白腐真菌降解染料反应器微生物群落结构研究

利用DGGE分子标记技术分析了两种白腐真菌生物反应器处理染料废水过程中生物膜微生物群落结构的动态变化及其差异性,结果表明:运行方式和染料废水浓度对敞开式白腐真菌生物反应器处理染料废水过程中生物膜微生物群落结构及其变化趋势有一定影响。序批式反应器运行的起始阶段其生物膜上黄孢原毛平革菌在数量上占绝对优势,但约1个月后逐渐侵染了热带假丝酵母、白地霉和哈茨木霉等杂菌;连续运行的白腐真菌生物反应器虽然在起始阶段存在哈茨木霉等杂菌的侵染,但约3个月后杂菌被逐渐淘汰,黄孢原毛平革菌在数量上重新占据优势,使其有利于发挥高效的降解功能。     

Biosorption of lead by Phanerochaete chrysosporium in the form of pellets

The growth of Phanerochaete chrysosporium(ATCC 24725) in pellets was influenced by culture time,medium pH,C/N,surfactant concentration,spore number in inoculum,and shaking rate.The removal of Pb^2 from aqueous solution by this kind of mycelial pellets was studied.The results indicated that many factors affected biosorption.These factors included pH,Pb^2 concentration,co-ion,adsorption time, and chemical pretreatments of biomass.Under optimum biosorption conditions(pH4.5,27℃,16h),the highest lead uptake of 108 mg/g,was observed with mycelial pellets of 1.5-1.7 mm in diameter which were treated with 0.1 mol/L NaOH solution before adsorption.Pretreatment of biomass with NaOH further increased its biosorption capacity.     

黄孢原毛平革菌酵母双杂交cDNA文库的构建及应用

作为研究木质素降解系统的模式微生物黄孢原毛平革菌,迄今还没有蛋白-蛋白相互作用方面的报道.本研究利用酵母双杂交技术构建了低氮培养基中培养2d和3d的酵母双杂交cDNA文库,分别得到2.5×105和3.0×105个重组子.以14-3-3蛋白为钓饵筛选3d cDNA文库,在SD/-Ade/-His/-Leu/-Trp缺陷培养基平板上共获得了约600个单克隆,进一步分析发现,有30个克隆可激活3个报告基因.分离自这些克隆中的质粒DNA能转化大肠杆菌.利用HaeⅢ和Sau3AⅠ限制酶切分析可将它们分为4类.序列测定和同源分析结果表明,其中一类为14-3-3蛋白基因,提示14-3-3蛋白可形成二聚体,另外3类在GenBank没有明显同源的基因.通过SBASE网站预测,编号为Y2H333的克隆含有一个OB-fold核酸结合结构域(OB-fold nucleic acid binding domain).这些研究结果表明,所构建的黄孢原毛平革菌cDNA酵母杂交文库是成功的,14-3-3蛋白在黄孢原毛平革菌中能以蛋白-蛋白相互作用的形式发挥功能.图4表1参28     

黄孢原毛平革菌反复脱色模拟染料废水的试验研究

研究了白腐真菌黄孢原毛平革菌在35℃、150r／min条件下培养3d后，培养基类型、染料浓度以及反复脱色次数对其脱色的影响。玉米芯浸出液是首选的可替代营养源的物质，培养3d的黄孢原毛平革菌6h脱色率即达92％。黄孢原毛平革菌能够在短时间(6h)基本脱色较高浓度(40mg／L，60mg／L)的活性艳红K-2BP模拟染料废水，对于低浓度(10mg／L，20mg／L)脱色作用则比较缓慢，浓度超过150mg／L即显示出对菌的毒性作用。玉米芯浸出液培养3d的菌可以重复脱色模拟废水6次以上，每批脱色率均在82％以上，但菌球的沉降性能变差。     

黄孢原毛平革菌对蒽的降解研究

研究了不同培养条件下２种品系黄孢原毛平革菌对蒽的降解效果,研究结果表明,各样品中蒽的降解率为７５％～９９％;动培养方式比静培养方式对蒽的降解效果好;ＯＧＣ１０１菌优于ＢＤＭ－Ｆ－１７６７菌;对菌进行５０℃～９９％;动培养方式比静培养方式对蒽的降解效果好;ＯＧＣ１０１菌优于ＢＫＭ－Ｆ－１７６７菌;对菌进行５０℃、２４h的 预处理,降解系统的主要酶仍保持一定的活性和稳定性。     

白腐菌和红平红球菌共固定化处理增塑剂废水

以赭曲霉死菌丝体为载体,吸附红平红球菌(Rhodococcus erythropo lisNG0402)和白腐菌(Phanerochaete chrysosporiumM E-446)的混合物,用聚乙烯醇作包埋剂进行共固定化。用制备好的共固定化细胞处理增塑剂废水,连续运行19d的结果表明,共固定化处理增塑剂废水的效果远好于单一菌的效果。在pH7.0、溶解氧质量浓度3.0m g/L、水力停留时间25h的条件下,废水中苯酚、COD和色度的去除率连续15d达到了90%以上,对苯酚的去除能力为2.16kg/(m3.d)。     

非灭菌环境投加染料时间对白腐真菌降解活性染料的影响

应用氮限制液体培养基(C/N=56／8．7)研究了非灭菌环境投加染料时间对白腐真菌Phanerochaete chrysosporium降解活性艳红K-2BP染料的影响．试验结果表明,纯培养2d和3d后,在非灭菌环境投加活性艳红K-2BP染料的体系脱色率与在灭菌环境投加该染料的体系基本相当,其5d脱色率在90％以上;而纯培养ld时在非灭菌环境投加染料体系的脱色率却只有8l％．引起纯培养ld体系脱色率降低的主要原因是活性艳红K-2BP对白腐真菌Phanerochaete chrysosporium的生长有抑制作用和反应体系感染了酵母菌．但是,如果纯培养延长至2d以后,由于白腐真菌菌丝体已在体系内占优势,尽管在非灭菌环境下还有酵母菌侵入,但它不会占优势,从而也就不会影响白腐真菌对染料的降解作用．因此,在氮限制液体培养基中,只要白腐真菌Phanerochaete chrysosporium在反应体系占优势,就可以适当缩短纯培养时间,提前在非灭菌环境投加染料,使白腐真菌Phanerochaete chrysosporium的培养和对活性染料的脱色同步进行．     

Degradation kinetics of terephthalic acid wastewater with the hybrid strains constructed with Phanerochaete chrysosporium

The degradation kinetic parameters in terephthalic acid (TPA) wastewater for the hybrid strains of Fhh and Fhhh obtained through the protoplast fusion of the fungi Phanerochaete chrysosporium (PC) and Saccharomyces cerevisiae Y99, and the native bacteria YZ1 were measured in this research. The highest level of the specific degradation rate for Fhhh and Fhh during 20h reaction were 0.2238 and 0.2163 h^‐1, which were higher than that of their three parental strains and that of the anaerobic bacteria reported. It suggested that the abilities of growth and degradation for Fhh and Fhhh in TPA wastewater were better than their three parental strains. They could create potentials for the purification of TPA wastewater with higher efficiency.     

Optimal parameters for Phanerochaete chrysosporium degradation of terephthalic acid wastewater

The optimal parameters for the degradation of pure terephthalic acid (PTA) wastewater by the white rot fungus Phanerochaete chrysosporium were investigated. The relative mathematical model, objective function equation, constraint equations, constraint conditions and computer program were established in the optimal method. The results showed that the optimal value of the reactor volume needed was 9798 m³. It could save 83.82% of the reactor volume, which was needed for the native bacterium YZ1. This optimal method is simple, useful and economical for average treatment plants. Using the optimal parameters and P. chrysosporium to design and operate the treatment process of PTA wastewater would increase the treatment efficiency and decrease the cost.     

活性黑对黄孢原毛平革菌锰过氧化物酶的影响

观察了活性黑KN-B(Reactive Rlack KN-B,RB KN-B)对黄孢原毛平革菌(Phanerochaete chrysosporium)锰过氧化物酶(MnP)酶活力和菌丝超微结构的影响以及黄孢原毛平革菌对RB KN-B的降解.于P.chrysosporium培养液MnP酶活达最高前,分别加入质量浓度为50 mg/L,200 mg/L,350 mg/L和500 mg/L的RB KN-B.分光光度法检测培养液MnP酶活,电镜观察菌丝超微结构的影响,紫外-可见光谱法检测培养液中RB KN-B的降解.结果显示,1)与对照组相比,50 mg/L RBKN-B组的MnP酶活力增强,200 mg/L、350 mg/L和500 mg/L组的MnP酶活力均显著低于对照组;2)电镜观察显示,经RB KN-B作用后,菌丝细胞膜受损,细胞内含物减少,胞质浓缩,出现质壁分离现象,500mg/L组有大量细胞解体;3)紫外-可见光谱扫描显示,RB KN-B经黄孢原毛平革菌降解,可见光波段最大吸收峰由598 nm移至525 nm和556 nm,峰值减小,紫外波段的吸收峰由315 nm移至352 nm.结果显示,黄孢原毛平革菌对RB KN-B的反应类似机体对不良环境因子的应激反应,经历了诱导、抑制及衰退的过程;RB KN-B对黄孢原毛平革菌菌丝细胞超微结构的损伤随RB KN-B浓度增高而增强,表明RBKN-B对MnP酶活的抑制与黄孢原毛平革菌结构受损密切相关;黄孢原毛平革菌对RB KN-B具有一定的降解能力,其中MnP作为关键酶起了重要的作用.