丹参内生真菌产生淀粉酶菌株的筛选鉴定及酶学性质分析
Screening, identification of endophytic fungus produing raw starch-degrading enzyme from Salvia miltiorrhiza Bge.and its enzymatic properties
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摘要: 目的:从丹参内生真菌中筛选耐热性强的酸性生淀粉酶菌株,并对其酶学性质进行初步研究。方法:以50株丹参内生真菌作为筛选对象,通过淀粉固体培养基初筛、液体发酵复筛,筛选产生淀粉酶活力较高的菌株;通过形态学观察和ITS序列分析对产酶菌株进行鉴定。结果:从50株丹参内生真菌中,筛选出5株产生淀粉酶活力较高的菌株,其中菌株ZDH2-2-1-1为产淀粉酶活性最强的菌株,酶活力达到117.63 U/m L,经鉴定为互隔链格孢(Alternaria alternata)。该淀粉酶最适p H为4.0,最适反应温度为55℃,具有较好的热稳定性和p H稳定性,对生淀粉具有广谱的降解能力。通过薄层层析发现,ZDH2-2-1-1所产淀粉酶的酶解产物为葡萄糖。结论:丹参内生真菌能产生高活性的耐热性酸性生淀粉酶,在生淀粉深加工中具有极大的开发潜力。Abstract: Objective: Acid amylase producing endophytic fungus was screened from Salvia miltiorrhiza Bge. And its enzymatic properties were studied. Method: Amylase producing strains with high activity were screened by starch solid medium and fermentation medium, and identified by morphological observation and ITS sequence analysis. Results: 5 strains with higher amylase activity were screened from 50 endophytic fungi.Among them, the strain ZDH2-2-1-1 produced the highest activity of amylase, and the enzyme activity reached 117.63 U/m L, which was identified as Alternaria alternata. The enzyme properties showed that the optimal reaction p H of amylase from the strain ZDH2-2-1-1 was 4.0, the optimal reaction temperature was55 ℃, and the strain ZDH2-2-1-1 had good thermal stability, p H stability and a wide spectrum degradation ability of raw starch.By thin layer chromatography, the results showed that the enzymatic hydrolysis product of amylase produced by ZDH-2-2-1-1 was glucose.Conclusion: Endophytic fungi from Salvia miltiorrhiza Bge.can produce heat-resistant acid amylase, which has great development potential in the deep processing of raw starch.