白腐真菌产漆酶培养基的优化及对嘧菌酯的降解

发布时间：2018-04-24 03:25

  本文选题：白腐真菌 + 漆酶　； 参考：《佳木斯大学》2017年硕士论文

【摘要】：随着农药大面积的使用,如何降低残留在环境中的农药成为现阶段重要科学研究内容,在多学科的研究中,生物降解无污染,广泛性以及高效性成为主要技术之一。白腐真菌作为一种生物降解菌,其通过产生的各种酶系,主要是漆酶,对农药有着高效的降解效果。因此,如何确定产漆酶白腐真菌及提高其产漆酶量成为研究降解农药的关键。本实验研究了在佳木斯四丰山采集的白腐真菌,应用愈创木酚培养基初步筛选出产漆酶的8种白腐真菌,进行培养基的优化,提高产漆酶量,应用ABTS法测定漆酶活性。并且将筛选出产漆酶最多的4个菌种对东北温室大棚土壤中农药残留嘧菌酯的降解,应用HPLC进行检测嘧菌酯残留量计算降解率,结果表明:1、将采集的白腐真菌分离纯化后接种到含愈创木酚的培养基上,能使培养基产生红色或淡紫色的有东方栓菌、多毛栓菌、桑黄、轮纹韧革菌、树舌、松生拟层孔菌、野生香菇、朱红密孔菌,这8种白腐真菌都产漆酶。2、根据培养基营养因素和培养条件的筛选得出:葡萄糖和硝酸钾是多数白腐真菌的碳氮源,金属阳离子Cu2+、K+、Mg2+对白腐真菌产漆酶有促进作用,而Ca2+对产酶有抑制作用,Zn2+对产漆酶没有影响。温度和pH对白腐真菌产漆酶有显著影响,在酸性条下,温度为25-30℃产漆酶量最佳。3、8种白腐真菌经过培养基优化后得出:东方栓菌产漆酶活性最高为133.14U/ml,是PDA培养基产漆酶活性的1.93倍;多毛栓菌产漆酶活性最高为73.05 U/ml,是PDA培养基产漆酶活性的1.50倍;桑黄产漆酶活性最高为31.48 U/ml,是PDA培养基产漆酶活性的1.85倍;轮纹韧革菌产漆酶活性最高为157.36 U/ml,是PDA培养基产漆酶活性的2.72倍;树舌产漆酶活性最高为59.05 U/ml,是PDA培养基产漆酶活性的1.52倍;松生拟层孔菌产漆酶活性最高为127.17 U/ml,是PDA培养基产漆酶活性的1.70倍;野生香菇产漆酶活性最高为448.16 U/ml,是PDA培养基产漆酶活性的1.42倍;朱红密孔菌产漆酶活性为137.77 U/ml,是PDA培养基产漆酶活性的2.36倍。4、将筛选出产漆酶最多的野生香菇、轮纹韧革菌、朱红密孔菌、东方栓菌用最佳培养基进行培养,提取粗酶液,对采集自温室大棚土壤中含残留嘧菌酯农药进行降解。结果表明:野生香菇的降解率最高,降解率达到了99.99%,轮纹韧革菌和朱红密孔菌的降解率都达到了90%以上,分别为94.35%和93.93%,东方栓菌的降解率最低为82.63%。
[Abstract]:With the extensive use of pesticides, how to reduce the residue of pesticides in the environment has become an important scientific research content at this stage. In the multidisciplinary research, biodegradation without pollution, extensiveness and efficiency has become one of the main technologies. White rot fungi, as a biodegradable fungus, produce various enzyme systems, mainly laccase, which have high degradation effect on pesticide. Therefore, how to determine laccase-producing white rot fungi and increase their laccase production is the key to study the degradation of pesticides. The white rot fungi collected in Sifeng Mountain of Jiamusi were studied in this paper. Eight kinds of white rot fungi producing laccase were preliminarily screened by guaiacol medium. The laccase production was improved by optimizing the culture medium, and the laccase activity was determined by ABTS method. Four strains producing laccase were selected to degrade the pesticide residue in greenhouse soil in Northeast China, and the degradation rate was calculated by using HPLC to detect the residue of pyridamole. The results showed that when the collected white rot fungi were isolated and purified and inoculated on the medium containing guaiacol, they could produce red or lavender oriental suppository, polychaete, mulberry yellow, rotund leatherback, tree tongue and pine laminopore. Wild Lentinus edodes and Phyllostoma sinensis, all of which produce laccase. 2. According to the nutrient factors of culture medium and the screening of culture conditions, glucose and potassium nitrate are the carbon and nitrogen sources of most white rot fungi. Metal cationic Cu2 + K + Mg2 promoted laccase production by white rot fungi, while Ca2 inhibited laccase production. Zn2 had no effect on laccase production. The effects of temperature and pH on laccase production of white rot fungi were significant. The laccase production of 8 species of white rot fungi was optimized at 25 鈩，

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