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土壤拮抗链霉菌R_(15)β-1,3-葡聚糖酶的纯化与部分特性分析

Keywords: 链霉菌,β-1,3-葡聚糖酶,纯化,特性

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Abstract:

拮抗链霉菌(Steptomycesspp.)R15菌株是从大白菜植株根际土壤分离得到的,其代谢产生的抗菌物质对多种植物病原真菌有很强的抑制作用。采用ABP选择性培养基诱导培养法,获得4株(R8,R15,R31和AS)产β-1,3-葡聚糖酶菌株。其中链霉菌R15菌株在ABP平板上培养7d后,透明圈最大(>12mm),澄清度最高。从R15菌株提取酶液,经10% ̄75%的硫酸铵沉淀盐析、羟基磷灰石柱层析及DEAE-52柱层析得到纯化,纯化倍数为13.98,回收率达4.031%。经SDS-聚丙烯酰胺凝胶电泳测得该酶的分子量为41.3kDa。酶特性研究结果表明,该酶作用的最适温度为55℃,最适pH为5.0,低于55℃、pH7.5以下酶较稳定。Fe3+、K+、Cu2+、Hg2+对酶有抑制作用,而Ca2+、Fe2+、Ba2+、Mn2+、Al3+及Zn2+对酶有激活作用。

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