Characterization of the first step involved in enzymatic pathway for microcystin-RR biodegraded by Sphingopyxis sp. USTB-05

Enzymes encoded by genes biodegrading microcystins (MCs) can help reveal the function of genes and biodegradation pathway of MCs. Here the first and important gene (USTB-05-A, 1,008 bp) involved in biodegradation of microcystin-RR (MC-RR) was cloned from Sphingopyxis sp. USTB-05 and firstly expressed in Escherichia coli BL21 (DE3) with an expression vector of pGEX4T-1 successfully. The nucleotide sequences of cloned USTB-05-A possessed 92.5% homology to that of mlA reported in Sphingomonas sp. strain ACM-3962. The deduced amino acid sequences containing the cleavage sites of 26th (alanine) and 27th (leucine) showed 83% identical to that of MlrA. The cell-free extract (CE) of recombinant E. coli BL21 (DE3) containing USTB-05-A had high activity for biodegrading MC-RR. Initial MC-RR of 40 mg L⁻¹ was completely biodegraded under total protein of 350 mg L⁻¹ within 0.25 h. A product derived from MC-RR appeared distinctly with the decrease of MC-RR peak on the profile of HPLC. The product (m/z 1056.5) had molecular weight of 18 higher than that of MC-RR (m/z 1038.7). The findings provided the positive evidences that biodegradation of MC-RR began with the breakage of cyclic MC-RR and then it was converted to linear MC-RR as the first product catalyzed by first enzyme of Sphingopyxis sp. USTB-05.     

Citrobacter sp.CK3对活性红KN-3B的脱色反应动力学研究

采用脱色菌Citrobacter sp. CK3,以活性红KN-3B染料为处理对象,在厌氧批式反应条件下,系统考察了pH值,温度和染料浓度对脱色反应速度的影响;通过动力学模拟及反应过程中染料的UV-Vis扫描图分析,探讨了脱色反应机理。结果表明:Citrobacter sp. CK3对活性红KN-3B的脱色反应的适宜pH为7～9;脱色反应速度在温度为32℃时达到最大。染料初浓度从57 mg/L逐渐增大到458 mg/L时脱色率逐渐降低。脱色过程中染料的偶氮键发生断裂,脱色反应符合二级反应动力学。     

畜禽养殖污水中高效氨氮降解菌的筛选、鉴定及生长条件研究

从养殖污水中分离纯化得到1株高效氨氮降解菌,对其进行形态特征、生理生化、16S rDNA序列分析以及最佳生长条件研究,并将菌株投入养殖污水降解污水中的氨氮。结果表明：AN4菌株在NH4＋-N初始质量浓度为50 mg/L的条件下,24 h内的氨氮降解率为92.5%;初步鉴定该菌株为苍白杆菌属（Ochrobactrum sp.）,菌株的16S rDNA序列在Gen-Bank的登录号为GU345782;AN4菌株在装液量为60.64 mL,pH为7.06,葡萄糖为6.0 g/L的条件下培养,菌株的降解率可以达到94.28%;菌株对养殖污水中氨氮的降解率为12.3%。苍白杆菌菌属能够降畜禽养殖污水中的氨氮还未见报道,AN4菌株的筛选获得为生物降解养殖污水中的氨氮又提供了一种新型菌株。     

Toxicity of various anions associated with methoxyethyl methyl imidazolium-based ionic liquids on Clostridium sp

We investigated the effects on the growth of the anaerobic bacterium, Clostridium sp., of the ionic liquid, 1-methoxyethyl-3-methyl imidazolium [MOEMIM]⁺, derived from imidazolium cation and paired with one of a variety of counter-ions, viz., tetrafluoroborate [BF₄]^-, hexafluorophosphate [PF₆]^-_, trifluoroacetate [CF₃COO]^-, bis(trifluoromethane)sulfonamide [Tf₂N]^-, methane sulfonate [OMS], and 1-butyl-3-methyl imidazolium tetrafluoroborate [BMIM][BF₄]. These anions, in association with [MOEMIM]⁺ lowered the growth rate of the bacterium, showing the following trend: [Tf₂N]^- ≧ [PF₆]^- > [BF₄]^- > [CF₃COO]^- > [OMS]⁻. Anions incorporating fluorine were more toxic than those without it, and their toxicity rose with an increase in the number of fluorine atoms. Also, [MOEMIM]⁺[BF₄]^- was less toxic than [BMIM]⁺[BF₄]^-, probably due to the presence of a methoxyethyl functional group integrated in the cation side chain.     

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%。     

Relationship between chromium(VI) resistance and extracellular polymeric substances (EPS) concentration by some cyanobacterial isolates

BACKGROUND, AIM, AND SCOPE: Chromium(VI) resistance and its association with extracellular polymeric substance (EPS) concentration in cyanobacteria was investigated. Increased EPS concentration was associated with Cr(VI) resistance. The most resistant isolate, Chroococcus sp. H(4), secreted the most EPS (427 mg/L). MATERIALS AND METHODS: EPS concentration of the two most resistant isolates (Chroococcus sp. H(4) and Synechocystis sp. S(63)) was investigated following exposure to 15 and 35 ppm Cr(VI). The composition of EPS produced by Chroococcus sp. H(4) following exposure to 10 ppm Cr(VI) was analyzed using high-performance liquid chromatography. Control EPS was composed of glucose (99%) and galactronic acid (1%); in the presence of 10 ppm Cr(VI), EPS composition changed to glucose (9%), xylose (75%), rhamnose (14%), and galacturonic acid (2%). RESULTS AND DISCUSSION: Results indicated that (1) exposure to elevated concentrations of Cr(VI) affected the composition of EPS produced by Chroococcus sp. H(4), and (2) there was a correlation between Cr(VI) resistance and EPS concentration in some cyanobacteria.     

Degradation of poly(3-hydroxybutyrate), PHB,by bacteria and purification of a novel PHB depolymerase fromComamonas sp.

Bacteria capable of growing on poly(3-hydroxybutyrate), PHB, as the sole source of carbon and energy were isolated from various soils, lake water, activated sludge, and air. Although all bacteria utilized a wide variety of monomeric substrates for growth, most of the strains were restricted to degrade PHB and copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate, P(3HB-co-3HV). Five strains were also able to decompose a homopolymer of 3-hydroxyvalerate, PHV. Poly(3-hydroxyoctanoate), PHO, was not degraded by any of the isolates. One strain, which was identified asComamonas sp., was selected, and the extracellular depolymerase of this strain was purified from the medium by ammonium sulfate precipitation and by chromatography on DEAE-Sephacel and Butyl-Sepharose 4B. The purified PHB depolymerase was not a glycoprotein. The relative molecular masses of the native enzyme and of the subunits were 45,000 or 44,000, respectively. The purified enzyme hydrolyzed PHB, P(3HB-co-3HV), and—at a very low rate—also PHV. Polyhydroxyalkanoates, PHA, with six or more carbon atoms per monomer or characteristic substrates for lipases were not hydrolyzed. In contrast to the PHB depolymerases ofPseudomonas lemoignei andAlcaligenes faecalis T₁, which are sensitive toward phenylmethylsulfonyl fluoride (PMSF) and which hydrolyze PHB mainly to the dimeric and trimeric esters of 3-hydroxybutyrate, the depolymerase ofComamonas sp. was insensitive toward PMSF and hydrolyzed PHB to monomeric 3-hydroxybutyrate indicating a different mechanism of PHB hydrolysis. Furthermore, the pH optimum of the reaction catalyzed by the depolymerase ofComamonas sp. was in the alkaline range at 9.4.     

酵母菌与两种硫杆菌复合对污泥中三价铬的去除

研究了氧化亚铁硫杆菌LX5(Thiobacillus ferrooxidans LX5)、氧化硫硫杆菌TS6(Thiobacillus thiooxidans TS6)和耐酸性酵母菌R30(Rhodotorula sp.R30)对重金属铬(Cr³⁺)的耐受性.结果表明,700mg/L的Cr³⁺对硫杆菌LX5、TS6的生长和氧化活性影响不大,但Cr³⁺浓度大于500mg/L时明显抑制酵母菌R30的生长.酵母菌R30与硫杆菌LX5和TS6复合能明显加速污泥淋滤的进程,最佳复合比为酵母菌R30接种量2.0%,硫杆菌LX5和TS6接种量10%.分别用含酵母菌R30数量为10⁴个/mL和102个/mL的酸化污泥作接种物进行污泥淋滤,发现淋滤过程中pH值下降的速度没有明显差异,而在淋滤起始时添加2.0%的酵母菌R30,则淋滤反应提前36h结束.由此可见,在其它条件相同的污泥淋滤中,污泥中所含耐酸性酵母菌的数量是加快淋滤的关键.     

一种水培植物过滤法的除藻性能

运用隔离生态水区(mesocosm)的手法模拟再现小型封闭水体环境,对水培植物过滤法(HBFM)去除藻类的特性进行了研究.结果表明,藻类去除率平均可达61.1%,对有毒藻类Microcystis sp.的去除效果尤其明显.通过建立生态数学模型并进行计算机模拟,得出在循环比(日处理水量/封闭水体容量)为 0.2 的条件下,应用 HBFM 在 45d 内就可以使封闭水体的 Chl.a 浓度从 100μg/L 降低到 10μg/L .     

红球菌CDT3降解氯氰菊酯的特性及途径

以一株降解氯氰菊酯的红球菌(Rhodococcussp.)CDT3为材料,研究了CDT3降解氯氰菊酯的最适条件(温度、pH值、接种量、培养时间、添加营养物).结果表明,在温度30℃,pH8.0时,CDT3对氯氰菊酯的降解率最高,添加少量葡萄糖、蛋白胨、酵母汁均能促进它对氯氰菊酯的降解.研究了CDT3降解氯氰菊酯的代谢途径,通过GC-MS分析,证实氯氰菊酯被CDT3降解后的产物是3-苯氧基苯甲酸(3-PBA)和二氯菊酸(DCVA),推测氯氰菊酯是由CDT3产生的羧酸酯酶降解的,作为验证,提取了CDT3的粗酶液,并且通过聚丙烯酰胺凝胶电泳(PAGE)检测了羧酸酯酶.