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从海洋欧文氏菌中生产细胞壁降解酶及其生态意义。

Production and ecological significance of yeast cell wall-degrading enzymes from oerskovia.

机构信息

Department of Biochemistry and Microbiology, Cook College, Rutgers University, New Brunswick, New Jersey 08903.

出版信息

Appl Environ Microbiol. 1978 Oct;36(4):594-605. doi: 10.1128/aem.36.4.594-605.1978.

DOI:10.1128/aem.36.4.594-605.1978
PMID:16345321
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC243097/
Abstract

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 alpha-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 degrees C. Partial purification by isoelectric focusing showed that all lytic activity was associated with four beta-(1-->3)-glucanases. The absence of protein disulfide reductase, N-acetyl-beta-d-hexosaminidase, and phosphomannanase in crude preparations indicated that the principal enzyme responsible for yeast wall lysis was a beta-(1-->3)-glucanase that produced relatively little reducing sugar from yeast glucan.

摘要

能够降解活酵母细胞壁的能动放线菌从土壤中分离出来,并被鉴定为黄嘌呤氧化不动杆菌。建立了一种基于酶处理细胞对渗透冲击敏感性的裂解测定法,发现来自其他来源的 15 株黄嘌呤氧化不动杆菌、 turbata 不动杆菌和非能动黄嘌呤氧化不动杆菌中的 10 株具有酵母裂解活性。所有裂解菌株均产生纤维二糖酶和α-甘露聚糖酶,但通过还原基团测定法确定的量与观察到的裂解活性不成比例。Oerskovia 分离株对各种酵母菌株表现出趋化性和捕食活性,并在混合培养物中杀死酵母。在测试的 15 种碳源中,用于产生裂解酶的酵母细胞壁纯化产物引起了最高的活性。葡萄糖抑制酶的产生并导致细胞保持在 Oerskovia 细胞周期的微丝和能动杆状阶段。粗裂解活性在 pH5.6 至 7.0 时最佳,并在 50°C 加热 6 分钟失活。等电聚焦的部分纯化表明,所有裂解活性都与四种β-(1-->3)-葡聚糖酶有关。粗制剂中缺乏蛋白二硫键还原酶、N-乙酰-β-d-己糖胺酶和磷酸甘露聚糖酶表明,负责酵母细胞壁裂解的主要酶是β-(1-->3)-葡聚糖酶,它从酵母葡聚糖中产生相对较少的还原糖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e136/243097/2613e8336ee2/aem00213-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e136/243097/2613e8336ee2/aem00213-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e136/243097/2613e8336ee2/aem00213-0071-a.jpg

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本文引用的文献

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ENZYMATIC HYDROLYSIS OF YEAST CELL WALLS. I. ISOLATION OF WALL-DECOMPOSING ORGANISMS AND SEPARATION AND PURIFICATION OF LYTIC ENZYMES.酵母细胞壁的酶解。I. 壁分解微生物的分离以及裂解酶的分离与纯化。
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