A spectrophotometric method for detection of enzymatic degradation of thin polymer films

An assay method has been developed for monitoring the enzymatic degradation of thin films of translucent polymers. The method was based on the observation that when a solution-cast film of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was exposed to a solution of a depolymerase fromPseudomonas lemoignei, the surface of the film roughened and the film became visibly turbid. This increase in turbidity could be measured spectrophotometrically and was reproducible during the initial stage of degradation. Turbidity correlated very closely with film weight loss early in the degradation but reached a maximum value before extensive degradation had taken place. For a given set of films, this correlation was independent of the concentration of the enzyme used, although it did vary with the mode of enzyme exposure. The turbidity was associated with the exposure of crystalline domains due to the removal of amorphous material from the film surface. The increase in crystallinity at the surface was verified by attenuated total reflectance infrared spectroscopy (ATRIR). In conjunction with SEM, weight loss, and ATRIR, the film turbidity assay provided much semiquantitative insight into the mechanism of the enzymatic degradation reaction. This assay was used to study the enzymatic degradation of films of PHBV solution blended with cellulose acetate esters (CAE). The presence of only 25% of CAE of degree of substitution 2.9 severely hampered the enzymatic degradability of PHBV, a result which is consistent with the environmental degradation of these same samples exposed to activated sludge.     

Physical properties and biodegradability of blends containing poly(ε-caprolactone) and tropical starches

In order to assess feasibility of tropical starches (sago and cassava starches) as biodegradable plastic materials, blending with poly(-caprolactone) (PCL), a biodegradable polymer, was carried out. It was confirmed that the physical properties (tensile strength and elongation) of PCL/sago and PCL/cassava blends were similar to those of PCL/corn blend, suggesting that sago and cassava starches can also be blended with PCL for production of biodegradable plastic. However, the properties of all PCL/starch blends were still low compared with those of polyethylene. Enzymatic degradability evaluation showed that lipase degradation of PCL and-amylase degradation of starch increased as the starch content in the blend increased. Burial test of the blends for 1, 3, and 5 months was carried out and the rate of degradation of the PCL/sago blend was confirmed to be slower than those of PCL/corn and PCL/cassava blends. Observation of the film blends structure by scanning electron microscope revealed that the starch was dispersed in a PCL continuous phase. Furthermore, changes in the film surface before and after enyzme treatments were observed.     

Cd胁迫下城郊农业土壤微生物活性对生物炭输入的响应

为了探究Cd胁迫下生物炭输入对城郊农业土壤微生物活性的影响,通过室内培养试验,分析CK处理(风干土壤)、Cd处理(风干土壤+外源Cd)、CdBC处理〔风干土壤+外源Cd+2%生物炭,即生物炭与土壤(以湿质量计)质量比为2%〕、BC处理(风干土壤+2%生物炭)对土壤呼吸、土壤微生物数量及土壤酶活性的影响. 结果表明:整个培养期(0~60 d)内,各处理下土壤CO₂累积释放量表现为CK处理＜Cd处理＜CdBC处理＜BC处理. 与CK处理相比,CdBC处理下生物炭输入能显著增加土壤CO₂累积释放量(P＜0.05),增幅为43.6%,并且土壤中细菌、放线菌、真菌数量也有显著增加(P＜0.05),增幅分别达到12.7%、62.7%、18.7%. Cd胁迫对土壤酶活性的抑制表现为蔗糖酶＜脲酶＜中性磷酸酶,抑制率分别为3.5%、6.8%、18.0%. 生物炭输入可使受Cd胁迫的土壤脲酶、蔗糖酶的活性有所增强,增幅分别为15.0%、18.4%. 可见,生物炭输入可在不同程度上缓解Cd胁迫对蔗糖酶、脲酶活性及土壤微生物数量的影响.      

正交实验选择纤维素酶产生菌的最优综合培养条件

在复合碳源、30℃恒温培养条件下,用正交实验法对影响纤维素酶产生菌降解纤维素的5种单因素培养条件进行了优化,并将优化得到的条件应用于厌氧-好氧废水处理系统.结果表明,MgSO4用量等单因素对纤维素酶活性以及纤维素降解率有不同程度的促进作用;正交实验得到厌氧菌和好氧菌的最佳培养条件不完全一致;在废水处理系统中,5种单因素的最佳综合水平为:30mg/L MgSO4,20mg/L CoCl2,CNP配比为400:5:1,氮源为NH4Cl (28.7mg/L), pH=7.0,此时厌氧菌及好氧菌酶活性分别为4801U/L和4794U/L,酶稳定性分别达到91.0%和95.5%,纤维素降解率为31.9%和28.4% .     

一种嗜热细菌来源角质酶的分离纯化及酶学性质

通过跟踪发酵液中pNPB水解酶活性,对角质诱导的Thermobifida fusca 口发酵液进行分离纯化.采用活性炭脱色、硫铵沉淀、Phenyl HP疏水色谱、DEAE sephamse阴离子交换色谱等方法,分离纯化得到电泳纯PNPB水解酶.该酶水解角质可得到角质单体,是一种角质酶.SDS-PAGE电泳结果显示,角质酶表观分子量约为29×10~3.该酶的最适温度为60℃.在40℃和60℃下均具有良好的热稳定性.最适pH为8.0,pH稳定范围为6.0～9.0.该角质酶的生化性质适合在纺织工业中应用.图8表2参17     

黄土丘陵区撂荒农田土壤酶活性及酶化学计量变化特征

明确土壤胞外酶活性及酶化学计量比在农田撂荒过程中的变化特征及驱动因素,对揭示土壤养分可利用性随植被恢复的变化规律和阐明生态系统养分循环机制有重要意义.本文以黄土丘陵区不同撂荒年限坡耕地（0、10、20和30 a）为研究对象,通过测定土壤酶活性及酶计量比、土壤理化性质、植物多样性和科组成,探讨农田撂荒后土壤微生物养分限制状况及酶活性和酶计量比的主控因子.结果表明,β-1,4-N-乙酰氨基葡萄糖苷酶（NAG）、亮氨酸氨基肽酶（LAP）和碱性磷酸酶（ALP）的活性均随撂荒年限增加而显著增加,而β-1,4-葡萄糖苷酶（BG）活性则相反.土壤BG：（NAG+LAP）和BG：ALP呈现出与土壤BG活性相同的随撂荒年限增加而减小的趋势;而（NAG+LAP）：ALP呈先升后降趋势,在撂荒20 a时达到最高值.酶化学计量的向量长度随撂荒年限增加而显著减小,表明撂荒过程中土壤微生物受碳（C）限制程度的减弱.撂荒前10 a酶化学计量的向量角度小于45°,后20 a大于45°,说明撂荒前期微生物受氮（N）限制,后期受磷（P）限制.冗余分析结果揭示土壤有机碳含量、总氮含量、C：N、C：P、pH以及植被多样性是调控酶活性和酶计量比变化的主要因子.分类变异分析表明,土壤和植物因素可解释不同撂荒年限下土壤酶活性及酶计量比差异的62.0%,植被特征与土壤理化特性之间的交互作用是土壤酶活性及酶计量比变化的主控因子,解释度为37.1%.综上所述,黄土丘陵区农田撂荒过程中应考虑外源P的投入以缓解生态系统中有效P的不足,研究结果可为理解脆弱生境恢复生态系统中微生物介导的生物地球化学循环机制以及指导土壤养分管理和生态可持续发展提供理论依据.     

河流湿地土壤芘的降解和生物有效性模型与酶活性动态变化(Model of Pyrene Degradation and Bioavailability and Dynamic Changes of Enzymatic Activities in the Riverine Wetland Soil)

在淹水和非淹水条件下,在河流湿地土壤中加入高浓度(10mg·kg-1)和极高浓度(200mg·kg-1)芘,研究了芘降解过程、芘生物有效性和土壤酶活性的长期动态变化.结果表明,培养91d后,高浓度芘处理的土壤中芘的残留率在淹水和非淹水条件下分别为79.0%和51.4%,而极高浓度处理的芘残留率在淹水和非淹水条件下分别为61.1%和33.9%.利用Tenax-ta树脂提取快速解吸态生物有效芘,结果发现,有效芘浓度在高浓度芘处理的两种土壤中的值比较接近,分别为1.01mg·kg-1和1.23mg·kg-1;而有效芘浓度在极高浓度处理的两种土壤的相应值差别较大,分别为20.31mg·kg-1和9.44mg·kg-1.采用四参数Logistic模型能够精确地拟合芘降解过程和生物有效性变化过程(p<0.0001).土壤蔗糖酶、脲酶和多酚氧化酶对芘污染敏感,而过氧化氢酶对芘污染不敏感.实验结果揭示了河流湿地土壤芘在淹水条件下降解活跃阶段持续时间短和芘生物有效性高的特征以及土壤蔗糖酶、脲酶和多酚氧化酶活性可以作为评估芘污染影响的候选指标.     

木质纤维素生产燃料乙醇的关键技术研究现状

木质纤维素是自然界广泛存在且廉价的可再生资源.其主要成分纤维索、半纤维素是潜在的燃料乙醇生产原料.虽然由木质纤维素生产燃料乙醇的技术路线已具可行性,但存在着具体工艺环节复杂、生产能耗高等局限,严重阻碍了其规模化生产.目前纤维素燃料乙醇生产主要围绕预处理、酶解、发酵三大关键步骤进行技术攻关,其中预处理的能耗和效率、发酵过程的五碳糖利用等问题成为该工艺的重要制约因素.本文在综述国内外纤维素乙醇生产关键步骤的基础上,着重分析了各种物理、化学和生物预处理的优缺点以及新兴的预处理思路,归纳了各类纤维素乙醇生产菌的特点,包括耐高温和五碳糖的利用,并介绍了当前主要的发酵方式和优化措施,以期为木质纤维素生产乙醇提供新的研究思路.表5参86     

甲磺隆结合残留对土壤酶活性的影响

为了研究甲磺隆结合残留对土壤酶活性的影响,将黄粉土(水稻土)经甲磺隆结合残留处理98天,检测土壤脲酶、酸性磷酸酶、过氧化氢酶和蔗糖酶活性.结果表明,甲磺隆结合残留对土壤酶活性的影响不仅与甲磺隆结合残留处理浓度和培养时间有关,同时还与土壤酶种类有关.培养前期(第14天前),甲磺隆结合残留对土壤酸性磷酸酶和过氧化氢酶具有抑制作用,而对土壤脲酶和蔗糖酶的影响则表现为低浓度时激活,高浓度时抑制;培养后期(第14天后),甲磺隆结合残留对土壤脲酶、酸性磷酸酶和过氧化氢酶具有激活作用,在一定浓度范围内(0.050~0.281mg·kg-1)对土壤蔗糖酶也具有一定激活作用;就土壤酶而言,在污染早期,土壤过氧化氢酶对甲磺隆结合残留较敏感,可以作为表征甲磺隆结合残留土壤污染的生物活性指标.     

Enzymatic treatment of estrogens and estrogen glucuronide

Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers.In this article we investigated the enzymatic treatment of three estrogens (estrone,17β-estradiol,and 17α-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen.The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay.In addition,we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-(β-D-glucuronide) degradation.The two enzymes eliminated 17β-estradiol 3-(β-D-glucuronide) and the intermediate,17β-estradiol,efficiently.