Rawadi G, Lalanne J L, Roulland-Dussoix D
Centre de Recherche Roussel Uclaf, Romainville, France.
Gene. 1995 May 26;158(1):107-11. doi: 10.1016/0378-1119(95)00160-8.
Lipases, serine esterase enzymes, play an essential role in the mycoplasmal nutritional requirement for long-chain fatty acids. Although the lipase(s) activity in different mycoplasma species has been investigated, the molecular biology of the corresponding genes has not been studied. Using a single-primer PCR technique combined to more classical cloning systems, an operon containing three open reading frames (ORF), each of which could encode a lipase protein of 264, 264 or 269 amino acids (aa), was identified from Mycoplasma mycoides subsp. mycoides LC. Analysis of aa sequences of the encoded polypeptides showed that they display high aa similarity between each other (65-79%) and 28-31% identity to other prokaryotic lipases. Moreover, a lipase-esterase activity could be detected when the mycoplasmal lipase-encoding genes were expressed in a strong opal-suppressor-bearing Escherichia coli strain.
脂肪酶作为丝氨酸酯酶，在支原体对长链脂肪酸的营养需求中起着至关重要的作用。尽管已经对不同支原体物种中的脂肪酶活性进行了研究，但相应基因的分子生物学尚未得到研究。利用单引物PCR技术结合更经典的克隆系统，从丝状支原体丝状亚种LC中鉴定出一个含有三个开放阅读框（ORF）的操纵子，每个开放阅读框都可以编码一个由264、264或269个氨基酸（aa）组成的脂肪酶蛋白。对编码多肽的氨基酸序列分析表明，它们彼此之间具有很高的氨基酸相似性（65-79%），与其他原核脂肪酶的同一性为28-31%。此外，当支原体脂肪酶编码基因在携带强乳白抑制子的大肠杆菌菌株中表达时，可以检测到脂肪酶-酯酶活性。
