| 磷酸泛酰巯基乙胺基转移酶基因过量表达对集胞藻PCC6803脂肪酸合成的影响 |
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| 引用本文： | 李燕乐,钟怀荣,宣宁,张燕,陈高,季祥.磷酸泛酰巯基乙胺基转移酶基因过量表达对集胞藻PCC6803脂肪酸合成的影响[J].浙江农业学报,2022,34(6):1316. |
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| 作者姓名： | 李燕乐 钟怀荣 宣宁 张燕 陈高 季祥 |
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| 作者单位： | 1.内蒙古科技大学 生命科学与技术学院,内蒙古自治区生物质能源化利用重点实验室,内蒙古 包头 0140102.山东省农业科学院 生物技术研究中心,山东省作物遗传改良与生态生理重点实验室,山东 济南 250100 |
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| 基金项目： | 内蒙古自然科学基金(2019MS02024);国家重点研发计划政府间国际科技创新合作/港澳台科技创新合作重点专项(2018YFE0108600);山东省重点研究计划（公益性科技攻关类）(2018GSF121019);山东省重点研究计划（公益性科技攻关类）(2019GSF107098) |
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| 摘 要： | 磷酸泛酰巯基乙胺基转移酶（phosphopantetheinyl transferases,PPTase）是细菌脂肪酸合成中的关键酶。本研究利用同源重组手段构建集胞藻PPTase基因（slr0495）过量表达重组质粒,在集胞藻PCC6803中进行表达研究,并在DNA和RNA水平进行验证,利用气相色谱-质谱联用仪（GC-MS）检测不同条件下突变藻株脂肪酸组分及含量。结果表明：在温度为30 ℃、光照强度为50 μmol ·m-2 · s-1培养条件下,过量表达slr0495基因突变藻株中C12:0、C16:0和C18:0的含量分别为1. 31 mg · g-1、8.07 mg · g-1和1.35 mg · g-1,比野生型藻株分别提高了23.58%、25.31%和13.45%;在温度为20 ℃、50% NaNO3浓度、光照强度为50 μmol · m-2 · s-1培养条件下,与野生型相比,突变藻株中C16:0和C18:0含量分别提高了44.71%和41.51%。以上结果表明,过表达slr0495基因增加了集胞藻PCC6803中长链饱和脂肪酸的含量,并且在低温（20 ℃）和缺氮（50% NaNO3）双重胁迫培养条件下中长链饱和脂肪酸含量得到进一步提高。本研究为探究slr0495基因功能以及在逆境中基因产生的应激反应提供了理论依据,为微藻高产多不饱和脂肪酸代谢研究奠定了基础。
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| 关 键 词： | 集胞藻PCC6803 磷酸泛酰巯基乙胺基转移酶 脂肪酸 同源重组 |
| 收稿时间： | 2021-01-29 |
Effects of over expression of phosphopantetheinyl transferases gene on fatty acid synthesis in Synechocystis sp. PCC6803 |
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| LI Yanle,ZHONG Huairong,XUAN Ning,ZHANG Yan,CHEN Gao,JI Xiang.Effects of over expression of phosphopantetheinyl transferases gene on fatty acid synthesis in Synechocystis sp. PCC6803[J].Acta Agriculturae Zhejiangensis,2022,34(6):1316. |
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| Authors: | LI Yanle ZHONG Huairong XUAN Ning ZHANG Yan CHEN Gao JI Xiang |
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| Institution: | 1. Inner Mongolia Key Laboratory of Biomass-Energy Conversion, School of Life and Technology, Inner Mongolia University of Science and Technology, Baotou 014010, Inner Mongolia, China
2. Shandong Key Laboratory of Crop Genetics and Breeding and Ecological Physiology, Biotechnology Research Center, Shandong Academy of Agricultural Sciences, Jinan 250100, China |
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| Abstract: | Phosphopantothenyl transferase (PPTase) is a key enzyme in fatty acid synthesis in bacteria. In this study, the homologous recombinant plasmid slr0495(+) was successfully constructed for overexpressing the PPTase (slr0495) gene and transformed into wild-type Synechocystis sp. PCC6803. The results showed that under the culture conditions of 30 ℃, 50 μmol·m-2·s-1, the contents of C12:0, C16:0 and C18: 0 in wild type Synechocystis sp. were 1.06 mg·g-1, 6.44 mg·g-1, 1.19 mg·g-1,while the C12:0, C16:0and C18:0 contents of mutant strains were 23.58%, 25.31% and 13.45% higher than those of wild type strains, respectively. Under the culture conditions of 20 ℃, 50% NaNO3 and 50 μmol·m-2·s-1 light intensity, compared with the wild type, the contents of C16:0 and C18:0 in mutant strains overexpressing slr0495 gene increased by 44.71% and 41.51%, respectively. These above results indicated that overexpression of slr0495 gene increased the contents of saturated long-chain fatty acids in Synechocystis sp. PCC6803, and the contents of medium-long-chain saturated fatty acids were further improved under the conditions of low temperature (20 ℃) and nitrogen deficiency (50% NaNO3). This study not only provides a theoretical basis for further exploring the function of slr0495 and the stress response of this gene in adversity but also lays the foundation for further exploring the high-yield polyunsaturated fatty acids of microalgae. |
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| Keywords: | Synechocystis sp PCC6803 phosphopantetheinyl transferases polyunsaturated fatty acids homologous recombination |
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