Cloning and expression of the first gene for biodegrading microcystin LR by Sphingopyxis sp. USTB-05

Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins (MCs) produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-05-A (HM245411) that is responsible for the first step in the biodegradation of microcystin LR (MC-LR) in E. coli DH5αup, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts (CE) of recombinant E. coli DH5αup containing USTB-05-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum (LC-MS), the enzyme encoded by gene USTB-05-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid (Adda) and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05.     

Carbon dioxide fixation by Chlorella sp. USTB-01 with a fermentor-helical combined photobioreactor

A promising microalgal strain isolated from fresh water, which can grow both autotrophically on inorganic carbon under lighting and heterotrophically on organic carbon without lighting, was identified as Chlorella sp. USTB-01 with the phylogenetic analysis based on 18S ribosomal ribonucleic acid (rRNA) gene sequences. In the heterotrophic batch culture, more than 20.0 g·L⁻¹ of cell dry weight concentration (DWC) of Chlorella sp. USTB-01 was obtained at day 5, and which was used directly to seed the autotrophic culture. A novel fermentor-helical combined photobioreactor was established and used to cultivate Chlorella sp. USTB-01 for the fixation of carbon dioxide (CO₂). It showed that the autotrophic growth of Chlorella sp. USTB-01 in the combined photobioreactor was more effective than that in the fermentor alone and the maximum DWC of 2.5 g·L⁻¹ was obtained at day 6. The highest CO₂ fixation of 95% appeared on day 1 in the exponential growth phases of Chlorella sp. USTB-01 and 49.8% protein was found in the harvested microalgal cells.     

鞘氨醇单胞菌USTB-05对微囊藻毒素的生物降解

研究了云南滇池水华蓝藻细胞中微囊藻毒素(MCs)的分析和鞘氨醇单胞菌USTB-05在细胞水平和酶水平下对MC-YR、RR和LR的生物降解.结果表明,云南滇池水华蓝藻细胞中MC-YR、RR和LR的含量分别为0.16, 0.96, 0.47mg/g.在初始浓度分别为19.5mg/L MC-YR、79.5mg/L MC-RR和43.6mg/L MC-LR下,鞘氨醇单胞菌USTB-05在2d内可将上述3种MCs全部降解,鞘氨醇单胞菌USTB-05粗酶液可以以更快的速率对MC-YR、MC-RR和MC-LR进行高效酶催化降解,在10h内可以将初始浓度分别为14.8mg/L MC-YR、28.4mg/L MC-RR和19.5mg/L MC-LR全部降解.同时发现了MC-YR降解过程中的2个中间和1个最终代谢产物.     

Adsorption of fluoride on clay minerals and their mechanisms using X-ray photoelectron spectroscopy

This research investigates the adsorption mechanisms of fluoride (F) on four clay minerals (kaolinite, montmorillonite, chlorite, and illite) under different F⁻ concentrations and reaction times by probing their fluoride superficial layer binding energies and element compositions using X-ray photoelectron spectroscopy (XPS). At high F⁻ concentrations (C ₀ = 5–1000 mg·L⁻¹), the amount of F⁻ adsorbed (Q _F), amount of hydroxide released by clay minerals, solution F⁻ concentration, and the pH increase with increasing C ₀. The increases are remarkable at C ₀>50 mg·L⁻¹. The QF increases significantly by continuously modifying the pH level. At C ₀<5–100 mg·L⁻¹, clay minerals adsorb H⁺ to protonate aluminum-bound surface-active hydroxyl sites in the superficial layers and induce F⁻ binding. As the C ₀ increases, F⁻, along with other cations, is adsorbed to form a quasi-cryolite structure. At C ₀>100 mg·L⁻¹, new minerals precipitate and the product depends on the critical Al³⁺ concentration. At [Al³⁺]>10^−11.94 mol·L⁻¹, cryolite forms, while at [Al³⁺]<10^−11.94 mol·L⁻¹, AlF₃ is formed. At low C ₀ (0.3–1.5 mg·L⁻¹), proton transfer occurs, and the F⁻ adsorption capabilities of the clay minerals increase with time.     

降解微囊藻毒素菌种的筛选和活性研究

研究了滇池底泥和表层水体中的微生物菌群降解微囊藻毒素(Microcystins, MCs)的能力差异,发现底泥中的微生物菌群对MCs有更强的生物降解能力.采用从滇池水华蓝藻细胞中提取提纯的微囊藻毒素作为微生物生长的唯一碳源和氮源,先后经过液体和固体培养基培养后,通过挑取单克隆菌落,分别从底泥中筛选出了能够降解MC-RR和LR的5种不同微生物菌种.其中筛选的菌种D降解MCs的能力最强,在3 d内可将初始浓度分别为60.1 mg·L^-1和38.7 mg·L^-1的MC-RR和LR全部降解,日均降解MC-RR和LR的速率分别高达20.0 mg·L^-1和12.9 mg·L^-1.     

布洛芬降解菌降解谱研究

以从西安污水处理厂筛选所得的3株布洛芬降解菌株I2(克雷伯氏菌)、I4(假单胞菌)和I14(不动杆菌)为研究对象,探索其生长特性及对布洛芬同系物降解的广谱性.结果表明,3株菌对7种选定的布洛芬同系物均有降解作用,但降解能力表现出一定的差异.I2、I4菌株的降解能力优于I14菌株,对7种布洛芬同系物的最高耐受浓度均在300 mg·L-1以上.3株降解菌都对布洛芬同系物中的间苯二酚有良好的耐受能力,最高耐受浓度均在1000 mg·L-1以上,I2菌株甚至达到2500 mg·L-1;3株降解菌对于邻苯二酚、对苯二酚的耐受程度最差,其中,I14菌株在对苯二酚达到100 mg·L-1时就会死亡.I2、I4菌株可作为布洛芬及其同系物降解的优良备选菌株,用于进一步研究多重污染治理的方式.     

Influence of aeration intensity on the performance of A/O-type sequencing batch MBR system treating azo dye wastewater

Among the numerous parameters affecting the membrane bioreactor (MBR) performance, the aeration intensity is one of the most important factors. In the present investigation, an anoxic/aerobic-type (A/O-type) sequencing batch MBR system, added anoxic process as a pretreatment to improve the biodegradability of azo dye wastewater, was investigated under different aeration intensities and the impact of the aeration intensity on effluent quantity, sludge properties, extracellular polymeric substances (EPS) amount generated as well as the change of permeation flux were examined. Neither lower nor higher aeration intensities could improve A/O-type sequencing batch MBR performances. The results showed 0.15 m³·h⁻¹ aeration intensity was promising for treatment of azo dye wastewater under the conditions examined. Under this aeration intensity, chemical oxygen demand (COD), ammonium nitrogen and color removal as well as membrane flux amounted to 97.8%, 96.5%, 98.7% and 6.21 L·m⁻²·h⁻¹, respectively. The effluent quality, with 25.0 mg·L⁻¹COD, 0.84 mg·L⁻¹ ammonium nitrogen and 8 chroma, could directly meet the reuse standard in China. In the meantime, the sludge relative hydrophobicity, the bound EPS, soluble EPS and EPS amounts contained in the membrane fouling layer were 70.3%, 52.0 mg·g⁻¹VSS, 38.8 mg·g⁻¹VSS and 90.8 mg·g⁻¹VSS, respectively, which showed close relationships to both pollutant removals and membrane flux.     

Migration of manganese and iron during the adsorption-regeneration cycles for arsenic removal

Fe-Mn binary oxide incorporated into porous diatomite (FMBO-diatomite) was prepared in situ and regenerated in a fixed-bed column for arsenite [As(III)] and arsenate [As(V)] removal. Four consecutive adsorption cycles were operated under the following conditions: Initial arsenic concentration of 0.1 mg·L⁻¹, empty bed contact time of 5 min, and pH 7.0. About 3000, 3300, 3800, and 4500 bed volumes of eligible effluent (arsenic concentration ⩽ 0.01 mg·L⁻¹) were obtained in four As (III) adsorption cycles; while about 2000, 2300, 2500, and 3100 bed volumes of eligible effluent were obtained in four As(V) adsorption cycles. The dissection results of FMBO-diatomite fixed-bed exhibited that small amounts of manganese and iron were transferred from the top of the fixed-bed to the bottom of the fixed-bed during As(III) removal process. Compared to the extremely low concentration of iron (<0.01 mg·L⁻¹), the fluctuation concentration of Mn²⁺ in effluent of the As(III) removal column was in a range of 0.01–0.08 mg·L⁻¹. The release of manganese suggested that manganese oxides played an important role in As(III) oxidation. Determined with the US EPA toxicity characteristic leaching procedure (TCLP), the leaching risk of As(III) on exhausted FMBO-diatomite was lower than that of As(V).     

Biosorption of Cr(III) from aqueous solution by freeze-dried activated sludge: Equilibrium, kinetic and thermodynamic studies

Batch biosorption experiments were conducted to remove Cr(III) from aqueous solutions using activated sludge from a sewage treatment plant. An investigation was conducted on the effects of the initial pH, contact time, temperature, and initial Cr(III) concentration in the biosorption process. The results revealed that the activated sludge exhibited the highest Cr(III) uptake capacity (120 mg·g⁻¹) at 45°C, initial pH of 4, and initial Cr(III) concentration of 100 mg·L⁻¹. The biosorption results obtained at various temperatures showed that the biosorption pattern accurately followed the Langmuir model. The calculated thermodynamic parameters, ΔGo° ( − 0.8–4.58 kJ·mol⁻¹), ΔH° (15.6–44.4 kJ·mol⁻¹), and ΔS° (0.06–0.15 kJ·mol⁻¹·K⁻¹) clearly indicated that the biosorption process was feasible, spontaneous, endothermic, and physical. The pseudo first-order and second-order kinetic models were adopted to describe the experimental data, which revealed that the Cr(III) biosorption process conformed to the second-order rate expression and the biosorption rate constants decreased with increasing Cr (III) concentration. The analysis of the values of biosorption activation energy (E _a = −7 kJ·mol⁻¹) and the intraparticle diffusion model demonstrated that Cr(III) biosorption was film-diffusion-controlled.     

微囊藻毒素降解菌的分子鉴定和特性研究

在成功筛选出1株能够降解微囊藻毒素(Microcystins,MCs)菌株的基础上,利用16S rDNA的PCR扩增和序列分析对其进行了分子鉴定。由BLAST序列比对及进化树分析可将该菌株定属至鞘氨醇单胞菌USTB-01(Sphingopyxissp.USTB-01),这是我国首次筛选出高效降解MCs的鞘氨醇单胞菌。在鞘氨醇单胞菌USTB-01降解MC-RR和MC-LR活性研究方面发现,温度30℃和pH7.0条件下该菌降解MCs的速率最大,日均降解MC-RR和MC-LR分别达到了23.4mg/L和7.9mg/L,在进一步去除水体中的MCs的基础和应用研究方面都具有非常重要的意义。     

微小小环藻对双酚A的富集与降解

对深圳福田红树林优势藻种微小小环藻(Cyclotella caspia)对BPA的生物富集与降解进行了研究.结果显示,微小小环藻对BPA具有较弱的富集能力,当BPA浓度为6.00、8.00、10.00 mg·L-1时,藻细胞在16d时分别达到最大富集量0.01、0.13、0.14 mg·g-1和最大生物富集系数(BCF)0.15%、1.18%、0.16%.微小小环藻对BPA亦具有一定的降解能力,浓度为6.00、8.00、10.00 mg·L-1的BPA在16d内分别有22.39%、36.44%和10.28%被藻降解,日平均生物降解量分别为0.08、0.18和0.06 mg·L-1.     

鄱阳湖微囊藻毒素时空分布格局及其与理化和生物因子的关系

本文分别于2012年4月(春季)、8月(夏季)、10月(秋季)和2013年1月(冬季)在鄱阳湖进行了4次野外调查和样品采集,采用超高效液相色谱-串联质谱仪(UPLC-MS/MS)对鄱阳湖水柱中常见的3种微囊藻毒素(MC-RR、MC-YR和MCLR)的质量浓度进行定量分析,探寻了微囊藻毒素(microcystins,MCs)质量浓度的时空变化规律及其与湖泊环境理化因子和生物因子之间的关系.结果表明,MC-RR是鄱阳湖MCs的主要毒素种类.胞内微囊藻毒素(intracellular MCs,IMCs)与微囊藻生物量(r=0.47,P0.01)、颤藻生物量(r=0.68,P0.01)、蓝藻生物量(r=0.56,P0.01)、Chl-a(r=0.28,P0.01)之间均具有显著的正相关关系,但与鱼腥藻生物量之间没有明显的相关关系(P0.05),这说明微囊藻和颤藻是鄱阳湖微囊藻毒素的主要产毒藻类.IMCs与水温(r=0.51,P0.01)、透明度(r=0.69,P0.01)、Fe(r=0.43,P0.01)和Zn(r=0.43,P0.01)之间均呈现显著的正相关关系,与TN(r=-0.44,P0.01)、TP(r=-0.29,P0.01)、NO-2-N(r=-0.28,P0.05)、NH+4-N(r=-0.33,P0.05)、Ca(r=-0.34,P0.01)和Mg(r=-0.35,P0.05)均呈现显著的负相关关系,但与pH、PO3-4-P、NO-3-N、电导率、高锰酸盐指数、Cu离子之间的相关关系不显著(P0.05),这说明光照强度(透明度反映)、氮、磷、水温是控制鄱阳湖微囊藻毒素产生的重要因子,金属离子Ca、Mg、Fe、Zn在一定程度上能影响微囊藻毒素的产生.鄱阳湖IMCs和胞外微囊藻毒素(extracellular MCs,EMCs)呈现相似的季节变化趋势,即夏季最高,其次为秋季,冬季和春季较低.鄱阳湖IMCs的空间分布整体上呈现东部湖汊群、松门山周围以及蚌湖和蚌湖进入鄱阳湖的入湖口较高,其它区域较低的趋势,而EMCs的高值区位于松门山周围和蚌湖及其入湖口.     

Removal of high concentrations of polycyclic aromatic hydrocarbons from contaminated soil by biodiesel

Solubilizing experiments were carried out to evaluate the ability of biodiesel to remove polycyclic aromatic hydrocarbons (PAHs) from highly contaminated manufactured gas plant (MGP) and PAHs spiked soils with hydroxypropyl-β-cyclodextrin (HPCD) and tween 80 as comparisons. Biodiesel displayed the highest solubilities of phenanthrene (420.7 mg·L⁻¹), pyrene (541.0 mg·L⁻¹), and benzo(a)pyrene (436.3 mg·L⁻¹). These corresponded to several fold increases relative to 10% HPCD and tween 80. Biodiesel showed a good efficiency for PAH removal from the spiked and MGP soils for both low molecular weight and high molecular weight PAHs at high concentrations. Biodiesel was the best agent for PAH removal from the spiked soils as compared with HPCD and tween 80; as over 77.9% of individual PAH were removed by biodiesel. Tween 80 also showed comparable capability with biodiesel for PAH solubilization at a concentration of 10% for the spiked soils. Biodiesel solubilized a wider range of PAHs as compared to HPCD and tween 80 for the MPG soils. At PAH concentrations of 229.6 and 996.9 mg·kg⁻¹, biodiesel showed obvious advantage over the 10% HPCD and tween 80, because it removed higher than 80% of total PAH. In this study, a significant difference between PAH removals from the spiked and field MGP soils was observed; PAH removals from the MGP soil by HPCD and tween 80 were much lower than those from the spiked soil. These results demonstrate that the potential for utilizing biodiesel for remediation of highly PAH-contaminated soil has been established.     

微囊藻毒素-LR对恶臭假单胞菌细胞活性和表面特性的影响

在恶臭假单胞菌(Pseudomonas putida)降解微囊藻毒素-LR(MC-LR)的体系中,研究了菌体细胞表面特性和活性的变化,考察了MC-LR对菌体的影响,对MC-LR的微生物毒性作用进行了初步的探索.结果表明,MC-LR可以使菌体细胞质膜通透性增加,并影响菌体离子代谢及可溶性蛋白等物质的分泌.当MC-LR质量浓度由0增大到2.0 mg·L-1时,能促进菌体分泌和释放可溶性糖及Na+、Cl-等离子.流式细胞实验检测发现,MC-LR加速了菌体细胞的死亡,且死亡率随着MC-LR质量浓度的增加而增大,2.5 mg·L-1MC-LR作用5 d,比未添加MC-LR的对照体系增加了近30%.扫描电镜观察表明,在MC-LR作用下,菌体细胞的微观形貌发生了改变,细胞褶皱加深,在2.5 mg·L-1MC-LR存在下培养5 d后,大部分细胞破裂,细胞内含物流出,细胞结构损坏严重.     

Trametes sp.LS-10C产漆酶发酵培养基优化及其漆酶对偶氮染料的脱色性能

为了提高菌株Trametes sp.LS-10C的漆酶产量并初步研究该酶对偶氮染料的脱色性能,本文通过单因素实验及均匀设计对菌株Trametes sp.LS-10C产漆酶培养基进行了优化,获得最优培养基配方为:葡萄糖14.4 g·L~(-1)、豆渣10.1 g·L~(-1)、麸皮100 g·L~(-1)、NH_4Cl_3g·L~(-1)、KH2PO41.4g·L~(-1)、CuSO_4’5H2O 1 g·L~(-1)、NaCl 1 g·L~(-1)、MgSO_40.8 g·L~(-1)、CaCl_20.5 g·L~(-1).菌株发酵至8 d时漆酶产量为595.15 U·m L~(-1),约是优化前的35.66倍.进一步研究表明,该漆酶-介体系统对酸性红GR、酸性蓝40和酸性铬蓝K等3种偶氮染料脱色10 h后脱色率分别高达96.86%、91.28%和86.31%.相比公开发表的文献,本研究所报道的Trametes sp.LS-10C利用优化培养基发酵产漆酶达到了较高水平,具有酶活力高和发酵时间短等特征,且该漆酶在处理偶氮染料废水脱色领域中具有进一步研发和应用价值.     

Impact of solids on biphasic biodegradation of phenanthrene in the presence of hydroxypropyl-β-cyclodextrin (HPCD)

The consequence of polycyclic aromatic hydrocarbons (PAHs) in the environment is of great concern. The hydrophobic properties of PAHs significantly impact phase distribution causing limited bioavailability. Enhanced biodegradation has been extensively carried out by surfactants and the redeployment effect was recognized. However, the quantitative relationship concerning the impact of solids was rarely reported. A batch of biphasic tests were carried out by introducing Mycobacterium vanbaalenii PYR-1 and hydroxypropyl-β-cyclodextrin (HPCD) into a mixture of phenanthrene solution and various glass beads (GB37-63, GB105-125, and GB350-500). The comparative results demonstrated that HPCD had little effect on microbial growth and was not degradable by bacterium. A model was proposed to describe the biodegradation process. The regression results indicated that the partition coefficient k _d (1.234, 0.726 and 0.448 L·g⁻¹) and the degradation rate k (0 mmol·L⁻¹: 0.055, 0.094, and 0.112; 20 mmol·L⁻¹: 0.126, 0.141, and 0.156; 40 mmol·L⁻¹: 0.141, 0.156 and 0.184 d⁻¹) were positively and negatively correlated with the calculated total surface area (TSA) of solids, respectively. Degradation enhanced in the presence of HPCD, and the enhancing factor f was calculated (20 mmol·L⁻¹: 15.16, 40.01, and 145.5; 40 mmol·L⁻¹: 13.29, 37.97, and 138.4), indicating that the impact of solids was significant for the enhancement of biodegradation.     

Relationships between loading rates and nitrogen removal effectiveness in subsurface flow constructed wetlands

Nitrogen removal of wetlands under 40 different inflow loadings were studied in the field during 15 months. The removal efficiency of four different sets of beds, namely the reed bed, the Zizania caduciflor bed, the mixing planting bed, and the control bed were studied. The outflow loading and total nitrogen (TN) removal rate of these beds under different inflow loadings and pollution loadings were investigated. The inflow loadings of 4 subsurface flow systems (SFS) ranged from 400 to 8000 mg·(m²·d)⁻¹, while outflow loadings were less than 7000 mg·(m²·d)⁻¹. The results showed that the inflow and outflow loading of TN removal rate in SFS presented an obvious linear relationship. The optical inflow loading to run the system was between 2000 to 4000 mg·(m²·d)⁻¹. Average removal rate was between 1062 and 2007 mg·(m²·d)⁻¹. SFS with plant had a better removal rate than the control. TN removal rates of the reed and Zizania caduciflora bed were 63% and 27% higher than the control bed, respectively. The results regarding the TN absorption of plants indicated that the absorption amount was very limited, less than 5% of the total removal. It proved that plants clearly increase TN removal rates by improving the water flow, and increasing the biomass, as well as activities of microorganisms around the roots. The research provided a perspective for understanding the TN removal mechanism and design for SFS. __________ Translated from Environmental Science, 2006, 27(2): 253–256 [译自: 环境科学]     

饱和带有机污染物的厌氧反硝化微生物降解

以卤代烃和芳香烃为代表,通过室内实验模拟饱和带厌氧反硝化环境,对地下饱和带中有机污染物的厌氧反硝化可生物降解性,降解机理,降解动力学方程等进行了研究。实验结果表明,芳香烃较卤代烃易被微生物降解,在有低学葡萄糖存在时有 机物的最大生物降解速率Ｃ６Ｈ６：Ｌｍａｘ＝１７ｍｇ／（ｍ＾２．ｄ）,Ｃ７Ｈ８：Ｌｍａｘ＝４６ｍｇ／（ｍ＾２．ｄ）,ＣＣｌ４;Ｌｍａｘ＝７ｍｇ／（ｍ＾２．ｄ）,ＣＨＣｌ３：Ｌｍａｘ＝６     

降解喹啉的微生物燃料电池的产电特性研究

通过构建双极室微生物燃料电池(Microbial fuel cell,MFC),对喹啉的降解及MFC的产电性能进行了研究.试验结果表明,当喹啉初始浓度为500 mg·L-1,葡萄糖与喹啉浓度之比为1:1,3:5,1:5时,MFC的最大输出电压分别为558 mV、469 mV、328 mV,运行周期分别为56.4 h、70h、82.5 h;最大功率密度分别为173 mW·m-2、122 mW·m-2、60 mW·m-2(按阳极截面积计算)或者35 W·m-3、24 W·m-3、12 W·m-3(按阳极室有效容积计算).MFC可实现对喹啉的高效降解,但葡萄糖的浓度对喹啉的降解速率有较大影响.当葡萄糖浓度分别为500 mg.L-1、300mg·L-1和100 mg·L-1时,使500 mg·L-1喹啉完全降解的时间分别为6 h、24 h和72 h.MFC闭路条件下对喹啉的降解速率高于开路厌氧条件下的喹啉降解速率约10%.MFC对喹啉的降解与产电速率之间存在差距,喹啉被快速降解至较低浓度(<5rag·L-1)后,MFC的产电性能才达到最优.MFC以用喹啉和葡萄糖作为混合燃料时,可以在实现高效降解喹啉的同时可稳定地向外输出电能,这为杂环芳烃类难降解有机物的高效低耗处理提供了新的途径.     

Effect of illumination on the hydrogen-production capability of anaerobic activated sludge

To investigate the influence of illumination on the fermentative hydrogen production system, the hydrogen production efficiencies of two kinds of anaerobic activated sludge (floc and granule) from an anaerobic baffled reactor were detected under visible light, dark and light-dark, respectively. The 10 mL floc sludge or granular sludge was respectively inoculated to 100 mL diluted molasses (chemical oxygen demand of 8000 mg·L⁻¹) in a 250 mL serum bottle, and cultured for 24 h at 37°C under different illumination conditions. The results showed that the floc was more sensitive to illumination than the granule. A hydrogen yield of 19.8 mL was obtained in the dark with a specific hydrogen production rate of 3.52 mol·kg⁻¹MLVSS·d⁻¹ (floc), which was the highest among the three illumination conditions. Under dark condition, the hydrogen yield of floc sludge reached the highest with the specific hydrogen production rate of 3.52 mol·kg⁻¹MLVSS·d⁻¹, and under light-dark, light, the specific hydrogen production rate was 3.11 and 2.21 mol·kg⁻¹MLVSS·d⁻¹, respectively. The results demonstrated that the illumination may affect the dehydrogenase activity of sludge as well as the activity of hydrogen-producing acetogens and then impact hydrogen production capacity.