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).     

呋喃丹降解菌CFDS-1的筛选鉴定及降解特性

为有效治理呋喃丹的污染,从受呋喃丹长期污染的土壤中分离筛选到一株高效降解呋喃丹的菌株CFDS-1,经形态、生理生化、16S rDNA(GenBank accession No.AY702969)同源性及系统发育地位等分析,将其初步鉴定为Sphingomonas sp.当接种量为5%时,CFDS-1能在48 h内降解100 mg L-1的呋喃丹,对于高达300 mg L-1的呋喃丹依然有降解效果;CFDS-1对呋喃丹的降解率与起始接种量呈正相关;降解呋喃丹的最适pH是8.0～9.0;在20～42℃范围内,温度对CFDS-1降解呋喃丹没有显著影响;该菌在250 mL三角瓶中装液量为100 mL时,对呋喃丹的降解效果最好.土壤实验表明,该菌株同样能有效地降解土壤中的呋喃丹残留.     

邻苯二酚2,3-双加氧酶基因克隆、定位和高效表达

采用特异性引物，以菲、芘降解菌株ZL5的代谢性质粒为模板，扩增出邻苯二酚2，3—双加氧酶(C23O)基因，将该基因和表达载体pET—30a( )连接，转化E.coli JM109(DE3)，获得了高效表达的转化子，SDS—PAGE结果表明，转化子的C23O蛋白不仅在细胞内存在，而且能被分泌到胞外，薄层扫描显示，转化子细胞内和细胞外表达蛋白总量占细胞总蛋白的42％，酶活分析表明，分布在转化子细胞内、外的表达蛋白都具有较高的C23O比活力，Southern杂交将菌株ZL5的C23O基因定位在内生质粒的不同酶切片段上。图5表1参12。     

固定化Acinetobacter sp. XA05和Sphingomonas sp. FG03降解苯酚

从活性污泥和受苯酚污染的土壤中分离出的菌株XA05和FG03均具有很强的苯酚生物降解能力。16srDNA序列分析表明,XA05和FG03菌株分别属于不动杆菌属(Acinetobacter sp.)和鞘氨醇单胞菌属(Sphin-gomonas sp.)。实验结果表明,在苯酚初始质量浓度为800.0mg/L、培养时间为35h的条件下,自由悬浮细胞和固定化细胞的苯酚降解率均高于95.0%。     

溴氨酸降解菌株的分离鉴定及特性研究

从污泥样品中分离得到一株溴氨酸降解菌,该菌株能够以溴氨酸为唯一碳、氮源及能源生长.通过形态、生理生化特性分析以及对16SrDNA序列进行同源比较,鉴定该菌株属于鞘氨醇单胞菌属,定名为Sphingomonasxenophaga .菌株的最适降解与生长条件为:pH =7 0 ,温度3 0℃,转速15 0r·min- 1 ,接种量8% .在最佳条件下,溴氨酸的降解率可达90 %以上,菌株可耐受的溴氨酸浓度为10 0 0mg·L- 1 .通过分析菌株对水杨酸、邻苯二酚及邻苯二甲酸的利用情况,推测该菌株可能通过邻苯二酚的代谢途径降解溴氨酸.     

一株菲高效降解菌的环境适应性研究

生物降解是多环芳烃从环境中去除的主要途径,菲是一种典型的三环芳烃。本研究考察了一株能高效降解多环芳烃菲的鞘氨醇单胞菌GY2B在含河沙环境及不同盐度的人工海水环境中的生长特性与降解菲的情况。结果表明：河沙的加入对菌株GY2B的生长及其高效降解菲的性能均无明显影响,65 h可将起始浓度为100 mg/L的菲降解99.5%以上;而经过驯化后在添加85%人工海水的条件下该菌也仍可正常生长并高效降解菲,66 h可将起始浓度为100 mg/L的菲几乎完全降解。本研究结果可为菌株GY2B在受多环芳烃污染的河滩、河口及近海海洋环境修复中的应用提供参考依据。     

固定化菌降解溴氨酸的特性及其动力学研究

鞘氨醇单胞菌QYY菌能以溴氨酸为唯一碳源、氮源和能源生长。考察了用海藻酸钙包埋该菌细胞对溴氨酸废水进行处理的降解和动力学过程。确定了固定化鞘氨醇单胞菌QY(Y以下简称固定化菌)处理溴氨酸废水的最适条件为:接种量10mL固定化菌,摇床转速100r/min,pH7,温度30℃。最佳降解条件下,固定化菌降解溴氨酸的过程符合Haldane模型。     

鞘氨醇单胞菌完整细胞对溴氨酸的好氧降解

文章考察了鞘氨醇单胞菌QYY完整细胞对溴氨酸的好氧降解条件,并对其降解途径进行了分析。研究表明,该完整细胞在最佳条件为:温度30℃、初始pH7.0～8.0、摇床转速150r/min,磷酸缓冲溶液浓度为0.01～0.04mol/L时将溴氨酸完全脱色后产生3种产物:2-氨基-3-羟基-5-溴苯磺酸、2-氨基-4-羟基-5-溴苯磺酸为终产物;邻苯二甲酸为中间产物。其脱色过程中,大部分溴氨酸直接降解为终产物,仅有一小部分溴氨酸在降解过程中形成邻苯二甲酸。邻苯二甲酸则通过苯甲酸代谢途径开环。     

多环芳烃降解菌ZL5分离鉴定及其降解质粒

通过选择性富集培养，从辽河油田石油污染土壤中分离到一株多环芳烃(PAHs)降解菌ZL5．它能以菲和芘为唯一碳源生长，但是不能利用萘．16S rDNA核苷酸序列分析结果表明，ZL5属于变形细菌α亚类中的鞘氨醇单胞菌属．该菌株含有一个大小约为60kb的质粒．丝裂霉素C消除实验表明，随着质粒的丢失，菌株利用菲和芘的能力也丧失．用电转化和氯化铷转化法分别将菌株ZL5的质粒导人大肠杆菌JM109和DH5α中，随着质粒的获得，这些转化子获得了降解菲和芘的能力．本研究结果表明，鞘氨醇单胞菌ZL5降解PAHs的功能和质粒有关。