【作者】 陈平；

【导师】 黄乾明；

【作者基本信息】 四川农业大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 本文首先通过对粗毛栓菌形成菌丝球的物化条件、及不同物化培养条件下形成的菌丝球对Pb²⁺、Cu²⁺的吸附研究确定合适条件制备菌丝球;然后将菌丝球与培养好的蜡样芽孢杆菌共固定,制备杂合性生物吸附剂共固定菌。再以菌丝球、蜡样芽孢杆菌和共固定菌为研究对象,测定不同的吸附时间、pH、吸附剂浓度和金属离子浓度对三种生物吸附剂吸附Pb²⁺、Cu²⁺的影响,并对相关实验数据进行了Langmuir、Frendlich、Temkin和Henry吸附方程拟合。最后将三种生物吸附剂吸附Pb²⁺、Cu²⁺前后的红外吸收光谱进行了分析比较。取得以下主要研究结果:1.确定了0.3%硫酸铵、1%葡萄糖、0.1%吐温80和培养液pH4.5,在此基础上添加大量元素溶液、微量元素溶液和VB₁作为菌丝球培养液;在孢子接种浓度为10⁶个/mL,摇床转速150r/min的条件下培养96h,收集菌丝球,其细胞干重达到2.314g/L,菌丝球光滑、较硬、大小均匀、平均直径1.78mm;在菌丝球浓度2g/L（干重）,金属离子浓度50mg/L,pH 5.0的条件下吸附,用原子吸收分光光度法测定吸附体系总残留金属离子浓度,经计算,菌丝球对Pb²⁺、Cu²⁺的吸附量分别为17.93、10.87mg/g。2.菌丝球、蜡样芽孢杆菌和共固定菌分别吸附Pb²⁺、Cu²⁺时,发现在吸附剂浓度为2g/L（干重）,pH为5.0,金属离子浓度为50mg/L条件下吸附Pb²⁺1h(Cu²⁺2h),吸附效果良好,其吸附量分别为17.93、22.76、24.22mg/g和10.87、9.98、12.20mg/g。3.将菌丝球、蜡样芽孢杆菌和共固定菌分别对Pb²⁺、Cu²⁺的吸附数据进行Langmuir、Frendlich、Temkin和Henry方程拟合,除蜡样芽孢杆菌吸附Pb²⁺的Henry等温吸附线拟合相关系数r=0.776外,其余方程的拟合相关系数r都接近0.9或在0.9以上,四个方程都比较适合描述三种吸附剂对Pb²⁺、Cu²⁺的吸附过程。在三种吸附剂对Pb²⁺、Cu²⁺吸附的Langmuir方程拟合中,各方程特征常数K分别为:0.065、0.390、0.480和0.024、0.057、0.025L/mg;Q₀分别为:37.17、31.63、52.22和27.24、16.58、31.73mg/g。Pb²⁺与三种吸附剂结合的稳定性大小顺序为:共固定菌＞蜡样芽孢杆菌＞菌丝球。Cu²⁺与三种吸附剂结合的稳定性大小顺序为:蜡样芽孢杆菌＞共固定菌＞菌丝球。三种吸附剂对Pb²⁺的最大吸附量（mg/g）大小顺序为:共固定菌＞菌丝球＞蜡样芽孢杆菌。三种吸附剂对Cu²⁺的最大吸附量（mg/g）大小顺序为:共固定菌＞菌丝球＞蜡样芽孢杆菌。4.生物吸附剂菌丝球、蜡样芽孢杆菌和共固定菌的红外光谱图主要由蛋白质的吸收带、碳水化合物的吸收带和含硫、磷酸基团的吸收带组成。重金属离子除了与菌体之间产生静电吸引外,还与生物吸附剂的有机高分子物质及活性基团结合。各吸附剂吸附Pb²⁺、Cu²⁺时,起主要作用的官能团是羟基、羧基、磷酸基和含硫基团;蛋白质的氨基和其它分子的叔胺或磺胺基团更倾向于结合Cu²⁺。更多还原

【Abstract】 In this paper,In order to get the best physical and chemical conditions for the formation of T.gallica mycelium pellets and such pellets have a good effect for the Pb²⁺,Cu²⁺ adsorption,firstly we do some experiments by changing of the percentage of sulfate ammonia,glucose,Tween 80,culture medium pH,shaking bed rotating speed and spore inoculation concentration to check the results.Secondly take the mycelium pellets and Bacillus cereus together to culture one day to produce the hybrid biosorbent co-fixed microbe,the two combine efficiently to each other.For the Pb²⁺,Cu²⁺ adsorption of mycelium pellets,Bacillus cereus and the co-fixed microbe,we do some study on how do the different adsorption conditions like contact time,pH,biosorbent concentration or metal ion concentration affect Pb²⁺,Cu²⁺ adsorption.Experimental data related to the adsorption were used to fit the Langmuir,Frendlich,Temkin and Henry adsorption equation.Finally, the three biosorbents samples which were before and after Pb²⁺,Cu²⁺ adsorption were used to do the infrared adsorption spectra analysis.The results were as follows:1.The mycelium pellets culture medium with 0.3%sulfate ammonia,1%glucose,0.1% Tween 80 and pH4.5,on this basis,adding many elements solution,trace elements VB₁ as a solution and spore inoculation concentration 10⁶/mL,a shaking bed rotating speed 150 r/min under the conditions of 96 hours culturing,collected the mycelium pellets,the dry cell weight reached 2.314 g/L,the mycelium pellets were smooth,hard and of uniform size,with an average diameter of 1.78 mm;with 2g/L（dry weight）mycelium pellets.With 50 mg/L metal ion concentration and pH5,do adsorption under this condition,the residual metal ions concentration of the total biosorption system are determined by AAS,the uptake of the mycelium pellets’ adsorption to Pb²⁺,Cu²⁺ are respectively 17.93,10.87 mg/g.2.The three biosorbents（T.gallica mycelium pellets,Bacillus cereus,hybrid biosorbent co-fixed microbe,） were used to adsorb Pb²⁺,Cu²⁺ under different conditions. Through the study of contact time,pH,metal ions concentration and biosorbents concentration change to affect the biosorption,and found that the adsorption uptake increase with the contact time,pH and metal ions concentration increase,but decrease with the biosorbents concentration increase.It takes about one hour to reach balance for the three biosorbents to adsorb Pb²⁺,and about two hours for Cu²⁺.Under the condition of biosorbents’ concentration 2 g/L（dry weight）,pH 5.0,metal ion concentration 50 mg/L and contact time pb²⁺（one hour） Cu²⁺（two hour）,they adsorbed well,their adsorption uptake were 17.93,22.76,24.22 mg/g and 10.87,9.98,12.20 mg/g respectively.3.Use the data of the three biosorbents’ biosorption to Pb2^,Cu2 to fit Langmuir, Frendlich,Temkin and Henry equation,they fitted well,except the Henry fitting of Bacillus cereus’s biosorption to Pb²⁺ with the correlation coefficient value r=0.776.The rest of the correlation coefficient r of other equations fitting are close to 0.9 or more than 0.9.The four equations are suited to describe the three biosorbents’ adsorbion process for pb²⁺and Cu²⁺.In the Langrnuir equation fitting of the three biosorbents’ adsorption data,the equation feature constants K are:0.065,0.390,0.480 and 0.024,0.057,0.025L/mg;Q₀ are:37.17,31.63, 52.22 and 27.24,16.58,31.73 mg/g.The stability order of the three biosorbents’ Pb²⁺ adsorbtion is:co-fixed microbe＞Bacillus cereus＞mycelium pellets.While for Cu²⁺,the stability order is:Bacillus cereus＞co-fixed microbe＞mycelium pellets.The order of the max adsorption capacity of the three biosorbents to Pb²⁺ is:co-fixed microbe＞mycelium pellets＞Bacillus cereus.While for Cu²⁺,the order is:co-fixed microbe＞mycelium pellets＞Bacillus cereus.4.After the infrared spectra analysis and comparison of the data of mycelium pellets, Bacillus cereus and co-fixed microbe absorbed Pb²⁺ or Cu²⁺,and find that the infrared spectra of the biosorbents mainly contain protein adsorption band,carbohydrate components adsorption band,sulphur adsorption band and phosphate groups adsorption band.In addition to the static electronic attraction between the biosorbents and the ions,the ions still adsorbed to the active groups of the biosorbents.When the biosorbents adsorb Pb²⁺ and Cu²⁺,the functional groups such as hydroxyl,carboxyl,phosphoric acid and sulphur-based groups play an important role in the adsorption;while the amino group of protein and some other molecules’ amino-Amine or sulfa group are more inclined to combine Cu²⁺.更多还原