Diep D B, Håvarstein L S, Nes I F
Department of Biotechnological Sciences, Agricultural University of Norway, As, Norway.
Mol Microbiol. 1995 Nov;18(4):631-9. doi: 10.1111/j.1365-2958.1995.mmi_18040631.x.
In this study, we show that bacteriocin production in Lactobacillus plantarum C11 is an inducible process triggered by a secreted protein factor produced by the bacteriocin producer itself. The induction factor was identified to be plantaricin A, a bacteriocin-like peptide whose gene (plnA) is located in the same operon as a two-component regulatory system (plnBCD). When L. plantarum C11 cultures were depleted for plantaricin A, either by growing individual colonies on agar plates or by starting a new culture with a highly diluted inoculum, no bacteriocin was produced during the following growth. When chemically synthesized plantaricin A or purified bacterially produced plantaricin A was added to non-producing cultures, bacteriocin production was induced. Only 1 ng ml-1 plantaricin A is sufficient to induce the bacteriocin production in non-producing L. plantarum C11, and bacteriocin activity appears in the growth medium approximately 150 min after induction. Northern analyses, using a plnA-specific probe, demonstrated that plantaricin A is able to induce its own synthesis by transcription of the plnABCD operon, and this is observed approximately 15 min after adding plantaricin A. Furthermore, heterologous expression of the plnABCD operon in a Lactobacillus sake strain showed that the conditioned growth medium contained the active induction factor. Neither synthetic nor expressed plantaricin A from the heterologous system possesses any bacteriocin activity, suggesting that plantaricin A is primarily an induction factor and not a bacteriocin as claimed earlier.
在本研究中,我们发现植物乳杆菌C11中细菌素的产生是一个由细菌素产生菌自身分泌的蛋白质因子触发的诱导过程。诱导因子被鉴定为植物乳杆菌素A,一种类似细菌素的肽,其基因(plnA)与双组分调节系统(plnBCD)位于同一个操纵子中。当植物乳杆菌C11培养物中植物乳杆菌素A耗尽时,无论是通过在琼脂平板上培养单个菌落,还是通过用高度稀释的接种物启动新的培养物,在随后的生长过程中都不会产生细菌素。当将化学合成的植物乳杆菌素A或纯化的细菌产生的植物乳杆菌素A添加到不产生细菌素的培养物中时,细菌素的产生被诱导。仅1 ng/ml的植物乳杆菌素A就足以诱导不产生细菌素的植物乳杆菌C11产生细菌素,并且细菌素活性在诱导后约150分钟出现在生长培养基中。使用plnA特异性探针的Northern分析表明,植物乳杆菌素A能够通过plnABCD操纵子的转录诱导其自身的合成,并且在添加植物乳杆菌素A后约15分钟观察到这种情况。此外,plnABCD操纵子在清酒乳杆菌菌株中的异源表达表明,条件生长培养基中含有活性诱导因子。来自异源系统的合成或表达的植物乳杆菌素A均不具有任何细菌素活性,这表明植物乳杆菌素A主要是一种诱导因子,而不是如先前所声称的细菌素。