LsSpt23p is a regulator of triacylglycerol synthesis in the oleaginous yeast Lipomyces starkeyi
LsSpt23p is a regulator of triacylglycerol synthesis in the oleaginous yeast Lipomyces starkeyi
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LsSpt23p 是产油酵母斯达克油脂酵母中三酰甘油合成的调节剂
DOI：
10.1007/s00253-023-12361-2
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发表时间：
2023
影响因子：
5
通讯作者：
Aburatani Sachiyo
中科院分区：
文献类型：
--
作者：
Takaku Hiroaki;Kazama Haruka;Sato Rikako;Mori Kazuki;Ara Satoshi;Ishiya Koji;Matsuzawa Tomohiko;Yaoi Katsuro;Araki Hideo;Shida Yosuke;Ogasawara Wataru;Tashiro Kosuke;Kuhara Satoru;Yamazaki Harutake;Aburatani Sachiyo
产油酵母--斯氏油脂酵母（Lipomycesstarkeyi）在氮限制条件下能大量积累糖基化产物三酰甘油（TAG），具有很大的工业应用潜力。然而，L. starkeyi尚未得到深入调查。在这项研究中，我们比较了野生型和突变体的基因组序列增加TAG生产力，并确定了一个调节蛋白，LsSpt 23 p，这有助于调节TAG合成在L。starkeyi。L.与野生型菌株相比，过表达LsSPT 23的starkeymutants增加了TAG生产力。实时定量PCR分析表明，LsSpt 23 p上调了GPD 1（编码甘油3-磷酸脱氢酶）、肯尼迪途径基因SCT 1、SLC 1、PAH 1、DGA 1和DGA 2、柠檬酸介导的酰基辅酶A合成途径相关基因ACL 1、ACL 2、ACC 1、FAS 1和FAS 2以及OLE 1（编码β 9脂肪酸去饱和酶）的表达。染色质免疫沉淀-定量PCR分析表明LsSpt 23 p作为SLC 1和PAH 1，所有柠檬酸介导的酰基辅酶A合成途径相关基因和OLE 1的直接调节剂。这些结果表明LsSpt 23 p调节TAG合成。磷脂酸是磷脂酸磷酸水解酶（用于TAG合成）和磷脂酸胞苷酰转移酶1（用于Kennedy途径中的磷脂合成）的常见底物。LsSpt 23 p直接调控PAH 1的表达，但不影响CDS 1的表达，提示在氮限制条件下，碳的优先途径是Pah 1 p介导的TAG合成途径。本研究有助于了解TAG合成的调控机制，对提高产油酵母TAG产量具有重要意义。关键点LsSPT 23 p被鉴定为TAG生物合成的正调控因子LsSPT 23过表达增强TAG生物合成基因的表达和TAG产量LsSPT 23 M1108过表达突变体的TAG产量比对照高5倍。
The oleaginous yeastLipomyces starkeyihas considerable potential in industrial application, since it can accumulate a large amount of triacylglycerol (TAG), which is produced from sugars under nitrogen limitation condition. However, the regulation of lipogenesis inL. starkeyi has not been investigated in depth. In this study, we compared the genome sequences of wild-type and mutants with increased TAG productivity, and identified a regulatory protein, LsSpt23p, which contributes to the regulation of TAG synthesis inL. starkeyi. L. starkeyimutants overexpressingLsSPT23had increased TAG productivity compared with the wild-type strain. Quantitative real-time PCR analysis showed that LsSpt23p upregulated the expression ofGPD1, which encodes glycerol 3-phosphate dehydrogenase; the Kennedy pathway genesSCT1, SLC1, PAH1, DGA1, andDGA2; the citrate-mediated acyl-CoA synthesis pathway-related genesACL1, ACL2, ACC1, FAS1, and FAS2;andOLE1,which encodes ∆9 fatty acid desaturase. Chromatin immunoprecipitation-quantitative PCR assays indicated that LsSpt23p acts as a direct regulator ofSLC1andPAH1, all the citrate-mediated acyl-CoA synthesis pathway–related genes, andOLE1. These results indicate that LsSpt23p regulates TAG synthesis. Phosphatidic acid is a common substrate of phosphatidic acid phosphohydrolase, which is used for TAG synthesis, and phosphatidate cytidylyltransferase 1 for phospholipid synthesis in the Kennedy pathway. LsSpt23p directly regulatedPAH1 but did not affect the expression ofCDS1, suggesting that the preferred route of carbon is the Pah1p-mediated TAG synthesis pathway under nitrogen limitation condition. The present study contributes to understanding the regulation of TAG synthesis, and will be valuable in future improvement of TAG productivity in oleaginous yeasts.Key pointsLsSpt23p was identified as a positive regulator of TAG biosynthesisLsSPT23 overexpression enhanced TAG biosynthesis gene expression and TAG productionLsSPT23M1108Toverexpression mutant showed fivefold higher TAG production than controlGraphical abstract
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斯塔克脂肪酵母在不同条件下产生的脂肪成分
DOI：
10.1007/bf00406833
发表时间：
2004
期刊：
Archiv für Mikrobiologie
影响因子：
--
作者：
M. Deinema;C. Landheer
通讯作者：
C. Landheer
影响因子：
3.7
作者：
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通讯作者：
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影响因子：
9.9
作者：
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通讯作者：
Larroche, Christian
影响因子：
4.5
作者：
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通讯作者：
Takaku H
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影响因子：
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作者：
V. Makrantoni;Daniel Robertson;A. Marston
通讯作者：
A. Marston