脂肪酸Bacillus pumilus羟基化调控基因fadD-like的克隆与序列分析

利用短小芽孢杆菌(Bacillus pumilus)的ω-1-羟基脂肪酸高产突变菌株M-F641、M-F81、M-F8272及B.pumilus20075的总DNA为模板,通过Primer Premier5.0软件设计引物,对决定长链脂肪酸无效降解途径中的关键酶基因fadD-like基因进行克隆,得到4条长约1 720 bp的fadD-like基因全序列;利用MEGA3.1、DNAStar等软件进行序列分析结合脂酰辅酶A合成酶系统进化分析,结果表明其与fadD基因具有较高的相似性,克隆得到的核苷酸为编码脂酰辅酶A合成酶的fadD基因序列;研究结果为长链脂肪酸高效转化生产ω-1-羟基脂肪酸提供基础。

Cloning and Sequence Analysis of fadD-like Gene for Metabolic Regulation in Hydroxy Fatty Acid Production with Bacillus pumilus

Using chromosomal DNA of ω-1-hydroxy fatty acids (HFA) high-yielding mutant Bacillus pumilus strains M-F641, M-F81, M-F8272 and B. pumilus 20075 as a template, the gene of deciding longchain fatty acid degradation was amplified by the PCR method and the primer was designed using software Primer Premier 5.0. Meanwhile, four full-length sequences of fadD-like gene about 1720 bp were obtained, and were researched by using software MEGA 3.1 DNAStar for sequence analysis and by acyl-coenzyme A synthetase for phylogenetic analysis. Results show that the sequence of the fragment from the four B. pumilus strains has high similarity with the fadD gene reported in reference, and the cloned nucleotide may have a gene sequence of the fadD encoding acyl-coenzyme A synthetase. This result provides the base for bioeonversion of long-chain fatty acids for ω-1-HFA production with high efficiency.