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首页> 外文期刊>Turkish journal of biology >Large-scale purification of a bacteriocin produced by Leuconostoc mesenteroides subsp. cremoris using diatomite calcium silicate
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Large-scale purification of a bacteriocin produced by Leuconostoc mesenteroides subsp. cremoris using diatomite calcium silicate

机译：大规模纯化肠膜亮白亚种产生的细菌素。使用硅藻土硅酸钙的cremoris

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Leuconostoc mesenteroides subsp. cremoris strain W3, isolated from wine, was previously found to produce a bacteriocin with inhibitory activity towards malolactic bacteria and hence inhibit malolactic fermentation in model wine medium. The bacteriocin, which we termed mesentericin W3, was purified by multistep chromatography and found in a previous study to be similar to mesentericin Y105, albeit with a low recovery (3.2%). In this study, we aimed to achieve large-scale isolation of mesentericin W3 by adsorption of the bacteriocin from culture supernatant onto Micro-Cel (diatomite calcium silicate) and then by subsequent desorption. Water-soluble surfactants including Tween 80, Triton X-100, sodium deoxycholate, sodium dodecyl sulfate (SDS) in distilled water, organic solvents (methanol, ethanol, acetonitrile), and distilled water with a pH range of 2–10 were tested for the desorption of the bacteriocin from Micro-Cel. Mesentericin W3 was successfully desorbed from Micro-Cel with SDS, while other tested reagents were not as successful. The bacteriocin desorbed from Micro-Cel showed the same inhibitory activity as the culture supernatant. The desorbed bacteriocin was applied to an SP Sepharose Fast Flow column for final purification. MALDI-TOF mass spectrometry confirmed the identity of the bacteriocin.
机译：肠膜明串珠菌亚种。以前发现从酒中分离出的cremoris菌株W3会产生一种细菌素，其对苹果酸乳酸菌具有抑制活性，因此可以在模型酒培养基中抑制苹果酸乳酸发酵。通过多步色谱法纯化了细菌素，我们称其为Mesentericin W3，在先前的研究中发现它与Mesentericin Y105相似，尽管回收率较低（3.2％）。在这项研究中，我们旨在通过将细菌素从培养上清液吸附到Micro-Cel（硅藻土硅酸钙）上，然后进行随后的解吸，来实现大规模分离Mesentericin W3。测试了水溶性表面活性剂，包括吐温80，Triton X-100，脱氧胆酸钠，蒸馏水中的十二烷基硫酸钠（SDS），有机溶剂（甲醇，乙醇，乙腈）和pH范围为2-10的蒸馏水。 Micro-Cel中细菌素的解吸。 Mesentericin W3已成功通过SDS从Micro-Cel上解吸，而其他测试试剂却不那么成功。从Micro-Cel解吸的细菌素显示出与培养上清液相同的抑制活性。将解吸的细菌素加到SP Sepharose Fast Flow色谱柱上进行最终纯化。 MALDI-TOF质谱法证实了细菌素的身份。

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