Schjerling C K, Hummel R, Hansen J K, Borsting C, Mikkelsen J M, Kristiansen K, Knudsen J
Institute of Biochemistry, University of Odense, Campusvej 55, DK-5230 Odense M, Denmark.
J Biol Chem. 1996 Sep 13;271(37):22514-21. doi: 10.1074/jbc.271.37.22514.
The ACB1 gene encoding the acyl-CoA-binding protein (ACBP) was disrupted in Saccharomyces cerevisiae. The disruption did not affect the growth rate on glucose but reduced the growth rate on ethanol slightly. Although the growth rate of the acb1-disrupted cells was unaffected or only slightly affected, the acb1-disrupted strain was unable to compete with wild type cells when grown in mixed culture. The acyl-CoA level in the disrupted cells was increased from 1.5- to 2.5-fold during exponential growth. The increase in the acyl-CoA level was caused solely by an increase in de novo synthesized stearoyl-CoA. Experiments with purified yeast fatty acid synthetase show that it will synthesize long chain acyl-CoAs in the absence of acyl-CoA-binding protein. The addition of ACBP to the incubation medium resulted in a dramatic decrease in the chain length of the synthesized acyl-CoA esters. Despite the fact that the stearoyl-CoA concentration was increased 7-fold and the Delta9-desaturase mRNA level was increased 3-fold, the synthesis of oleic acid was unchanged in the acb1-disrupted strain. The results strongly indicate that ACBP in yeast is involved in the transport of newly synthesized acyl-CoA esters from the fatty acid synthetase to acyl-CoA-consuming processes.
编码酰基辅酶A结合蛋白(ACBP)的ACB1基因在酿酒酵母中被破坏。这种破坏不影响在葡萄糖上的生长速率,但略微降低了在乙醇上的生长速率。尽管acb1破坏细胞的生长速率未受影响或仅受到轻微影响,但在混合培养中生长时,acb1破坏菌株无法与野生型细胞竞争。在指数生长期间,破坏细胞中的酰基辅酶A水平增加了1.5至2.5倍。酰基辅酶A水平的增加完全是由新合成的硬脂酰辅酶A增加引起的。用纯化的酵母脂肪酸合成酶进行的实验表明,在没有酰基辅酶A结合蛋白的情况下,它会合成长链酰基辅酶A。向孵育培养基中添加ACBP导致合成的酰基辅酶A酯的链长显著降低。尽管硬脂酰辅酶A浓度增加了7倍,并且Delta9-去饱和酶mRNA水平增加了3倍,但在acb1破坏菌株中油酸的合成没有变化。结果强烈表明,酵母中的ACBP参与了新合成的酰基辅酶A酯从脂肪酸合成酶到酰基辅酶A消耗过程的转运。