| Han Xiaotian,Wang Shuai,Zheng Li,Liu Wanshun.Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana[J].Acta Oceanologica Sinica,2019,38(2):107-113 |
| 球等鞭金藻中Δ12脂肪酸去饱和酶基因的克隆与功能鉴定 |
| Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana |
| 投稿时间：2017-11-23 |
| DOI：10.1007/s13131-019-1354-1 |
| 中文关键词: Δ12脂肪酸去饱和酶 表达分析 球等鞭金藻 |
| 英文关键词:delta-12 fatty acid desaturase expression analysis Isochrysis galbana |
| 基金项目:The Basic Scientific Fund for National Public Research Institutes of China under contract No. 2017Q09; the Aoshan Science and Technology Innovation Project of Pilot National Laboratory for Marine Science and Technology (Qingdao) under contract No. 2016ASKJ02; the National Natural Science Foundation of China-Shandong Joint Funded Project under contract No. U1606404; the 973 Project from Chinese Ministry of Science and Technology under contract No. 2015CB755904; the National Natural Science Foundation of China under contract Nos 41776176 and 41806201; the Shandong Provincial Natural Science Foundation under contract No. ZR2015PD003. |
| 作者 | 单位 | E-mail | | 韩笑天 | 海洋生命学院, 中国海洋大学, 青岛, 266003, 中国
长江口海洋生态系统研究站, 中国科学院海洋研究所, 青岛, 266071, 中国
海洋生态与环境科学功能实验室, 青岛海洋科学与技术试点国家实验室, 青岛, 266237, 中国 | | | 王帅 | 海洋活性物质与现代分析技术重点实验室, 自然资源部第一海洋研究所, 青岛, 266061, 中国 | | | 郑立 | 海洋生态与环境科学功能实验室, 青岛海洋科学与技术试点国家实验室, 青岛, 266237, 中国
海洋活性物质与现代分析技术重点实验室, 自然资源部第一海洋研究所, 青岛, 266061, 中国 | zhengli@fio.org.cn | | 刘万顺 | 海洋生命学院, 中国海洋大学, 青岛, 266003, 中国 | |
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| 中文摘要: |
| 球等鞭金藻（Isochrysis galbana）是一类富含二十二碳六烯酸的海洋单细胞微藻。本研究利用SMART RACE技术成功克隆了球等鞭金藻的Δ12脂肪酸去饱和酶基因全长序列（IgFAD2），其开放阅读框（ORF）全长1，158 bp，编码386个氨基酸，相对分子质量为42.80 kDa，等电点为9.20。蛋白序列分析表明，含有3个高度保守的组氨酸簇并具有4个明显的跨膜α螺旋区，具有脂肪酸去饱和酶家族的典型特点。荧光实时定量PCR分析表明，在低温（15℃）、高盐（62‰和93‰）和N限制（220 μmol/L）条件下IgFAD2基因表现为上调表达。异源表达分析表明，含有IgFAD2基因表达载体的酵母菌株积累了C18：2Δ9，12。该研究结果表明球等鞭金藻的Δ12脂肪酸去饱和酶基因在不饱和脂肪酸合成代谢过程中具有重要作用。 |
| 英文摘要: |
| The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open reading frame measured a length of 1 158 bp and encoded 386 amino acids with a predicted molecular weight of 42.8 kDa and an isoelectric point of 9.2. Computational analysis of the protein sequence of IgFAD2 showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane-spanning regions that were universally present among plant desaturases. Quantitative real-time PCR results showed that the abundance of IgFAD2 transcript was significantly upregulated under different environmental stresses including low temperature (15℃), high salinity (salinity of 62 and 93), and nitrogen starvation (220 μmol/L). Heterologous expression indicated that yeast cells transformed with a plasmid construct containing IgFAD2 could convert endogenous oleic acid (18:1Δ9, OA) into linoleic acid (18:2Δ9, 12, LA). These findings confirm that I. galbana IgFAD2 plays important roles in the biosynthetic pathways of unsaturated fatty acids. |
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