| Han Xiaotian,Wang Shuai,Zheng Li,Liu Wanshun. 2019. Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana. Acta Oceanologica Sinica, 38(2):107－113 |
| Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana |
| 球等鞭金藻中Δ12脂肪酸去饱和酶基因的克隆与功能鉴定 |
| Received:November 23, 2017 |
| DOI：10.1007/s13131-019-1354-1 |
| Key words:delta-12 fatty acid desaturase expression analysis Isochrysis galbana |
| 中文关键词: Δ12脂肪酸去饱和酶 表达分析 球等鞭金藻 |
| 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. |
| Author Name | Affiliation | E-mail | | Han Xiaotian | College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China
Changjiang Estuary Marine Ecosystem Research Station, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
Marine Ecology and Environmental Science Laboratory, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266237, China | | | Wang Shuai | Key Laboratory for Marine Bioactive Substances and Modern Analytical Technology, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China | | | Zheng Li | Marine Ecology and Environmental Science Laboratory, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266237, China
Key Laboratory for Marine Bioactive Substances and Modern Analytical Technology, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China | zhengli@fio.org.cn | | Liu Wanshun | College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China | |
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| Abstract: |
| 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. |
| 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。 |
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