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假单胞菌WBC-3甲基对硫磷水解酶性质的初步研究 被引量:32
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作者 楚晓娜 张先恩 +2 位作者 陈亚丽 刘虹 宋冬林 《微生物学报》 CAS CSCD 北大核心 2003年第4期453-459,共7页
从最近分离到的有机磷农药降解菌Pseudomonassp .WBC 3中获得了甲基对硫磷水解酶 (Methylparathionhydrolase,MPH ,EC 3 1 8 3)。该酶在 48h的培养物中分布比例分别为 :上清液 2 1 % ,胞内 86 2 %和胞间质 1 1 7% ,说明MPH为胞内酶... 从最近分离到的有机磷农药降解菌Pseudomonassp .WBC 3中获得了甲基对硫磷水解酶 (Methylparathionhydrolase,MPH ,EC 3 1 8 3)。该酶在 48h的培养物中分布比例分别为 :上清液 2 1 % ,胞内 86 2 %和胞间质 1 1 7% ,说明MPH为胞内酶。经过CM sepharoseFastFlow阳离子交换层析 ,获得电泳纯的酶。SDS PAGE和凝胶过滤层析表明 ,该酶为单体蛋白 ,分子量约为 34kD。动力学分析显示该酶为非特异性有机磷降解酶 ,但最适底物为甲基对硫磷。在pH9~ 1 2范围 ,酶表现较高活力水平 ,最高活力的反应温度为 40℃。根据各类金属离子和鳌合剂对酶活的影响 ,推测MPH为金属酶。 展开更多
关键词 假单胞菌 磷水解酶 有机降解酶 有机农药 纯化 性质 微生物 降解
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参与碳氮磷转化的水解酶对不同施肥响应的差异 被引量:14
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作者 闵凯凯 何向阳 +4 位作者 吴倩怡 张冠友 胡锋 李辉信 焦加国 《土壤》 CAS CSCD 北大核心 2020年第4期718-727,共10页
本文旨在研究土壤水解酶对不同施肥的响应差异以及影响因素。通过在红壤中添加牛粪有机肥、化肥进行90 d的室内土壤培养试验,采用微孔板荧光法动态分析5、30和90 d参与碳氮磷转化的土壤水解酶(α-1,4-葡萄糖苷酶、β-1,4-葡萄糖苷酶、... 本文旨在研究土壤水解酶对不同施肥的响应差异以及影响因素。通过在红壤中添加牛粪有机肥、化肥进行90 d的室内土壤培养试验,采用微孔板荧光法动态分析5、30和90 d参与碳氮磷转化的土壤水解酶(α-1,4-葡萄糖苷酶、β-1,4-葡萄糖苷酶、纤维素酶、木聚糖酶、亮氨酸氨基肽酶、β-1,4-N-乙酰氨基葡萄糖苷酶、磷酸酶)活性。与不施肥(对照)相比,在30 d后,化肥处理的总酶活性显著下降,对应的参与碳氮磷转化酶活性均有不同程度下降;而有机肥处理的总酶活性在培养期内均未发生显著变化,但是其α-1,4-葡萄糖苷酶显著增加,而磷酸酶活性显著降低。参与碳转化的4种水解酶中,只有α-1,4-葡萄糖苷酶活性对施肥的响应较强,且施加有机肥增加其活性而无机肥则降低其活性;对于参与氮转化的水解酶而言,化肥明显抑制了亮氨酸氨基肽酶活性,而有机肥增加了β-1,4-N-乙酰氨基葡萄糖苷酶活性;磷酸酶活性明显受到有机肥的抑制作用,而对化肥的响应总体不明显。不同水解酶对不同施肥的响应有明显差异,NMDS分析表明,α-1,4-葡萄糖苷酶和亮氨酸氨基肽酶响应最明显,其次为磷酸酶与木聚糖酶;相关和冗余分析显示,土壤pH、可溶性有机碳对酶活性的影响最大,一定程度说明了不同肥料通过影响土壤理化性质进而影响水解酶活性。 展开更多
关键词 有机肥 化肥 碳氮转化水解酶 微孔板荧光法
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种植不同油茶品系实生苗对红壤胞外酶活性的影响 被引量:2
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作者 狄岚 王小东 +2 位作者 周莉荫 颜聪 肖相元 《经济林研究》 北大核心 2021年第2期74-81,共8页
【目的】为了解野生油茶与栽培种油茶对土壤物理、化学、生物性质的影响的差异情况,给栽培种油茶的选育提供一定的科学理论依据。【方法】以红壤土盆栽野生油茶和栽培种赣油1号和赣永6号油茶实生苗,研究了不同品系油茶种植土壤理化性质... 【目的】为了解野生油茶与栽培种油茶对土壤物理、化学、生物性质的影响的差异情况,给栽培种油茶的选育提供一定的科学理论依据。【方法】以红壤土盆栽野生油茶和栽培种赣油1号和赣永6号油茶实生苗,研究了不同品系油茶种植土壤理化性质和酶活性的差异情况。【结果】种植不同油茶品系实生苗对土壤pH值均无影响。种植野生油茶的土壤其有机质、全磷、有效磷和速效钾等养分含量均显著低于种植赣油1号和赣永6号油茶的土壤各养分含量,而栽培种赣油1号和赣永6号油茶的种植土壤中这些养分含量间均无差异。种植赣油1号油茶的土壤铵态氮含量低于赣永6号种植土壤的,但其硝态氮含量却显著高于赣永6号种植土壤的。种植野生油茶的土壤纤维二糖水解酶活性显著高于赣油1号和赣永6号种植土壤的,其增幅分别为18.76%和19.34%,而其土壤中β-1,4-葡糖苷酶和β-1,4-木糖苷酶活性均显著低于赣油1号和赣永6号种植土壤的。种植赣永6号的土壤氮水解酶(β-乙酰葡糖胺糖苷酶和亮氨酸氨基肽酶)的活性显著高于野生油茶和赣油1号油茶种植土壤的,其增幅分别为9.45%、14.34%、8.57%和10.39%,而种植野生油茶和赣油1号油茶的土壤亮氨酸氨基肽酶和β-乙酰葡糖胺糖苷酶活性之间的差异均不明显。种植野生油茶的土壤磷酸单酯酶活性显著低于赣油1号和赣永6号油茶种植土壤的磷酸单酯酶活性,其降幅分别为20.88%和13.91%。【结论】栽培种赣永6号油茶对土壤地力的影响程度比野生油茶和栽培种赣油1号油茶对土壤地力的影响程度大。 展开更多
关键词 油茶 水解酶 水解酶 磷水解酶
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Lipase and Phospholipase in the Hydrolysis of Lipids in Wastewater from Swine Slaughterhouse and Subsequent Biological Treatment Study
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作者 Aniela Pinto Kempka Rafael Celuppi Jaime Humberto Palacio Revello 《Journal of Agricultural Science and Technology(A)》 2013年第10期757-765,共9页
Wastewaters from slaughterhouses are characterized by a high concentration of oils and greases that can cause problems in conventional treatments. The degradation of difficult pollutants in more easily degradable prod... Wastewaters from slaughterhouses are characterized by a high concentration of oils and greases that can cause problems in conventional treatments. The degradation of difficult pollutants in more easily degradable products can be mediated by enzymes. Enzymatic hydrolysis facilitates the biodegradation because it makes the organic material is made available as compounds of lower molecular weight, the lipids being made available in the form of free fatty acids. The application of enzymes as processing aids in wastewater treatment has some advantages, such as the specificity that allows control of the products, which leads to increased income generation for the non-toxic byproducts and moderate conditions of operation. The objective of this work was optimizing the conditions for lipids enzymatic hydrolysis present in swine slaughterhouse wastewaters using lipolytic enzymes and subsequent biological treatment study. Temperature, pH and enzyme concentration were the variables tested. The enzymes showed good activity and could therefore be used for the hydrolysis process proposed here. The optimized conditions to maximize the release of fatty acids were: temperature of 36 ℃, pH 8.5 and enzyme concentration of 1.1%, yielding fatty acids in the order of 31.50μmol/mL for lipase and 31.13 μmol/mL for phospholipase. All variables influenced the release of fatty acids. The maximum yield of biogas was 89.65 mL in the reactor added the sludge, raw wastewater and phospholipase in the conditions optimized the hydrolysis step, obtaining a chemical oxygen demand (COD) reduction of 90.01% in relation to the value of the COD of the raw wastewater. 展开更多
关键词 WASTEWATER HYDROLYSIS LIPIDS LIPASE PHOSPHOLIPASE biological treatment.
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Adeno-associated virus vector-mediated triple gene transfer of dopamine synthetic enzymes
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作者 樊东升 沈扬 康德宣 《Chinese Medical Journal》 SCIE CAS CSCD 2001年第12期44-47,105,共5页
Objective To explore triple gene transfer of dopamine synthetic enzymes with separate adeno-associated virus (AAV) vectors.Methods The genes for dopamine synthetic enzymes, tyrosine hydroxylase (TH), aromatic L-amino ... Objective To explore triple gene transfer of dopamine synthetic enzymes with separate adeno-associated virus (AAV) vectors.Methods The genes for dopamine synthetic enzymes, tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), and GTP cyclohydrolase Ⅰ (GCH, an enzyme critical for tetrahydrobiopterin synthesis) were cotransduced into 293 cells with separate AAV vectors. Expressions of TH, AADC and GCH were detected by Western blot analysis. Intracellular dopamine level was assayed by high-performance liquid chromatography.Results TH, AADC and GCH were effectively coexpressed in transduced cells with three separate AAV vectors, AAV-TH, AAV-AADC and AAV-GCH. Furthermore, the coexpression resulted in an effectively spontaneous dopamine production in cotransduced cells.Conclusion The triple transduction of TH, AADC and GCH genes with separate AAV vectors is effective, which might be important to gene therapy for Parkinson's disease. 展开更多
关键词 dopamine · GTP cyclohydrolase · adeno-associated virus
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Transient expression of organophosphorus hydrolase to enhance the degrading activity of tomato fruit on coumaphos 被引量:2
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作者 Jie-hong ZHAO De-gang ZHAO 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2009年第2期142-146,共5页
We constructed an expression cassette of the organophosphorus pesticide degrading (opal) gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expr... We constructed an expression cassette of the organophosphorus pesticide degrading (opal) gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expression system. β-Glucuronidase (GUS) staining, reverse transcription-polymerase chain reaction (RT-PCR), wavelength scanning, and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on coumaphos of organo- phosphorus hydrolase (OPH) in tomato fruit. The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 Uhng total soluble protein. These results will allow us to focus on breeding transgenic plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields. 展开更多
关键词 BIOREMEDIATION E8 promoter Organophosphorus hydrolase (OPH) Transient expression
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