一株扎布耶盐碱湖伪盐生杜氏藻的分离鉴定及生长影响因素分析  

Isolation,identification,and analysis of growth influencing factors of a Dunaliella pseudosalina strain from Zabuye Salt Lake

作　　者：郭敏 韩睿[2] 陶宇杰 高翔 邢江娃 王嵘 朱德锐 GUO Min;HAN Rui;TAO Yujie;GAO Xiang;XING Jiangwa;WANG Rong;ZHU Derui(Research Center for Basic Medical Science,Medical College,Qinghai University,Xining 810016,Qinghai,China;Key Laboratory of Vegetable Genetics and Physiology,Academy of Agriculture and Forestry Sciences,Qinghai University,Xining 810016,Qinghai,China)

机构地区：[1]青海大学医学院基础医学研究中心,青海西宁810016 [2]青海大学农林科学院蔬菜遗传与生理重点实验室,青海西宁810016

出　　处：《微生物学通报》2024年第11期4502-4516,共15页Microbiology China

基　　金：青海省基础应用研究计划(2022ZJ914)。

摘　　要：【背景】杜氏藻(Dunaliella)具有独特的生理特性和丰富的胞内次级代谢物,在医药、食品、养殖业、化工、轻工等领域具有广泛的应用价值。【目的】分析影响杜氏藻生物量的生长因素和最优培养条件,以期提高杜氏藻的生物量;明确最佳培养条件下杜氏藻胞内次级代谢物的种类。【方法】采用纯培养法分离伪盐生杜氏藻(Dunaliella pseudosalina)ZBY-1,18S rRNA基因测序明确其分类学地位;采用单因素试验探究其生长最佳条件以及营养盐种类及浓度;采用正交试验优化分离藻株的培养条件;利用液相色谱-质谱联用技术(LC-MS)检测藻株ZBY-1在最佳培养条件下的次级代谢产物。【结果】藻株ZBY-1的最适生长盐度为10%,最适生长pH值为8.5,最适生长温度为25℃。单因素分析显示最适碳源、氮源、磷源的种类和浓度分别为NaHCO_(3)1.26、CO(NH_(2))_(2)0.84、NaH_(2)PO_(4)0.06 g/L。优化生长条件后,藻株ZBY-1的细胞密度可达(2.57±0.12)×10^(7)个/mL,较优化前提高了1.25倍。LC-MS分析表明胞内次级代谢物的种类主要是萜类、生物碱类、苯丙素类、氨基酸相关化合物等;显著富集的代谢物通路是辅助因子生物合成、植物类次级代谢物合成和ABC转运等途径。【结论】扎布耶盐碱湖的地理位置特殊,藻类资源未被深入挖掘。分离藻株ZBY-1可适应高盐碱环境,为后续胞内次级代谢物的应用开发提供一定的参考依据。[Background]With rich metabolites and unique physiological properties,Dunaliella has wide applications in the pharmaceutical,food,aquaculture,chemical,and light industries.[Objective]To analyze the factors affecting the biomass of Dunaliella and the optimal culture conditions with a view to increasing the biomass of Dunaliella and identify the secondary metabolites of Dunaliella under the optimal culture conditions.[Methods]D.pseudosalina ZBY-1 was isolated by the culture method and identified by 18S rRNA gene sequencing.The growth conditions(nutrients and concentrations)were optimized by single factor and orthogonal experiments.The secondary metabolites of strain ZBY-1 cultured under the optimal conditions were then analyzed by LC-MS.[Results]The strain ZBY-1 showcased the best growth performance at 10%NaCl,pH 8.5,and 25℃.The optimal carbon,nitrogen,and phosphorus sources and their concentrations for this strain were NaHCO_(3)1.26 g/L,CO(NH_(2))_(2)0.84 g/L,and NaH_(2)PO_(4)0.06 g/L,respectively.The cell density of strain ZBY-1 cultured under the optimal conditions reached(2.57±0.12)×10^(7) cells/mL,which increased by 1.25 times compared with that before optimization.LC-MS results showed that secondary metabolites were mainly terpenoids,alkaloids,phenylpropanoids,and amino acid-related compounds.The enriched metabolite pathways were the cofactor biosynthesis pathway,plant-derived metabolite biosynthesis pathway,and the ABC transport pathway.[Conclusion]Zabuye Salt Lake has a special geographical location,whereas the algal resources remain to be exploited.The isolated strain ZBY-1 can adapt to the high saline-alkali environments,which provide a basis for subsequent development and application of the metabolites of this strain.

关 键 词：伪盐生杜氏藻 分离 生长特性 正交试验 代谢组学 

分 类 号：Q94-33[生物学—植物学]