for Skeletonema costatum were more than or close to 50%.Further,EC_(50-96h)of cyclic dipeptide for red tide microalgae was obtained.EC_(50-96h)of cyclo(Pro-Gly)for Karenia mikimotoi and Phaeocystis globosa,EC_(50-96h)of cyclo(Leu-Pro)and cyclo(Val-Tyr)for Heterosigma akashiwo were less than that of potassium dichromate for corresponding red tide microalgae.This suggested that cyclo(Pro-Gly),cyclo(Leu-Pro)and cyclo(Val-Tyr)have more advantages in inhibiting Karenia mikimotoi,Phaeocystis globosa or Heterosigma akashiwo,and are expected to be developed into an environment-friendly algal inhibitor against red tide microalgae.
作者
庄丽雯
周文静
毛奕淋
郭赣林
王长海
孙颖颖
ZHUANG Liwen;ZHOU Wenjing;MAO Yilin;GUO Ganlin;WANG Changhai;SUN Yingying(Jiangsu Key Laboratory of Marine Biotechnology,Jiangsu Ocean University,Lianyungang 222005,China;Co-Innovation Center of Jiangsu Marine Bio-industry Technology,Lianyungang 222005,China;Jiangsu Key Laboratory of Marine Bioresources and Eco-environment,Jiangsu Ocean University,Lianyungang 222005,China;Resources and environment science institute,Nanjing Agricultural University,Nanjing 210095,China)
关键词
龙须菜
条斑紫菜
抑藻活性
环二肽
分离纯化
Gracilaria lemaneiformi
Neoporphyra yezoensis
antialgal activity
diketopiperazines
isolation
