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Production and Ecological Significance of Yeast Cell Wall-Degrading Enzymes from Oerskovia

机译:Oerskovia酵母细胞壁降解酶的生产及其生态意义

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摘要

Motile actinomycetes capable of degrading walls of viable yeast cells were isolated from soil and identified as Oerskovia xanthineolytica. A lytic assay based on susceptibility of enzyme-treated cells to osmotic shock was developed, and 10 of 15 strains of O. xanthineolytica, Oerskovia turbata, and nonmotile Oerskovia- like organisms from other collections were found to possess yeast lytic activities. All lytic strains produced laminaranase and α-mannanase, but the amounts, determined by reducing group assays, were not proportional to the observed lytic activities. The Oerskovia isolates demonstrated chemotactic, predatory activity against various yeast strains and killed yeasts in mixed cultures. Of 15 carbon sources tested for production of lytic enzyme, purified yeast cell walls elicited the highest activity. Glucose repressed enzyme production and caused cells to remain in the microfilamentous and motile rod stages of the Oerskovia cell cycle. Crude lytic activity was optimal at pH 5.6 to 7.0 and inactivated by heating for 6 min at 50°C. Partial purification by isoelectric focusing showed that all lytic activity was associated with four β-(1→3)-glucanases. The absence of protein disulfide reductase, N-acetyl-β-d-hexosaminidase, and phosphomannanase in crude preparations indicated that the principal enzyme responsible for yeast wall lysis was a β-(1→3)-glucanase that produced relatively little reducing sugar from yeast glucan.
机译:从土壤中分离出能降解活酵母细胞壁的能动放线菌并鉴定为解毒欧氏菌。开发了一种基于酶处理的细胞对渗透性休克的敏感性的溶解测定法,发现其他菌株的15种O.xanthineolytica,Oerskovia turbata和非运动性Oerskovia样生物中的10株具有酵母裂解活性。所有裂解菌株均产生laminaranase和α-甘露聚糖酶,但通过还原基团测定确定的量与观察到的裂解活性不成比例。 Oerskovia分离物在混合培养物中表现出对多种酵母菌株的趋化,掠食性活性和杀死的酵母。在测试产生裂解酶的15种碳源中,纯化的酵母细胞壁具有最高的活性。葡萄糖抑制了酶的产生,并导致细胞停留在Oerskovia细胞周期的微丝和能动杆阶段。粗分解活性在pH 5.6至7.0时最佳,并在50°C加热6分钟使其失活。等电聚焦部分纯化表明所有裂解活性与四个β-(1→3)-葡聚糖酶有关。粗制品中不存在蛋白质二硫键还原酶,N-乙酰基-β-d-己糖胺酶和磷酸甘露聚糖酶,这表明负责酵母壁裂解的主要酶是β-(1→3)-葡聚糖酶,该酶从酵母中产生的还原糖相对较少酵母葡聚糖。

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