Involvement of acyl-CoA synthetase genes in n-alkane assimilation and fatty acid utilization in yeast Yarrowia lipolytica
Involvement of acyl-CoA synthetase genes in n-alkane assimilation and fatty acid utilization in yeast Yarrowia lipolytica
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解脂耶氏酵母中酰基辅酶A合成酶基因参与正烷烃同化和脂肪酸利用
DOI：
10.1093/femsyr/fov031
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发表时间：
2015
影响因子：
3.2
通讯作者：
Ryouichi Fukuda
中科院分区：
文献类型：
--
作者：
Tenagy;Jun Seok Park;Ryo Iwama;Satoshi Kobayashi;Akinori Ohta;Hiroyuki Horiuchi;Ryouichi Fukuda
本文研究了YAL 1（FAA 1）和FAT 1编码的酰基辅酶A合成酶（ACS）以及另外三个ACS同源基因FAT 2-FAT 4在Yarrowia lipolytica正构烷烃和脂肪酸同化或利用中的作用。产生ACS缺失突变体以表征其功能。FAT 1缺失突变体在10-18个碳原子的正构烷烃上的生长减少，而FAA 1突变体在16个碳原子的正构烷烃上的生长减少。然而，FAT 2-FAT 4缺失突变体没有表现出任何生长缺陷，这表明FAT 1和FAA 1参与了正构烷烃代谢过程中产生的脂肪酸的活化。与此相反，FAA 1和FAT 1-FAT 4的缺失对脂肪酸的生长没有影响。野生型菌株在脂肪酸合成抑制剂浅蓝菌素的存在下生长，通过利用外源添加的脂肪酸或正烷烃衍生的脂肪酸，同时补充油酸或18个碳的正烷烃。然而，FAA 1缺失突变体没有生长，表明FAA 1在脂肪酸利用中起关键作用。荧光显微镜观察和生化分析表明，Fat 1 p是存在于过氧化物酶体和Faa 1 p是本地化的细胞质和膜。
Here, we investigated the roles ofYAL1(FAA1) andFAT1encoding acyl-CoA synthetases (ACSs) and three additional orthologs of ACS genesFAT2–FAT4of the yeastYarrowia lipolyticain the assimilation or utilization ofn-alkanes and fatty acids. ACS deletion mutants were generated to characterize their function. TheFAT1deletion mutant exhibited decreased growth onn-alkanes of 10–18 carbons, whereas theFAA1mutant showed growth reduction onn-alkane of 16 carbons. However,FAT2–FAT4deletion mutants did not show any growth defects, suggesting thatFAT1andFAA1are involved in the activation of fatty acids produced during the metabolism ofn-alkanes. In contrast, deletions ofFAA1andFAT1–FAT4conferred no defect in growth on fatty acids. The wild-type strain grew in the presence of cerulenin, an inhibitor of fatty acid synthesis, by utilizing exogenously added fatty acid or fatty acid derived fromn-alkane when oleic acid orn-alkane of 18 carbons was supplemented. However, theFAA1deletion mutant did not grow, indicating a critical role forFAA1in the utilization of fatty acids. Fluorescent microscopic observation and biochemical analyses suggested that Fat1p is present in the peroxisome and Faa1p is localized in the cytosol and to membranes.