木质素过氧化物酶和锰过氧化物酶对染料脱色的性能比较

通过控制培养基中Mn^2 的浓度获得只含有木质素过氧化物酶（LiP)或锰过氧化物酶（MnP)的单一酶培养物，用于比较LiP和MnP对染料降解脱色的条件和能力的差异，结果表明，LiP对橙Ⅰ降解脱色的最佳条件为H2O20.15mmol/L,pH3.0温度40℃，MnP为H2O20．15mmol/L,pH3.5，温度40℃，最佳条件下，考察LiP和MnP作用下染料浓度和染料类型对脱色的影响，发现LiP作用下脱色率明显高于MnP作用下的脱色率，表明LiP可作用的染料浓度比MnP高，可作用的染料范围比MnP大，由此认为LiP具有比MnP更强的脱色能力。     

木质素降解酶对四环素的降解可行性

选取了使用广泛、在环境中残留量大、生物毒性大、不易被生物降解的四环素类抗生素中的四环素,考察了其被木质素过氧化物酶(LiP)和锰过氧化物酶(MnP)粗酶液体外降解的效果,证明2种酶对四环素均有较强的降解作用,并在此基础上比较了3个不同酶活(15 U·L-1、40U·L-1、70 U·L-1)下的降解率.结果表明,同等酶活(40 U·L-1)下,LiP比MnP的降解能力强,在适宜的反应条件下,LiP对50 mg·L-1四环素的降解率在5min内可达95%以上,而MnP在2h后仅达70%.对于LiP,当酶活从15 U·L-1升高至40 U·L-1时,降解率提高约15%,当酶活进一步升高至70 U·L-1时,降解率变化不大,而提高酶活可明显提高MnP对四环素的降解率.此外,通过反应进程中补加H2O2,可以明显提高MnP对四环素的降解率.     

葡萄糖补料对白腐真菌P.chrysosporium产木质素降解酶的影响

为了提高木质素降解酶的产量,采用氮限制培养基(C/N=56.0/8.8),研究了葡萄糖补料对P.chrysosporium产木质素降解酶的影响.结果表明,在培养第1d即接种24h后进行葡萄糖补料,可以刺激菌丝体的生长和产酶,补料浓度达到2g·L-1时,补料体系中的MnP酶活可提高至空白对照样(不进行补料)的2.5倍.同时,对葡萄糖分批补料方式进行的试验结果显示,培养过程中每24h进行1次补料,葡萄糖补料终浓度为1.5g·L-1的补料方式,与每48h的补料方式相比,产酶效果更好.MnP酶活峰值和达到峰值时酶的总产量可提高至空白对照样(不进行补料)的2.7倍和3.0倍,且酶活能够在200U·L-1以上稳定4d.实验结果说明,葡萄糖补料可以有效刺激白腐真菌产酶,且连续低浓度葡萄糖补料可获得较优的产酶效果.     

金鱼藻过氧化物酶降解双酚A的特性

本文对金鱼藻（Ceratophyllum demersum）过氧化物酶（POD）进行了分离纯化及酶学特性分析,并探讨了其对双酚A （BPA）的降解特性.结果表明,采用硫酸铵和AKTA蛋白纯化系统获得纯化的POD,该酶具有较广的温度和pH值适应范围,在温度为70℃,pH值为7,H₂O₂浓度为5mmol/L时POD酶活力最大.金鱼藻POD可高效降解BPA.当金鱼藻POD活力为10U/mL时,15min内对0.02mmol/L BPA的降解率达到99.67%,活力为25U/mL及以上时,完全降解.降解条件的最适范围分别为H₂O₂ 1~5mmol/L、30~60℃、pH 5~7.其降解动力学米氏K_m为0.09mmol/L,最大反应速度为9.71mmol/（L·h）.BPA经金鱼藻POD降解后无雌激素效应.金鱼藻POD高稳定性和高活性的特点以及对BPA的高效降解能力是该水生植物成为水环境BPA污染的理想修复工具的重要原因.     

木质素过氧化物酶降解四环素机理的研究

为探究过氧化物酶对四环素类抗生素的降解机理,以白腐真菌Phanerochaete chrysosporium表达的木质素过氧化物酶(LiP)粗酶为研究对象,研究其产酶条件及其降解四环素(TC)的机理。结果表明:低浓度Mn2+可促进产酶,碳源、氮源限制是产酶的重要条件;降解反应进行10min后,H2O2消耗至阈值0.045mmol/L,LiP对TC的降解停止,其降解率达到82%;TC降解反应趋于平衡时,补加TC和H2O2后,LiP依然可以高效发挥作用,其二次降解率达到65%;LiP作用于不同初始浓度(10mg/L、25mg/L、50mg/L、100mg/L)TC,显示LiP对不同浓度TC均可有效降解,TC降解率分别为62%、80%、82%和90%;降解产物的抑菌性试验反映出降解产物较TC母体的抑菌性明显降低;LC/MS捕获到6种可能的降解产物E-TC、TP 461、E-TP 461、TP 477、TP 475和TP 445,并推测出LiP降解TC可能的降解途径。     

白腐真菌批式发酵动力学模型研究

为了进一步认识白腐真菌发酵产酶机理并指导相关发酵工艺设计,在一系列试验研究的基础上,建立了描述黄孢原毛平革菌批式发酵过程的动力学模型,并采用自由悬浮和固定化2种培养条件下的试验数据分别回归了模型各参数.模拟计算结果表明,生物量、葡萄糖、氨氮、木质素酶浓度的的计算值与实测值的偏差在15%以内.对比2种培养条件下的模型参数发现,二者得到的最大生物量浓度均为1.78 g/L;固定化培养菌体的比增长速率值(0.668 3 d-1)大于自由悬浮培养值(0.514 4d-1);自由悬浮培养中绝大部分葡萄糖消耗于非生长菌体代谢,固定化培养中则部分葡萄糖用于菌体生长,且氨氮消耗更快;菌体的产酶过程与生长无关;自由悬浮培养条件下,菌丝小球可合成MnP(231 U/L),其MnP合成能力为115.8 U·(g·d)-1,且高峰后依然可以保持26.1 U·(g·d)-1,适合采用补料-分批发酵的模式提高MnP的产量及其发酵效率;固定化培养条件下,菌体可同时合成MnP(410 U/L)和LiP(721 U/L),但其LiP和MnP合成能力在酶活高峰之后明显下降,分别由248.9 U·(g·d)-1和80.1 U·(g·d)-1下降到0 U·(g·d)-1和6.04 U·(g·d)-1,故而在考虑补料-分批发酵模式时,补料时机应选择在培养中期的LiP与MnP峰值之前.     

正交实验选择嗜碱细菌降解木质素的

本文在复合碳源的碱性液体培养条件下(pH≈10.5),用正交实验法对嗜碱木质素降解细菌6号菌株降解木质素的金属离子培养条件进行了优化.结果表明,各金属离子对6号菌株产生的木质素降解酶Lac-case和MnP以及木质素降解率都有一定促进作用.分析正交实验结果表明,其中Mn2+和Cu2+是量主要的影响因素.各种金属离子的最佳综合水平为MnSO4@H2O0.7g/L、CuSO4@5H2O1mg/L、FeSO4@7H2O5mg/L、ZnSO4@7H2O30mg/L、CaCl2@2H2O50mg/L.     

白腐菌锰过氧化物酶对2,2’,4,4’-四溴联苯醚的降解

多溴联苯醚(PBDEs)是一类在环境中普遍存在的全球性有机污染物,对这类污染物环境行为的研究虽然已有较多报道,但是微生物降解方面尤其是针对单一胞外酶降解的研究还比较少。文章以白腐菌模式菌种Phanerochaete chrysosporium为对象,研究其分泌的胞外酶锰过氧化物酶(MnP)对环境中最常检测到的2,2’,4,4’-四溴联苯醚(BDE47)的好氧降解,考察不同β-环糊精浓度对该酶降解BDE47的影响,并初步探讨了胞内细胞色素P450的作用。结果表明,MnP能有效降解BDE47,在培养15 d后,扣除空白损失,降解率达到70%左右。低浓度和高浓度的环糊精对MnP降解BDE47无显著性影响。胞内细胞色素P450对BDE47的降解没有明显贡献。     

黄孢原毛平革菌固态发酵产漆酶的研究及应用

对黄孢原毛平革菌（BKMF-1767）利用香蕉皮和玉米棒为发酵底物产胞外漆酶进行了研究.结果表明,当香蕉皮与玉米棒混合比例为1∶2、诱导剂CuSO₄为0.4 mmol/L时,能获得最高漆酶酶活12.68　U/g.利用固态发酵所获得的粗漆酶液,进行了降解五氯酚的试验.在没有氧化还原介体时粗漆酶液能降解PCP,粗酶液中加入5　mmol/L氧化还原介体（ABTS）能获得更高的降解率,反应6 h分别为37.8%和97%.将粗漆酶液用(NH₄)₂SO₄盐析纯化,用提纯后漆酶降解PCP,6 h后降解率为81.8%.     

裂褶菌F17对偶氮染料刚果红的脱色降解及其产物分析

利用本实验室新构建的染料脱色降解体系,对裂褶菌F17(Schizophyllum sp.F17)脱色降解刚果红进行了研究,分析了该菌的主要降解酶,并对刚果红降解产物进行分离和鉴定.结果表明,裂褶菌F17在此体系中对刚果红表现出较高的脱色降解能力,菌球加入48h后脱色率达到91.5%;酶活检测表明,该菌主要产生锰过氧化物酶(MnP),并在脱色48h时,MnP酶活达到最大值96.1U·L-1.此外,对脱色96h和192h后的脱色液进行紫外-可见扫描,发现刚果红在可见光区495nm处的吸收峰已消失,并在紫外区出现多个吸收峰.通过高效液相色谱分离得到1种刚果红降解产物,用质谱和傅立叶红外光谱鉴定,发现该产物相对分子量为184.2,主要官能团为-C6H4-和芳基-NH2,结合刚果红结构和该产物的核磁共振波谱推断其为联苯胺;并且随着降解时间的延长,联苯胺逐渐被降解.     

Effects of glucose on the decolorization of Reactive Black 5 by yeast isolates

The cometabolic roles of glucose were investigated in decolorization of an azo dye, Reactive Black 5, by yeast isolates, Debaryomyces polymorphus and Candida tropicalis. The results indicated that the dye degradation by the two yeasts was highly associated with the yeast growth process and glucose presence in the medium. Color removal of 200 mg dye/L was increased from 76.4% to 92.7% within 60 h to 100% within 18-24 h with the increase of glucose from 5 to 10 g/L, although the activity of manganese dependent peroxidase （MnP） decreased by 2-8 times in this case. Hydrogen peroxide of 233.3 μg/L was detected in 6 h in D. polymorphus culture. The cometabolic functions of glucose and hydrogen peroxide could be also confirmed by the further color removals of 95.8% or 78,9% in the second cycle of decolorization tests in which 7 g glucose/L or 250 μg H202/L was superadded respectively together with 200 mg dye/L.     

正交实验选择嗜碱细菌降解木质素的金属离子最优综合培养条件

本文在复合碳源的碱性液体培养条件下 (p H≈ 10 .5 ) ,用正交实验法对嗜碱木质素降解细菌 6号菌株降解木质素的金属离子培养条件进行了优化。结果表明 ,各金属离子对 6号菌株产生的木质素降解酶 L ac-case和 Mn P以及木质素降解率都有一定促进作用。分析正交实验结果表明 ,其中 Mn2 +和 Cu2 +是最主要的影响因素 ,各种金属离子的最佳综合水平为 :Mn SO4 · H2 O0 .7g/ L、Cu SO4 · 5 H2 O1m g/ L、Fe SO4 · 7H2 O5 mg/L、Zn SO4 · 7H2 O 30 m g/ L、Ca Cl2 · 2 H2 O 5 0 m g/     

Effects of culture conditions on ligninolytic enzymes and protease production by Phanerochaete chrysosporium in air

The production of ligninolytic enzymes and protease by Phanerochaete chrysosporium was investigated under different culture conditions. Different amounts of medium were employed in free and immobilized culture, together with two kinds of medium with different C/N ratios. Little lignin peroxidase （LIP） （〈 2 U/L） was detected in free culture with nitrogen-limited medium （C/N ratio： 56/2.2, in mmol/L）, while manganese peroxidase （MnP） maximum activity was 231 and 240 U/L in 50 and 100 ml medium culture, respectively. Immobilized culture with 50 ml nitrogen-limited medium gave the highest MnP and LiP production with the maximum values of 410 and 721 U/L separately on the day 5; however, flasks containing 100 ml nitrogen-limited medium only produced less MnP with a peak value of 290 U/L. Comparatively, carbon-limited medium （C/N ratio： 28/44, in mmol/L） was adopted in culture but produced little MnP and LiE Medium type had the greatest impact on protease production. Large amount of protease was produced due to glucose limitation. Culture type and medium volume influence protease activity corporately by affecting oxygen supply. The results implied shallow immobilized culture was a possible way to gain high production of ligninolytic enzymes.     

黄曲霉A5p1脱色类黑精及其机理初探:葡萄糖氧化酶的作用

类黑精是一种高分子难降解色素污染物质,在糖蜜酒精废水中大量存在.选取在前期研究中对糖蜜酒精废水具有较好脱色作用的黄曲霉(Aspergillus flavus)A5p1(保藏号CGMCC.4292),以合成的类黑精为对象研究脱色机理,试图为实际废水的生物脱色提供理论基础.结果显示,此菌株对合成类黑精具有生物吸附和生物降解的双重作用,以后者为主,最高脱色率可达65%.相较于漆酶、依赖/不依赖于锰的过氧化物酶等常规氧化脱色酶,此菌株中的产过氧化氢酶对脱色起主要作用,而且脱色过程中作为产过氧化氢酶之一的葡萄糖氧化酶与类黑精脱色率之间存在着一定的正相关性.添加葡萄糖氧化酶激活剂和抑制剂进行验证,发现葡萄糖氧化酶酶活力、过氧化氢产量和脱色率三者之间呈相关性变化.初步推测,葡萄糖氧化酶是黄曲霉A5p1生物降解脱色类黑精的关键酶之一.     

毒死蜱降解细菌XZ-3的分离及降解特性研究

从某农药厂排污口的污泥中采样,经驯化富集后筛选到1株能高效降解毒死蜱的细菌XZ-3,经过对其形态、生理生化特征及16S rDNA的分析,鉴定该菌株为节杆菌属（Arthrobactersp.）.测定了不同碳源、pH、温度及毒死蜱浓度对细菌降解能力和生长量的影响,以培养液在波长400 nm下的光密度值表示细菌生长量,液体介质中毒死蜱的提取采用漩涡振荡提取法,提取溶剂为石油醚,毒死蜱的测定采用紫外分光光度法.结果表明,该菌株24 h内对100 mg·L^-1毒死蜱的降解率高达86.8%;在外加碳源浓度为0.3%时降解率最大;细菌的生长量随着外加碳源浓度的升高而增加;pH在偏酸和偏碱性的条件下降解率较大,pH 9.0时达到最大,细菌的生长量在pH 8.0～10.0偏碱性的条件下较大;在毒死蜱浓度为100　mg·L^-1时降解率最大,该菌具有较强的抗药性,当毒死蜱浓度达到1?000　mg·L^-1时仍能生长,细菌的生长量在800　mg·L^-1时达到最大,绝对去除量随毒死蜱浓度的提高而增加;细菌的生长和降解需要适宜的温度,30℃培养时,降解率和生长量最大.本研究提出了细菌XZ-3生长和降解毒死蜱的最佳培养条件,可为农药污染治理及生产污水处理提供理论依据.     

Efficient purification of Al30 by organic complexation method

A method was developed for preparing high purity Al₃₀O₈(OH)₅₆(H₂O)₂₄^18 + (Al₃₀) through elimination of impurities by complexation. Polyaluminum chloride II (PAC₃₀) with Al_c content of 75% was adopted as the source of Al₃₀. The PAC₃₀ was prepared under conditions of total aluminum concentration 0.1?mol/L and OH^?/Al ratio 2.2 to obtain the highest content of Al₃₀. A precipitation/metathesis method, organic solvent precipitation method and organic complexation method were examined to separate and purify Al₃₀. It was found that only by the organic complexation method could high purity Al₃₀ products be obtained in large yield economically. In the experiments, benzoic acid was used as the coordinating reagent to decompose the main impurity AlO₄Al₁₂(OH)₂₄(H₂O)₁₂^7 + (Al₁₃), and the Al₃₀ product could be obtained by precipitation and metathesis operations. It was noteworthy that the decomposition of impurities by benzoic acid could be completed in 2?hr. The Al₃₀ product was characterized by Ferron assay, ²⁷Al-NMR, SEM, XRD and TGA. The results showed that the purity of the Al₃₀ product could exceed 92%.     

Optimizing aerobic biodegradation of dichloromethane using response surface methodology

Response surface methodology (RSM) was employed to evaluate the optimum aerobic biodegradation of dichloromethane (DCM) in pure culture. The parameters investigated include the initial DCM concentration, glucose as an inducer and hydrogen peroxide as terminal electron acceptor (TEA). Maximum aerobic biodegradation efficiency was predicted to occur when the initial DCM concentration was 380 mg/L, glucose 13.72 mg/L, and H2O2 115 mg/L. Under these conditions the aerobic biodegradation rate reached up to 93.18%, which was significantly higher than that obtained under original conditions. Without addition of glucose, degradation efficiencies were 6 80% at DCM concentrations < 326 mg/L. When concentrations of DCM were more than 480 mg/L, the addition of hydrogen peroxide did not help to significantly increase DCM degradation efficiency. When DCM concentrations increased from 240 to 480 mg/L, the overall DCM degradation efficiency decreased from 91% to 60% in the presence of H2O2 for 120 mg/L.     

Pilot regulation of MnP-SA for treating PTA wastewater

In the pilot study of treating the purified terephthalic acid(PTA) wastewater with the functional Strain Fhhh in the carrier activated sludge process(CASP), the ratio of COD:TN:TP and the concentrations of Cu, Mn, Se and Zn were controlled to improve the manganese peroxidase(MnP) levels for increasing the treatment efficiency. When the ratio of COD:TN:TP was 100:0.36:0.15 and the concentrations of Cu, Mn, Se and Zn were 0.54, 5.07, 0.00 and 0.08 mg/L, the MnP specific activity(MnP-SA) reached 689 U/L, and the sludge loading rate to COD(SLRC) was 1.09 d^-1, which was 4--7 fold of that in other processes reported. The data indicated that improving MnP level could enhance the degradability of Fhhh. And the potentials of Fhhh and CASP will be also discussed in this paper.     

黄曲霉A5p1对偶氮染料的降解特性和降解途径

以黄曲霉菌株A5p1为生物材料,研究其脱色染料的广谱性,并选择偶氮染料直接蓝71（DB71）为模型底物,探讨脱色特性及降解产物.该菌株对15种染料的脱色测试结果表明,染料浓度为100mg/L时脱色效率为61.7%~100%.该菌对偶氮染料DB71具有生物吸附和生物降解的双重作用,在pH值7.0,温度30℃,染料浓度300mg/L,蔗糖为碳源时对DB71 脱色率为100%.酶分析显示葡萄糖氧化酶和锰过氧化物酶参与染料的降解.FTIR、GC-MS和LC-MS分析确定代谢终产物为萘胺、叠氮萘、2-羟基-6-草酰-苯甲酸和1-萘酚.     

外加碳源对木糖氧化无色杆菌降解(艹屈)的影响

采用本实验室自行筛选的(艹屈)高效降解菌(木糖氧化无色杆菌)降解多环芳烃(艹屈),考察了葡萄糖、维生素C、邻苯二甲酸、丙酮酸、α-酮戊二酸、表面活性剂吐温-80等外加碳源对降解效果的影响。结果表明,适量添加葡萄糖、维生素C、丙酮酸、α-酮戊二酸和吐温80对的生物降解均有显著的刺激作用,邻苯二甲酸对木糖氧化无色杆菌降解(艹屈)不具有促进作用。反应体系中加入5mg/L维生素C时,木糖氧化无色杆菌对(艹屈)降解效率为91%。分别加入10mg/L的丙酮酸和α-酮戊二酸时,木糖氧化无色杆菌对(艹屈)降解效率均在90%以上。当加入200mg/L的吐温80时,木糖氧化无色杆菌对(艹屈)降解效率达到96%。