白腐真菌共培养对磺胺二甲基嘧啶降解的影响

doi:10.13560/j.cnki.biotech.bull.1985.2016-0905

摘要/Abstract

摘要： 磺胺类抗生素是全国检出频率较高的一种抗菌药物,目前对磺胺类抗生素的生物降解方面研究较少,而生物法具有无二次污染、绿色环保的优点。因此本实验对杂色云芝和黄孢原毛平革菌共培养降解磺胺二甲基嘧啶进行研究。采用高效液相色谱（HPLC）和紫外分光光度计分别分析抗生素的浓度与酶活。结果显示,平板对峙实验中两菌种间菌丝呈僵持状,种间区域漆酶酶活始终高于其他区域。两种白腐真菌液体混配4 d时可以获得具有较高漆酶酶活的粗酶液,该粗酶液在48 h内对磺胺二甲基嘧啶的降解率为25.4%,明显高于杂色云芝纯培养粗酶液的降解率（9.6%）。在加入天然助氧剂和人工助氧剂2,2'-联氨双（3-乙基苯并噻唑啉-6-磺酸）二铵盐（简称ABTS）后,混配粗酶液的降解率分别增至49%和93.1%。种间作用可以通过增加漆酶酶活以及产生助氧剂提升抗生素降解率,因此,共培养的白腐真菌可以作为抗生素污染生物治理的主要手段之一。

关键词: 白腐真菌, 共培养, 漆酶酶活, 磺胺二甲基嘧啶, 降解

Abstract: Sulfonamides are a class of antimicrobial agents that have been detected in China at high-frequency. Biological method has the advantages of no secondary pollution and green environmental protection,but the researches on biodegradation of sulfonamides is still scarce. This experiment was to investigate degradation of sulfamethazine（SM2）by co-culturing Trametes versicolor and Phanerochaete chrysosporium. Antibiotic concentration and enzyme activity were determined using high performance liquid chromatography（HPLC）and UV spectrophotometer. A deadlock between the two mycelia was observed on plate confrontation method of an agar medium. The laccase activity at the interaction zone was higher than that at the other regions in the incubation of the two white-rot fungi on an agar plate. The crude enzyme preparation with high laccase concentration was obtained at the co-culturing time of 4 days in liquid medium. At 48 h after incubation,the degradation percentage of SM2 by crude enzyme solution from co-culturing was 25.4%,which was higher than that by crude enzyme preparations from Trametes versicolor（9.6%）. When natural mediators and artificial mediator 2,2’-azinobis（3-ethylbenzthiazoline-6-sulfonic acid ammonium salt）（ABTS）were added to the reaction mixture,the degradation percentages of SM2 by the crude enzyme from mixed culture significantly increased to 49% and 93.1%,respectively. The results showed that the interspecific interactions involving P. chrysosporium and T. versicolor enhanced the SM2 oxidation through increasing laccase activity and producing potential mediators. Therefore,the co-culturing of different white-rot fungi has potential for application in antibiotic-contaminated soils and wastes.

Key words: white-rot fungi, co-culture, laccase activity, sulfamethazine, degradation

郭晓丹娜,郭夏丽. 白腐真菌共培养对磺胺二甲基嘧啶降解的影响[J]. 生物技术通报, 2017, 33(5): 197-202.

GUO Xiaodanna GUO Xia-li. Degradation of Sulfamethazine by Co-culturing of White-rot Fungi[J]. Biotechnology Bulletin, 2017, 33(5): 197-202.

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