Department of Food Science and Technology, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece.
Appl Environ Microbiol. 2010 Feb;76(4):1143-51. doi: 10.1128/AEM.00151-09. Epub 2009 Dec 18.
The aim of the present work was to study the mode of the induction of the biosynthesis of macedocin, the lantibiotic produced by Streptococcus macedonicus ACA-DC 198. Macedocin was produced when the strain was grown in milk but not in MRS or M17 broth. No autoinduction mechanism was observed. Production did not depend on the presence of lactose or galactose in the culture medium or on a coculture of the producer strain with macedocin-sensitive or macedocin-resistant strains. Induction seemed to depend on the presence of one or more heat-stable protein components produced when S. macedonicus ACA-DC 198 was grown in milk. The partial purification of the induction factor was performed by a combination of chromatography methods, and its activity was confirmed by a reverse transcription-PCR approach (RT-PCR). Mass spectrometric (MS) and tandem mass spectrometric (MS/MS) analyses of an induction-active fraction showed the presence of several peptides of low molecular mass corresponding to fragments of alpha(S1)- and beta-casein as well as beta-lactoglobulin. The chemically synthesized alpha(S1)-casein fragment 37-55 (2,253.65 Da) was proven to be able to induce macedocin biosynthesis. This is the first time that milk protein degradation fragments are reported to exhibit a bacteriocin induction activity.
本工作的目的是研究由马祖链霉菌 ACA-DC 198 产生的林可肽 macedocin 的生物合成诱导模式。当该菌株在牛奶中生长时会产生 macedocin,但在 MRS 或 M17 肉汤中不会产生。没有观察到自动诱导机制。生产不依赖于培养基中乳糖或半乳糖的存在,也不依赖于产菌株与 macedocin 敏感或 macedocin 抗性菌株的共培养。诱导似乎取决于马祖链霉菌 ACA-DC 198 在牛奶中生长时产生的一种或多种热稳定蛋白成分的存在。通过色谱方法的组合对诱导因子进行了部分纯化,并通过逆转录-PCR 方法(RT-PCR)证实了其活性。诱导活性部分的质谱(MS)和串联质谱(MS/MS)分析表明,存在几种低分子量的肽,对应于 alpha(S1)-和 beta-酪蛋白以及 beta-乳球蛋白的片段。经化学合成的 alpha(S1)-酪蛋白片段 37-55(2,253.65 Da)被证明能够诱导 macedocin 生物合成。这是首次报道乳蛋白降解片段具有细菌素诱导活性。