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念珠藻活性物质筛选及培养条件对其含量影响的研究
Alternative TitleTHE SCREENING OF BIOACTIVE COMPOUNDS FROM NOSTOC AND THE IMPACT OF GROWTH CONDITIONS ON ITS TYROSINASE INHIBITOR PRODUCTION
范羽仪
Subtype硕士
Thesis Advisor胡征宇
2008-06-17
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword念珠藻 代谢产物 生物活性 培养条件 中心组合设计
Abstract念珠藻属(Nostoc)是广泛分布于世界的一类丝状蓝藻,其次级代谢产物中有许多化合物以前未被发现。其中一些成分由于具有抗病毒,抗肿瘤等生物活性,引起了人们对它们进行开发和利用的极大兴趣。本文对十株念珠藻的活性物质进行了酪氨酸酶抑制剂,胰蛋白酶抑制剂等筛选,并研究了不同培养条件对其活性物质含量的影响,为今后进一步开发利用这些活性物质提供了参考依据。 在对十株念珠藻的提取物进行酪氨酸酶抑制剂,胰蛋白酶抑制剂等活性物质筛选时,发现十株念珠藻中有六株具有抑制酪氨酸酶活性的能力,这六株分别为点形念珠藻FACHB252,喜钙念珠藻FACHB389,胶团念珠藻FACHB392,以及三株未定种念珠藻(FACHB95,FACHB106和FACHB892);其甲醇提取物对酪氨酸酶抑制效果用IC50表示分别为33.32,29.67,41.32,43.21,43.95及23.92 µg/mL。有五株具有抑制胰蛋白酶活性的能力,这五株分别是沼泽念珠藻FACHB89,喜钙念珠藻FACHB389,胶团念珠藻FACHB392以及两株未定种念珠藻(FACHB106和FACHB139)。对胰蛋白酶活性的抑制作用比对酪氨酸酶活性的抑制作用要弱一些。同时,在十株念珠藻中,有七株影响超氧阴离子自由基的产生,它们分别是沼泽念珠藻FACHB89,林氏念珠FACHB103,点形念珠藻FACHB252,喜钙念珠藻FACHB389,胶团念珠藻FACHB392和及两株未定种念珠藻(FACHB95和FACHB892)。 比较自养条件和混合营养对喜钙念珠藻的生长发现,在BG-11培养基中加入一定含量的葡萄糖对酪氨酸酶抑制剂含量的提高有一定的帮助。在四组不同培养条件中,含有5 mM葡萄糖的BG-11培养基培养出的喜钙念珠藻,其提取物对酪氨酸酶活性的抑制效果最强。同时还发现,喜钙念珠藻提取物在33.3 µg/mL时对酪氨酸酶活性的抑制率与其细胞内可溶性有机碳含量成正相关。通过中心组合设计,比较不同光强和磷浓度对喜钙念珠藻生长的影响,发现磷浓度和光强对酪氨酸酶抑制剂在喜钙念珠藻中的产生没有交互作用。酪氨酸酶抑制剂在喜钙念珠藻细胞中的含量是随培养时间变化的,高磷和适当的光强(磷浓度70 mg/L,光强30 µmol• m-2• s-1)对细胞生长平台期酪氨酸酶抑制剂的积累具有很大的促进作用。
Other AbstractNostoc, as cyanobacteria in gereral, are widely distributed on earth. Many secondary metabolites with new structures have been isolated from these organisms. Due to their value of antiviral and antitumor activity, etc., the development and utilization of these bioactive compounds have gained much interest. This research screened secondary metabolites with tyrosinase inhibitory activity and trypsin inhibitory activity, etc. from 10 strains of Nostoc. In addition, the effects of different growth conditions (nutritional factors and environmental factor) on the production of bioactive compounds are investigated as well for the future culture, further development and utilization of these bioactive compounds. Through the screening of tyrosinase inhibitor, trypsin inhibitor and other bioactivity, it was found that 6 strains of Nostoc exhibit tyrosinase inhibitory activity, which are N. sp. FACHB95, N. sp. FACHB106, N. punctiforme FACHB252, N. calcicola FACHB389, N. parmelioides FACHB392, and N. sp. FACHB892, with IC50 of 43.21, 43.95, 33.32, 29.67, 41.32 and 23.92 µg/mL respectively, and 5 strains of Nostoc own inhibitory activity on trypsin, which are N. paludosum FACHB89,N. sp. FACHB106,N. sp. FACHB139,N. calcicola FACHB389,and N. parmelioides FACHB392. In general, the inhibitory ability on trypsin are weaker than that of tyrosinase. Meanwhile, the extracts from 7 strains have positive or negative effects on the genesis of superoxide anion free radical, which are N. paludosum FACHB89, N. sp. FACHB95, N. linckia FACHB103, N. punctiforme FACHB252, N. calcicola FACHB389, N. parmelioides FACHB392 and N. sp. FACHB892。 By comparing the autotrophic and mixotrophic conditions, it could be concluded that BG-11 medium with 5 mM glucose help to increase the inhibitory activity on tyrosinase from the extracts of N. calcicola FACHB389. And at the concentration of 33.3 µg/mL, the inhibitory rate of extract of N. calcicola FACHB389 on tyrosinase has a positive correlation with the content of TOC. While investigating the effects of phosphate concentration and light intensity on the production of tyrosinase inhibitor from N. calcicola FACHB389 by central composite design, the results suggest that phosphate concentration and light intensity have no interaction effects on the production of tyrosinase inhibitor, the content of tyrosinase inhibitor change with culture time, high phosphate concentration (70 mg/mL) and optimum light intensity (30 µmol• m-2• s-1) help the accumulation of tyosinase inhibitor in N. calcicola FACHB389 during stationary phase.
Pages69
Language中文
Document Type学位论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/12336
Collection学位论文
Recommended Citation
GB/T 7714
范羽仪. 念珠藻活性物质筛选及培养条件对其含量影响的研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2008.
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