Enhanced bio-decolorization of 1-amino-4-bromoanthraquinone-2-sulfonic acid by Sphingomonas xenophaga with nutrient amendment

Bacterial decolorization of anthraquinone dye intermediates is a slow process under aerobic conditions. To speed up the process, in the present study, effects of various nutrients on 1-amino-4-bromoanthraquinone-2-sulfonic acid (ABAS) decolorization by Sphingomonas xenophaga QYY were investigated. The results showed that peptone, yeast extract and casamino acid amendments promoted ABAS bio-decolorization. In particular, the addition of peptone and casamino acids could improve the decolorization activity of strain QYY. Further experiments showed that L-proline had a more significant accelerating effect on ABAS decolorization compared with other amino acids. L-Proline not only supported cell growth, but also significantly increased the decolorization activity of strain QYY. Membrane proteins of strain QYY exhibited ABAS decolorization activities in the presence of L-proline or reduced nicotinamide adenine dinucleotide, while this behavior was not observed in the presence of other amino acids. Moreover, the positive correlation between L-proline concentration and the decolorization activity of membrane proteins was observed, indicating that L-proline plays an important role in ABAS decolorization. The above findings provide us not only a novel insight into bacterial ABAS decolorization, but also an L-proline-supplemented bioaugmentation strategy for enhancing ABAS bio-decolorization.     

Quinone-mediated decolorization of sulfonated azo dyes by cells and cell extracts from Sphingomonas xenophaga

The effects of various quinone compounds on the decolorization rates of sulfonated azo dyes by Sphingomonas xenophaga QYY were investigated. The results showed that anthraquinone-2-sulfonate (AQS) was the most effective redox mediator and AQS reduction was the rate-limited step of AQS-mediated decolorization of sulfonated azo dyes. Based on AQS biological toxicity tests, it was assumed that AQS might enter the cells to kill them. In the cytoplasmic extracts from strain QYY, AQS effectively increased decolorization rates of sulfonated azo dyes than other quinone compounds. In addition, we found a NADH/FMN-dependent AQS reductase using nondenaturing polyacrylamide gel electrophoresis (Native-PAGE).     

菌株HP3对溴胺酸的降解机理

通过正交试验,得到菌株HP3降解溴胺酸最佳条件为温度30℃,pH值7.0,摇床转速160r/min,培养基中不含NaCl有利于溴胺酸的降解.在此条件下,溴胺酸的降解遵从负指数模式.葡萄糖和硫酸铵的加入显著地提高菌体降解溴胺酸的速度.不同金属化合物对菌株HP3降解溴胺酸有不同程度的抑制作用,其中以HgCl₂和AgNO₃最显著.菌株HP3能降解化合物蒽醌、1,4,5,8-四羟基蒽醌和1-氨基蒽醌-2-磺酸钠,表明该菌株对底物的降解没有严格的专一性.溴胺酸经菌株HP3作用后吸收光谱发生明显变化.TOC分析表明,菌株HP3将溴胺酸的蒽醌环破坏后利用其中的一部分有机碳.液相色谱检测有中间产物邻苯二甲酸生成.溴胺酸降解终产物分子量为289和290.     

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.     

醌介导染料脱色菌株的分离鉴定及特性

实验分离获得1株能够利用醌化合物使磺酸化偶氮染料脱色的菌株JL,通过形态特征、16S rDNA与16S-23S区间序列分析表明,该菌株为蜡状芽孢杆菌(命名为Bacillus cereus JL).菌株JL使酸性大红3R脱色的最佳条件为葡萄糖浓度1g/L, pH值为5~7,温度30℃,接种量0.25g/L.蒽醌-2-磺酸(AQS)、蒽醌-2,6-二磺酸(AQDS)和2-羟基-1,4-萘醌(Lawsone)均能显著提高酸性大红3R的脱色速率,其中AQS的促进作用最为明显.研究发现, 0.1mmol/L AQS能够使菌株JL对2.0mmol/L酸性大红3R保持较高的脱色速率,而且能使多种偶氮染料脱色,表现出较好的底物广谱性.利用高效液相色谱-质谱鉴定了AQS介导的酸性大红3R脱色产物,表明酸性大红3R的偶氮键发生断裂, AQS在这一过程中仅起到电子传递的作用.     

Trametes sp.SQ01和Chaetomium sp.R01混合培养对四种染料的脱色

利用本实验室新构建的白腐菌Trametes sp.SQ01和毛壳菌Chaetomium sp.R01混合培养体系,对刚果红、酸性红、橙黄G和溴酚蓝4种染料进行了脱色研究.结果表明,SQ01与R01混合培养所产生的锰过氧化物酶(MnP)酶活比菌株SQ01单独培养时提高了约5.5倍.菌株R01的接种量、接种时间对混合培养中...     

堆肥腐殖酸形成及其电化学活性

为探究堆肥腐殖酸形成过程及腐殖酸的氧化还原性能,以城市生活垃圾不同堆肥阶段提取的腐殖酸样品为研究对象,运用现代色谱学技术、光谱学方法和电化学分析,研究了堆肥过程不同形态氮和碳官能团形成腐殖酸的特征,探究了堆肥腐殖酸的氧化还原性能及影响因素. 结果表明:堆肥腐殖酸中50%~75%的氮为氨基酸态氮,检出的15种氨基酸中酸性氨基酸(Asp和Glu)含量(高于100 mg/g)最高、丝氨酸(Ser)含量(3.78 mg/g)最低. 与堆肥腐殖酸中其他组分和官能团相比,氨基酸易被生物降解和利用,其含量随着发酵的进行呈下降趋势,后期氨基酸态氮占比逐渐降低. 腐殖酸中碳有多种形态,包括苯环、羧基、脂肪族等多种官能团,随着堆肥发酵的进行,木质纤维素降解形成腐殖酸,脂肪族官能团亦被氧化降解;与之相反的是,腐殖酸中羧酸类、醛、酮及苯环等官能团不断增加,导致腐殖酸氧化还原性能增强. 氧化和还原过程中,腐殖酸得失电子均会造成部分官能团结构破坏,大分子腐殖酸被降解和转化为小分子有机物. 研究显示,堆肥腐殖酸活性较高,可利用腐殖酸修复环境过程,但需要适当补充活性腐殖酸,防止腐殖酸因修复氧化还原过程导致降解后只能作为碳源.      

固定化蒽醌对偶氮染料生物降解促进作用研究

采用非水溶性蒽醌固定化技术对偶氮染料生物降解促进作用进行了研究,对比了海藻酸钙,聚乙烯醇-硼酸,聚乙烯醇-海藻酸钙和琼脂4种固定化技术,探讨了溶解氧对脱色过程的影响和固定化蒽醌系统脱色广谱性.研究结果表明,固定化蒽醌可提高多种偶氮染料生物厌氧脱色速度1.5～2倍和降低偶氮染料脱色过程氧化还原电位-10～-15 mV;经4次循环使用后,其加速作用仍保持在90%以上;固定化蒽醌微生物系统具有很强抗氧冲击能力.     

1株异养硝化菌胞外聚合物的研究

研究异养硝化菌(Acinetobacter sp.YY-5)在不同生长时期胞外聚合物(EPS)组成成分及其变化规律.利用热处理提取YY-5菌的EPS,分析该菌株在不同生长时期EPS组分,包括蛋白质、总糖、核酸和游离氨基酸等.并进一步把蛋白质水解成氨基酸,分析其氨基酸的变化规律及其对蛋白质理化性质的影响.结果表明,该菌株具有较高含量的EPS,且其在稳定期时EPS含量最高.在不同时期分泌的EPS组分均以蛋白质为主.EPS中各组分含量及其变化规律与该菌生长状态密切相关,在该菌株生长过程中,EPS中各组分的变化情况:蛋白质从14.599 mg.g-1增加到28.489 mg.g-1后减少为15.139 mg.g-1;总糖从6.757 mg.g-1增加到10.199 mg.g-1后减少为7.857 mg.g-1.核酸从1.56 mg.g-1一直增加到6.287 mg.g-1;游离氨基酸从3.713 mg.g-1增加到4.374 mg.g-1后减少为1.299 mg.g-1.蛋白质水解后,其中极性氨基酸呈先增加后减少的趋势,非极性氨基酸一直呈增加趋势;在生长过程中一直以带负电的氨基酸占优.     

Biodegradation of Crystal Violet by Agrobacterium radiobacter

Agrobacterium radiobacter MTCC 8161 completely decolorized the Crystal Violet with 8 hr (10 mg/L) at static anoxic conditions. The decreased decolorization capability by A. radiobacter was observed, when the Crystal Violet concentration was increased from 10 to 100 mg/L. Semi-synthetic medium containing 1% yeast extract and 0.1% NH4Cl has shown 100% decolorization of Crystal Violet within 5 hr. A complete degradation of Crystal Violet by A. radiobacter was observed up to 7 cycles of repeated addition (10 mg/L). When the effect of increasing inoculum concentration on decolorization of Crystal Violet (100 mg/L) was studied, maximum decolorization was observed with 15% inoculum concentration. A significant increase in the activities of laccase (184%) and aminopyrine N-demethylase (300%) in cells obtained after decolorization indicated the involvement of these enzymes in decolorization process. The intermediates formed during the degradation of Crystal Violet were analyzed by gas chromatography and mass spectroscopy (GC/MS). It was detected the presence of N,N,N′,N′′-tetramethylpararosaniline, [N; N-dimethylaminophenyl] [N-methylaminophenyl] benzophenone, N; N-dimethylaminobenzaldehyde, 4-methyl amino phenol and phenol. We proposed the hypothetical metabolic pathway of Crystal Violet biodegradation by A. radiobacter. Phytotoxicity and microbial toxicity study showed that Crystal Violet biodegradation metabolites were less toxic to bacteria (A. radiobacter, P. aurugenosa and A. vinelandii) contributing to soil fertility and for four kinds of plants (Sorghum bicolor, Vigna radiata, Lens culinaris and Triticum aestivum) which are most sensitive, fast growing and commonly used in Indian agriculture.     

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.     

Decolorization of dye solutions with Ruditapes philippinarum conglutination mud and the isolated bacteria

Application of Ruditapes Philippinarum conglutination mud (RPM) for decolorizing synthetic dye solutions was studied. RPM showed good activity for decolorization of Methylene Blue, Crystal Violet, Malachite Green, and Ink Blue. The amount of the RPM had great effect on the decoloration rate of the dye solutions. However, the decoloration rate did not continue to increase when the amount of mud exceeded the optimum dose. The temperature of the dye solution had a remarkable effect on the decolorization rate of Ink Blue solution, but had little effect on the other three dye solutions. The initial pH of the dye solutions evidently affected the decolorization rate of Malachite Green solution, but had less effect on the other three. The decolorization rate of the dye solutions increased significantly with treatment time within 8 hr, but tended to be steady after 8 hr for Methylene Blue, Crystal Violet and Malachite Green solutions, and after 12 hr for Ink Blue solution. The decolorization efficiencies for the four dye solutions under the optimum conditions were all above 90%. Seventeen strains screened from RPM showed flocculation ability for kaolin clay suspension. Out of them, the flocculation rate of strain ZHT3-9 and strain ZHT4-13 were up to 88.14% and 86.01%, respectively. ZHT3-9 was studied, and its decolorization rate for Methylene Blue, Crystal Violet, and Malachite Green reached 90.02%, 89.21%, and 80.29%, respectively. By morphological, physiological and biochemical characteristics analysis and 16S rRNA sequencing, the strain ZHT3-9 was identified as Arthrobacter sp.     

菌株HX5对多种染料的吸附作用

研究了HX5生长菌体对分属活性、酸性、碱性、直接和分散5大类的26种染料的吸附性能．结果表明，HX5生长菌体可在5h内完全吸附直接染料和分散染料，碱性染料在生长菌体上完全吸附脱色的时间为12h，其次是活性染料，最不易吸附的为酸性染料．染料在生长菌体上的吸附由染料分子的结构和性质所决定，直接染料分子呈线性平面结构、疏水性较强，易于吸附，完全吸附脱色的时间为5h；分散性染料水溶性较差，在静电力的诱导之下即可在较短时间内完全吸附脱色；碱性染料分子带正电，与带负电荷的菌丝球表面异性电荷相吸从而发生吸附脱色；活性染料分子中的氨基质子化后与菌丝球表面相互吸引，氨基质子化的程度越高，吸附效果也就越好；酸性染料分子中氨基质子化程度受到结构本身的限制，以及不存在带正电荷的基团，因而吸附效果最差．     

黄杆菌(Flavobacterium sp.)对溴氨酸脱色的研究

从受污土壤中筛选出对溴氨酸具有高效脱色能力的黄杆菌BX26．通过监测液体培养基中细胞浊度和溴氨酸吸光度的变化,考察了碳源浓度、溴氨酸浓度和以溴氨酸为唯一碳、氮、硫源时对菌体生长和溴氨酸脱色的影响．结果表明,BX26对溴氨酸的脱色在于酶的催化作用．该脱色酶属诱导性胞外酶．酶的诱导和产生需要液体培养基中有足量的氮源和硫源,但对碳源没有要求．溴氨酸对菌体的生长具有明显的促进作用,同时菌株也表现出很高的溴氨酸脱色活性,可使高达1000mg/L的溴氨酸降解脱色．菌体不能以溴氨酸为唯一碳、氮、硫源,但可以溴氨酸的脱色产物为唯一碳源．     

Catalyzed degradation of disperse dyes by calcium alginate-pectin entrapped bitter gourd （Momordica charantia） peroxidase

Calcium-alginate pectin entrapped bitter gourd peroxidase (BGP) has been employed for the treatment of disperse dyes: Disperse Brown 1 (DB 1) and Disperse Red 17 (DR 17). Peroxidase alone was unable to decolorize DR 17 and DB 1. However, the investigated dyes were decolorized maximally by BGP in the presence of 0.2 mmol/L redox mediator, violuric acid (VA). A slow decrease in percent decolorization was observed when VA concentration was higher than 0.2 mmol/L which could likely be due to the high reactivity of its aminoxyl radical (> N–O.) intermediate, that might undergo chemical reactions with aromatic amino acid side chains of the enzyme thereby inactivating it. Maximum decolorization of the dyes was observed at pH 3.0 and 40°C within 2 hr of incubation. Immobilized peroxidase decolorized 98% DR 17 and 71% DB 1 using 35 U of BGP in batch process in 90 min. Immobilized enzyme decolorized 85% DR 17 and 51% DB 1 whereas soluble enzyme decolorized DR 17 to 48% and DB 1 to 30% at 60°C. UV-visible spectral analysis was used to evaluate the degradation of these dyes and their toxicity was tested by Allium cepa test. The generally observed higher stability of the bioaffinity bound enzymes against various forms of inactivation may be related to the specific and strong binding of enzyme with bioaffinity support which prevents the unfolding/denaturation of enzyme. Thus entrapped peroxidase was found to be effective in the decolorization of the investigated dyes.     

Decolorization of molasses melanoidins and palm oil mill effluent phenolic compounds by fermentative lactic acid bacteria

Lactobacillus plantarum SF5.6 is one of the lactic acid bacteria (LAB) that has the highest ability of molasses melanoidin (MM) decolorization among the 2114 strains of LAB. The strains were isolated from spoilage, pickle fruit and vegetable, soil and sludge from the wastewater treatment system by using technical step of enrichment, primary screening and secondary screening. This LAB strain SF5.6 was identified by 16S rDNA analysis and carbohydrate fermentation (API 50 CH). The top five LAB strains having high MM decolorization ( 55%), namely TBSF5.8-1, TBSF2.1-1, TBSF2.1, FF4A and SF5.6 were selected to determine the optimal condition. It was found that the temperature at 30°C under facultative conditions in GPY-MM medium (0.5% glucose, 0.1% peptone, 0.1% yeast extract, 0.1% sodium acetate, 0.05% MgSO4 and 0.005% MnCl2 in MM solution at pH 6) giving a high microbial growth and MM decolorization for all five strains. It was noticed that the decolorization of MM by LAB strains might be cell growth associated. L. plantarum SF5.6 grew rapidly within one day while the other strains took 2–3 days. This L. plantarum SF5.6 could rapidly decolorize MM to 60.91% without any lag phase, and it also had the ability to remove 34.00% phenolic compounds and 15.88% color from treated palm oil mill effluent.     

Decolorization of molasses melanoidins and palm oil mill e uent phenolic compounds by fermentative lactic acid bacteria

Lactobacillus plantarum SF5.6 is one of the lactic acid bacteria (LAB) that has the highest ability of molasses melanoidin (MM) decolorization among the 2114 strains of LAB. The strains were isolated from spoilage, pickle fruit and vegetable, soil and sludge from the wastewater treatment system by using technical step of enrichment, primary screening and secondary screening. This LAB strain SF5.6 was identified by 16S rDNA analysis and carbohydrate fermentation (API 50 CH). The top five LAB strains having high MM decolorization (> 55%), namely TBSF5.8-1, TBSF2.1-1, TBSF2.1, FF4A and SF5.6 were selected to determine the optimal condition. It was found that the temperature at 30°C under facultative conditions in GPY-MM medium (0.5% glucose, 0.1% peptone, 0.1% yeast extract, 0.1% sodium acetate, 0.05% MgSO4 and 0.005% MnCl2 in MM solution at pH 6) giving a high microbial growth and MM decolorization for all five strains. It was noticed that the decolorization of MM by LAB strains might be cell growth associated. L. plantarum SF5.6 grew rapidly within one day while the other strains took 2–3 days. This L. plantarum SF5.6 could rapidly decolorize MM to 60.91% without any lag phase, and it also had the ability to remove 34.00% phenolic compounds and 15.88% color from treated palm oil mill e uent.     

多金属氧酸盐电催化降解染料废水的研究

采用IR和XRD等手段对磷钼酸(PMo12)和3,3',5,5'-四甲基联苯胺(TMB)形成的电荷转移配合物(TMB)3PMo12进行表征,结果表明配合物中的杂多阴离子仍保留着Keggin结构.将配合物负载于γ-Al2O3上制备(TMB)3PMo12/γ-Al2O3负载型催化剂填充于电化学反应器中,考察电催化氧化体系降解酸性大红3R染料废水的效果.研究表明,(TMB)3PMo12/γ-Al2O3催化剂对酸性大红3R废水显示了良好的电催化活性,催化剂填充量为50g,负载量为0.6%时,在pH值为4,槽电压15.0V,曝气量0.14m3/h,极板间距3.0cm反应条件下,60min后,色度去除率达到79.7%,去除效果较传统二维平板和三维电解槽更高.     

Enterobacter sp. CV-v对甲基橙的脱色特性与条件优化

从浙江温州分离筛选到1株甲基橙高效脱色菌株CV-v,经16S rRNA基因序列分析表明,该菌株属于Enterobacter sp.属.单因素实验结果表明:当pH值在5.0~9.0之间时,培养48h以后,该菌株对甲基橙的脱色率均在80%以上;脱色的最适温度范围为30~40℃之间;所测碳源中的葡萄糖和麦芽糖、氮源中的牛肉膏和酵母粉对脱色的促进效果最为显著,且Ca2+和Mg2+也对脱色有显著的促进效应;此外,当接种量达到7%(V:V)以后,48h的脱色率即可达近100%.响应面设计实验结果显示,该菌株对甲基橙脱色的最优操作条件为:pH 8.0,葡萄糖1.7g/L,酵母粉3.0g/L,氯化镁3.0mmol/L及培养温度35.9℃.验证实验结果表明在最优条件下,该菌株在10h内对甲基橙的脱色率可达88.0%.总体而言,该菌株在偶氮染料脱色中的应用潜能较大.     

Cloning, expression in Escherichia coli, and enzymatic properties of laccase from Aeromonas hydrophila WL-11

A strain WL-11 with high laccase activity was isolated from activated sludge collected from the effluent treatment plant of a textile and dyeing industry.It was identified as Aeromonas hydrophila by physiological test and 16S rDNA sequence analysis.A gene encoding of laccase from a newly isolated Aeromonas hydrophila WL-11 was cloned and characterized.Nucleotide sequence analysis showed an open reading frame of 1605 bp encoding a polypeptide comprised of 534 amino acids.The primary structure of the enzyme predicted the structural features characteristic of other laccases,including the conserved regions of four histidine-rich copper-binding sites.The predicted amino acid sequence showed a high homology(more than 60%) with bacterial laccases in the genome and protein databases and the highest degree of similarity(61% identity) was observed with the multicopper oxidase of Klebsiella sp.601.When expressed in Escherichia coli,the recombinant enzyme was overproduced in the cytoplasm as soluble and active form.The purified enzyme had an optimum pH of 2.6 and 8.0 for ABTS(2,2'-azino-bis(3-ethylbenzthiazolinesulfonic acid) and DMP(2,6-dimethoxyphenol),respectively.The kinetic study on ABTS revealed a higher affinity of this enzyme to this substrate than DMP.