芥子碱和硫酸铜对芥子碱降解菌酶活力的影响

Effect of Sinapine and Copper Sulfate on Enzymes Activity of Sinapine-degrading Bacteria

【目的】本试验通过研究芥子碱、硫酸铜以及培养时间对芥子碱降解菌1(Sinapine-degrading Bacteria 1,SDB1)相关酶活力的影响,旨在探索SDB1中参与芥子碱降解的酶类以及影响酶活的因素。【方法】把30个SDB1的基础平板培养基平均分成1个对照组和4个处理组,每组6个重复。对照组除基础培养基外不再添加其他成分;处理1组和处理2组分别在基础培养基中添加0.40和0.80 g/L的芥子碱;处理3组和处理4组分别在基础培养基中添加1.00和3.00 mmol/L的硫酸铜。收集培养液,测定培养2、4、6、8 d后菌液中淀粉酶、多酚氧化酶、β-葡萄糖苷酶、漆酶、乙酰胆碱酯酶和脂肪酶的活力。【结果】①从对照组的结果看,SDB1具有一定的淀粉酶活性。与对照组相比,添加0.40 g/L芥子碱培养8 d,SDB1淀粉酶的活力是对照组的2.32倍(P<0.05),添加0.80 g/L芥子碱对SDB1淀粉酶活力没有显著影响。②SDB1具有多酚氧化酶活性。添加0.80 g/L芥子碱,多酚氧化酶活性在培养的第2和4天,分别比对照组增加了75.70%(P<0.01)和197.03%(P<0.01)。③SDB1具有较高的漆酶活性,且漆酶活性有随培养时间延长而增加的趋势。芥子碱可以提高SDB1的漆酶活力。添加0.40和0.80 g/L的芥子碱均可以提高SDB1漆酶活力,其中添加0.40 g/L的芥子碱培养4 d,SDB1漆酶活力是对照组的6.25倍(P<0.01);添加0.80 g/L的芥子碱培养4、6和8 d,漆酶活力分别为对照组的5.92(P<0.01)、3.03(P<0.01)和5.70倍(P<0.05)。④SDB1不具有β-葡萄糖苷酶活性,添加芥子碱对SDB1β-葡萄糖苷酶活力也没有显著影响。⑤SDB1具有一定的乙酰胆碱酯酶活性。与对照组相比,添加0.80 g/L芥子碱培养6 d,SDB1的乙酰胆碱酯酶活性显著降低(P<0.05),添加0.4 g/L芥子碱对SDB1乙酰胆碱酯酶活性没有显著影响。⑥SDB1具有一定的脂肪酶活性。添加0.40、0.80 g/L的芥子碱对SDB1脂肪酶活性没有显著影响。⑦添加1.00和3.00 mmol/L的硫酸铜对SDB1淀粉酶、多酚氧化酶、漆酶、β-葡萄糖苷酶、乙酰胆碱酯酶和脂肪酶活力均没有显著影响。(结论)本试验条件下,SDB1具有淀粉酶、多酚氧化酶、漆酶、乙酰胆碱酯酶和脂肪酶的活性,不具有β-葡萄糖苷酶的活性。在培养基中添加芥子碱,可以在培养的4~8 d内不同程度的提高SDB1淀粉酶、多酚氧化酶、漆酶活性;添加芥子碱对SDB1的β-葡萄糖苷酶、乙酰胆碱酯酶、脂肪酶活性没有显著影响;添加硫酸铜对SDB1的淀粉酶、多酚氧化酶、漆酶、β-葡萄糖苷酶、乙酰胆碱酯酶和脂肪酶活性均没有显著影响。

【Objective】To explore the enzymatic property of SDB1(Sinapine-degrading Bacteria 1)in degrading sinapine,the present article studied the effect of sinapine and copper sulfate on the activity of some enzymes in SDB1.【Method】30 basal plate medium of SDB1 were randomly allocated into 1 control group and 4 treatment group with 6 replicates per group.The control group added nothing except for the basal culture medium.Treatment group 1 and 2 were added 0.40 and 0.80 g/L sinapine respectively and the treatment group 3 and 4 were added 1.00 and 3.00 mmol/L copper sulfate besides the basal culture medium,respectively.The culture solution was collected and the activities of the amylase,polyphenol oxidase,β-glucosaccharase,laccase,acetylcholin esterase,lipase at the 2 th,4 th,6 th,8 th culture day were measured.【Result】(i)SDB1 had the activity of amylase.Amylase activity of the 0.40g/L sinapine group was 2.32(P<0.05)times as that of the control group in the 8 th day.There was no significant difference of the amylase activity between the 0.80 g/L sinapine group and the control group.(ii)SDB1 had the activity of polyphenol oxidase.Polyphenol oxidase activity of the 0.80 g/L sinapine group was 75.70%(P<0.01)and 197.03%(P<0.01)higher than that of the control group in the 2 th day and 4 th day,respectively.(iii)SDB1 had the high activity of laccase,and the laccase activity showed an increasing trend with time.Sinapine could increase the laccase activity of SDB1.Laccase activity of the 0.40 g/L sinapine group was 6.25(P<0.01)times as that of the control group in the 4 th day and of the 0.80 g/L sinapine group was 5.92(P<0.01),3.03(P<0.01),5.70(P<0.05)times as that of the control group in the 4 th,6 th,8 th day,respectively.(iv)SDB1 almost had no activity ofβ-glucosaccharase.(v)SDB1 had some activity of acetylcholin esterase.Acetylcholin esterase activity of the 0.80 g/L sinapine group decreased significantly(P<0.05)than that of the con-trol group in the 6 th day.There was no significant difference for acetylcholin esterase activity of SDB1 between the 0.40 g/L sinapine group and the control group.(vi)SDB1 had some activity of lipase.There was no significant difference for lipase activity for SDB1 between the sinapine groups and the control group.(vii)Copper sulfate at the dose of 1.00 and 3.00 mmol/L had no significant effect on the activity of am-ylase,polyphenol oxidase,β-glucosaccharase,laccase,acetylcholin esterase and lipase in SDB1.【Conclusion】This study confirms that SDB1 has the activity of amylase,polyphenol oxidase,laccase,acetylcholin esterase and lipase and has not the activity ofβ-glucosaccharase.Adding sinapine in the basal culture medium of SDB1 can variously increase the activity of amylase,polyphenol oxidase and laccase in SDB1 from the 4 th day to the 8 th day.There is no significant effect of sinapine onβ-glucosaccharase,acetylcholin esterase and lipase in SDB1.There is no significant effect of copper sulfate on amylase,polyphenol oxidase,laccase,β-glucosaccharase,acetylcholin esterase and lipase in SDB1.