活性污泥合成生物可降解塑料的研究进展

PHAs(polyhydroxyalkanoates,聚羟基脂肪酸酯 )是许多原核微生物在不平衡生长条件下合成的细胞内能量和碳源储藏性物质 ,作为完全可生物降解的热塑性聚酯而倍受关注。介绍了利用生物处理污水中活性污泥的混合碳源及微生物群体发酵合成PHAs的有关研究成果 ,包括微生物的种类、碳源及合成途径等 ,重点分析厌氧 好氧法活性污泥积累PHAs的量化关系 ,指出合理的工艺及适当的营养条件可以提高污泥中PHAs的积累量。此工作为污水处理资源化及污泥减量化、实现清洁生产提供了非常有潜力的研究方向。     

外加碳源的活性污泥合成生物降解塑料PHAs的试验

聚羟基脂肪酸酯(PHAs)是许多污泥微生物在不平衡生长条件下合成的细胞内能量和碳源储藏性物质,是一种新型的可完全生物降解的热塑性塑料.以污泥微生物作为PHAs的合成菌,采用乙酸钠、丙酸钠及丁酸钠作为单一碳源在好氧条件下合成得到占污泥干重5.58%、3.90%及4.98%的PHAs和在厌氧条件下合成得到占污泥干重12.32%、9.55%及11.35%的PHAs.实验表明,污泥微生物里广泛存在PHAs合成菌,厌氧条件比好氧条件的PHAs产率要高;最佳的实验条件下得到PHAs的量占污泥干重的12.32%,此时碳源转化率为30.65 %.此外,1HNMR谱图证明了同一污泥微生物用不同的碳源培养PHAs时其产物结构与组成均有变化.     

微生物利用食品废物合成聚羟基烷酸酯

为了降低聚羟基烷酸酯(PHAs)的生产成本,以麦芽、豆类、糖果、冰激凌、牛奶、芝麻油和食醋废物为底物,研究了产碱弧菌(Alcaligenes latus)、表皮匍萄球菌(Staphylococcus epidermidis)和活性污泥合成PHAs的工艺可行性和产物的物化性质.结果表明,通过两段式间歇补料发酵.3类微生物利用麦芽废物合成的聚羟基丁酸酯(PHB)产量最大,分别达细胞内含量的70.1%、16.0%和43.3%.A.latus适应食品废物开始细胞生长和PHAs合成的延迟短,在缺氮阶段合成PHB的产量、产率高.微好氧时,从芝麻油提取出的S.epidermidis可以合成分子量超过1×106的PHB.活性污泥可利用豆类废物合成聚羟基丁酸-羟基戊酸酯(PHBV)共聚物,其中羟基戊酸比例(HV%)占21%.多数食品废物适合合成具有不同物化性质的PHAs.PHAs产物的组分及物化性质受微生物种类、底物类型、发酵条件的影响.     

两类抗生素对EBPR系统的短期生物抑制作用实验研究

基于强化生物除磷(EBPR)系统,从除磷过程、挥发性脂肪酸(VFA)的消耗过程、聚羟基脂肪酸酯(PHAs)的合成与消耗过程、胞外聚合物(EPS)的总量变化过程及比呼吸速率(SOUR)的变化过程等多个角度出发,系统地研究了高浓度(10 mg·L~(-1))红霉素和土霉素对EBPR系统的短期冲击作用.结果表明,高浓度(10 mg·L~(-1))红霉素和土霉素在24 h之内均能对除磷过程产生明显的抑制作用,除磷效率分别下降至63.3%和61.2%.抗生素对VFA的消耗过程并无明显作用,而对PHAs的厌氧合成和好氧消耗过程影响较为显著.同时,高浓度(10 mg·L~(-1))抗生素对EPS中蛋白质(PN)合成量的抑制率在26.7%以上.SOUR变化过程的分析结果表明,抗生素对EBPR系统中微生物的呼吸作用抑制显著,在高浓度(10 mg·L~(-1))抗生素反应器中抑制率在16.0%以上.对比分析结果表明,相同浓度的土霉素比红霉素对EBPR系统的抑制更为显著,且EBPR系统的好氧过程比厌氧过程更容易受到抑制.     

A2/O污水处理工艺中基质转化机理研究

以实际污水培养驯化污泥的小试规模A2/O工艺为研究对象,对系统中基质的转化机理及硝态氮对基质转化的影响进行了批式试验研究.结果表明,在无硝态氮存在于厌氧环境的系统中,厌氧段消耗的COD有51%可被聚磷菌吸收并合成为聚羟基链烷酸(PHAs);缺氧和好氧条件下的比吸磷速率为3.87和6.54 mg/(g·h),利用单位PHAs的吸磷量(rP/PHA)分别为0.38和0.78而在有硝态氮存在于厌氧环境的系统中,厌氧段消耗的COD仅有30.8%可被聚磷菌吸收并合成PHAs,61.5%用于还原硝态氮;缺氧和好氧条件下的比吸磷速率为2.24和4.58 mg/(g·h),rP/PHA值分别为0.35和0.77.同时,在这2个系统中厌氧阶段释放的磷和消耗的COD成良好的线性关系.硝态氮存在于厌氧环境会降低聚磷菌的厌氧释磷速率和效率,使PHAs的合成量减少,从而降低聚磷菌的缺氧和好氧吸磷速率,但并不会影响其吸磷能力.     

溶解氧对好氧/延长闲置SBR除磷性能的影响

以合成废水为研究对象,乙酸钠为外加碳源,考察不同溶解氧(DO)浓度下好氧/延长闲置(O/EI)序批式反应器的除磷效果,并通过分析典型周期内磷元素及微生物体内各储能物质的变化,探究DO浓度对O/EI工艺除磷性能的影响机制.结果表明,低DO浓度(1 mg·L-1)条件下,O/EI系统具有良好的除磷效果,除磷率高达96%,单位污泥除磷量为5.02 mg·g-1;而当DO浓度较高(4 mg·L-1)时,反应器内磷的去除率降至50%,单位污泥除磷量仅2.81 mg·g-1.研究表明,在DO浓度为1 mg·L-1时,微生物能合成较多聚羟基脂肪酸酯(PHAs),糖原的合成及利用较少,系统好氧吸磷量远高于其他反应器,并在闲置期释放出更多聚磷酸盐.可见,DO可通过影响微生物体内PHAs和糖原的合成及转化,闲置期释磷,好氧前期释磷及好氧吸磷,进而影响系统的除磷性能.     

A2/O污水处理工艺中基质转化机理研究

以实际污水培养驯化污泥的小试规模A²/O工艺为研究对象,对系统中基质的转化机理及硝态氮对基质转化的影响进行了批式试验研究.结果表明,在无硝态氮存在于厌氧环境的系统中,厌氧段消耗的COD有51%可被聚磷菌吸收并合成为聚羟基链烷酸(PHAs);缺氧和好氧条件下的比吸磷速率为3.87和6.54 mg/(g·h),利用单位PHAs的吸磷量(r_P/PHA)分别为0.38和0.78.而在有硝态氮存在于厌氧环境的系统中,厌氧段消耗的COD仅有30.8%可被聚磷菌吸收并合成PHAs,61.5%用于还原硝态氮;缺氧和好氧条件下的比吸磷速率为2.24和4.58 mg/(g·h),r_P/PHA值分别为0.35和0.77.同时,在这2个系统中厌氧阶段释放的磷和消耗的COD成良好的线性关系.硝态氮存在于厌氧环境会降低聚磷菌的厌氧释磷速率和效率,使PHAs的合成量减少,从而降低聚磷菌的缺氧和好氧吸磷速率,但并不会影响其吸磷能力.     

正交定向调控VFAs奇偶数比率对合成PHAs的影响

为了定向调控挥发性脂肪酸(VFAs)中奇偶数碳比率,在合成聚羟基链烷酯(PHAs)过程中改变PHB和PHV的比率,从而改变可生物降解塑料的内在性能。以淘米水为单一发酵对象,将pH值、温度、发酵天数作为影响因素,通过L9(33)正交试验观察VFAs中奇偶数脂肪酸变化,和污泥中聚磷菌合成PHAs的变化规律。结果表明当温度为35℃,pH值为8,第7天时奇偶数碳比率约为1∶1;当温度为25℃,p H值为8,第9天时奇偶数碳比率约为1∶2;当温度为45℃,pH值为6,第7天时奇偶数碳比率约为1∶3。将这3种不同比率的组合等量投加至污泥中,PHAs含量有显著增加,其中发酵液奇偶数碳比率为1∶1时PHAs积累量最高,为80.69 mg/L。研究表明pH值对偶数脂肪酸影响最大,发酵天数对奇数脂肪酸影响最大。而且发酵液比化学碳源更有利于PHAs的合成,使厌氧阶段PHB的质量分数显著增加,好氧阶段PHV的质量分数显著增加。     

C/P对EBPR系统PAOs与GAOs竞争及PHAs代谢过程影响研究

以富含聚磷菌(PAOs)活性污泥为基础,基于FISH技术研究了SBR工艺不同C/P(25∶1、20∶1、15∶1和10∶1)对EBPR系统中功能菌变化特征与微生物胞内聚合物(PHAs)代谢过程的影响.结果表明,经过10 d运行处理,C/P分别为25∶1、20∶1和15∶1系统磷酸盐去除率88%,而C/P为10∶1系统磷酸盐的去除率为0%.FISH检测结果显示,随着C/P从25∶1下降到10∶1,EBPR系统中PAOs的含量相应从(76.42±1.24)%减少到(10.40±0.97)%,而聚糖菌(GAOs)则从(16.36±3.41)%增加到(34.25±2.59)%.在厌氧段,不同C/P条件下EBPR系统中PHB和PHV的合成动力学系数大小分别为K25∶1K20∶1K15∶1K10∶1和K15∶1K20∶1K25∶1K10∶1.随着C/P从25∶1下降到10∶1,合成PHB在PHAs中所占的比例从85%下降到24%,而PHV则从15%上升到76%;在好氧段,不同C/P系统消耗PHB和PHV的动力学系数大小均为K20∶1K25∶1K15∶1K10∶1,且C/P为25∶1、20∶1和15∶1时系统消耗主要成分是PHB(占PHAs 71%～75%),而C/P为10∶1时系统消耗主要成分是PHV(占PHAs 71%).由此表明,随着C/P的降低,EBPR系统内GAOs增加而PAOs减少,从而导致系统内PHB合成与消耗比例逐渐减少,而PHV合成与消耗比例逐渐增加.     

不同乙酸钠/甘油比对好氧/延长闲置SBR除磷性能的影响

以合成废水为研究对象,以甘油和生活污水中常见的乙酸钠作为碳源,建立了5个好氧/延长闲置序批式反应器(乙酸钠/甘油比分别为1∶0、4∶1、1∶1、1∶4和0∶1),考察了各反应器长期运行过程中的除磷效果,并通过分析典型周期内磷及微生物体内各储能物质的变化,初步探究不同乙酸钠/甘油比对除磷性能的影响机理.研究表明,当乙酸钠/甘油比由1∶0逐渐降至4∶1和1∶1时,平均除磷率由90.1%升至92.5%、97.3%.乙酸钠/甘油比继续降至1∶4及0∶1时,系统除磷率降至65.7%、53.4%.当乙酸钠/甘油比为1∶1时,聚磷菌体内合成大量聚羟基脂肪酸酯(PHAs)(2.55 mmol·g-1,以每g VSS积累的C(mmol)计,下同),为后续磷的吸收及聚磷合成提供更多的能量,而以甘油作为单一碳源时,PHAs合成量最少(0.82 mmol·g-1),糖原合成量最大(2.56 mmol·g-1,以每g VSS积累的C(mmol)计).     

基于CFD模拟的混合菌群合成PHA工艺筛选机制分析

利用数值模拟方法,建立了活性污泥混合菌群合成PHA工艺中ADD与ADF模式反应器的流体有限元分析模型,同时基于固液二相流体理论对ADD和ADF模式的沉淀过程进行分析,探讨了物理选择压的产生原理和作用机制。模拟获得了沉淀过程中不同停留时间反应器内不同沉降速率等级活性污泥的分布规律,以及沿反应器竖向方向的污泥体积分数分布规律。结果表明:ADD模式下活性污泥在沉淀过程中可形成较为稳定的沉降性差异分层界线,反应开始10~15 min内在高度为0.6 m处污泥分布体积分数为0.75左右,可对PHA合成能力强的菌群提供较强的筛选作用。固液二相流体场的模拟结果促进了对物理选择压筛选作用机制的认识,可为ADD模式反应器参数优化研究提供理论依据。     

Effects of sludge retention time, carbon and initial biomass concentrations on selection process: From activated sludge to polyhydroxyalkanoate accumulating cultures

Four sequence batch reactors (SBRs) fed by fermented sugar cane wastewater were continuously operated under the aerobic dynamic feeding (ADF) mode with different configurations of sludge retention time (SRT), carbon and initial biomass concentrations to enrich polyhydroxyalkanoate (PHA) accumulating mixed microbial cultures (MMCs) from municipal activated sludge. The stability of SBRs was investigated besides the enrichment performance. The microbial community structures of the enriched MMCs were analyzed using terminal restriction fragment length polymorphism (T-RFLP). The optimum operating conditions for the enrichment process were: SRT of 5 days, carbon concentration of 2.52 g COD/L and initial biomass concentration of 3.65 g/L. The best enrichment performance in terms of both operating stability and PHA storage ability of enriched cultures (with the maximum PHA content and PHA storage yield (Y_PHA/S) of 61.26% and 0.68 mg COD/mg COD, respectively) was achieved under this condition. Effects of the SRT, carbon concentration and initial biomass concentration on the PHA accumulating MMCs selection process were discussed respectively. A new model including the segmentation of the enrichment process and the effects of SRT on each phase was proposed.     

活性污泥微生物以乙酸为碳源合成聚羟基烷酸酯的研究

聚羟基烷酸酯(polyhydroxyalkanoate,PHA)是一种生物可降解塑料,利用活性污泥合成PHA具有操作方便的优点.本研究对长期驯化的厌氧-好氧活性污泥工艺在厌氧条件下及好氧活性污泥工艺在好氧条件下利用乙酸合成PHA进行了比较分析,发现好氧活性污泥可以合成较多的PHA.在此基础上,研究了好氧条件下乙酸加入次数、乙酸浓度及硝化抑制剂(丙烯基硫脲,ATU)对PHA合成的影响,结果表明,将乙酸分3次加入到反应器、乙酸浓度为2 925 mg/L及加入6.74 mg/L ATU时,可以使污泥挥发性悬浮固体物中PHA的含量达到56.3%.     

Bulking sludge for PHA production:Energy saving and comparative storage capacity with well-settled sludge

Two acetate-fed sequencing batch reactors (SBR) were operated under an aerobic dynamic feeding (ADF) model (SBR #2) and with anaerobic phase before aerobic phase (SBR #1) to select mixed cultures with a high polyhydroxyalkanoates (PHA) storage response. Although kinetic selection based on storage response should bring about a predominance of floc-formers, a bulking sludge with storage response comparable to well-settled sludge was steadily established. An anaerobic phase was introduced before the aerobic phase in the ADF model to improve the sludge settleability (SBR #1), however, due to the consequent increased feast/famine ratio, the performance of SBR #1, in terms of both the maximum PHB (polyhydroxybutyrate) cell content and ΔPHB, was lower than that of SBR #2. SBR #2 gradually reached a steady state while SBR #1 failed suddenly after 50 days of operation. The maximum specific substrate uptake rate and storage rate for the selected bulking sludge were 0.4 Cmol Ac/(Cmol X.hr) and 0.18 Cmol Ac/(Cmol PHB.hr), respectively, resulting a yield of 0.45 Cmol PHB/(Cmol Ac) in SBR #2 in the culture enrichment phase. A maximum PHB content of 53% of total suspended solids and PHB storage rate of 1.36 Cmol Ac/(Cmol PHB.hr) was achieved at 10.2 hr in batch accumulation tests under nitrogen starvation. The results indicated that it was feasible to utilize filamentous bacteria to accumulate PHA with a rate comparable to well-settled sludge. Furthermore, the lower dissolved oxygen demand of filamentous bacteria would save energy required for aeration in the culture enrichment stage.     

厌氧-好氧驯化活性污泥生物合成PHA的研究

厌氧-好氧处理有机污水的活性污泥中含有大量菌胶团菌,其形成与菌体胞内聚羟基烷酸酯(PHA)的积累有关,因此有可能利用污水中有机物和活性污泥中多种微生物合成PHA。笔者利用纺织工业废水厌氧-好氧驯化活性污泥,从中提取PHA。分别研究了厌氧过程和好氧过程中供氧量、碳源调节物浓度、培养时间等对菌胶团菌生长和胞内PHA积累的影响,测定了所得PHA的分子量、熔点和单体链节组成。      

缺氧条件下进水时间与溶解氧对底物贮存的影响

采用6个缺氧-好氧SBR反应器,考察了进水时间及溶解氧(DO)浓度对活性污泥系统中底物贮存的影响.缺氧进水条件下,进水时间的长短对底物贮存影响并不明显.进水时间由10min延长至60,90,120min时,各SBR系统内聚-β-羟基烷酸(PHA)贮存量依次小幅下降,最大差值为0.21mmolC/L, fPHB/HAc值在0.84~0.90范围内波动. 好氧曝气阶段控制低DO(0.5mg/L)运行比高DO(2.0mg/L)运行条件更有利于提高PHA的贮存量,90min进水时,高、低DO条件下PHA的平均合成量分别为3.1,5.0mmolC/L.而突然将进水时间90min缩短至10min,使得高、低DO系统中底物贮存量均增大,而高DO系统中底物贮存量的增长更为明显.     

Comparative study of polyhydroxyalkanoates production from acidified and anaerobically treated brewery wastewater using enriched mixed microbial culture

The production of polyhydroxyalkanoates(PHA) from wastewaters using microbial mixed cultures(MMC) has been attracting increased interest because of PHA's biodegradability characteristics. Production of PHA by an MMC enriched with PHA-accumulating bacteria was compared using anaerobically treated and acidified brewery wastewaters under various feeding strategies, namely pulse and batch feed addition. To obtain an enriched MMC, a sequencing batch reactor was inoculated with activated sludge fed with acetate and subjected to aerobic dynamic feeding. The enriched MMC was able to accumulate PHA up to 72.6% of cell dry weight(CDW) with pulse addition of acetate controlled by the dissolved oxygen(DO) concentration in the reactor. In a batch accumulation experiment with acetate,the PHA content achieved(28.5% CDW) was less than that of the pulse feeding strategy with the same amount of acetate(～2000 mg C/L). Using anaerobically treated and acidified brewery wastewater fed in pulses, the maximum PHA accumulated by the enriched MMC was similar for both wastewaters(45% CDW), in spite of the higher volatile fatty acid concentration in acidified brewery wastewater. The pulse feed addition controlled by the DO concentration was difficult to implement for wastewater as compared to acetate because the difference in DO concentration between substrate availability and depletion was low. For the batch addition of acidified wastewater, a slightly lower PHA content(39%CDW) was obtained. These results show that both brewery wastewaters can be utilized for PHA production with a similar maximum PHA storage capacity.     

不同碳源种类对好氧颗粒污泥合成PHA的影响

影响好氧颗粒污泥稳定性的因素众多,其中碳源种类的不同会造成好氧颗粒污泥合成聚羟基烷酸酯(PHA)的不同,进而影响其稳定性能.采用混合碳源驯化培养的好氧颗粒污泥进行试验.考察了厌氧条件下,乙酸钠等八种碳源对好氧颗粒污泥合成PHA的影响.结果表明,颗粒污泥对乙酸钠和蔗糖具有较好的转化能力,合成PHA的量分别为102.19mgCOD/g·VSS和70.58mgCOD/g·VSS,显著高于其他碳源的PHA合成量(5~26mg/g×VSS).故以蔗糖和乙酸钠作碳源均有利于颗粒污泥稳定性能的维持,而当以甲醇作碳源时好氧颗粒污泥的贮存能力最差.     

溶解氧对活性污泥胞内贮存物和除磷性能的影响

分别以乙酸钠和葡萄糖为碳源,通过间歇实验,考察不同溶解氧(DO)浓度对厌氧-缺氧-好氧序批式反应器中活性污泥胞内贮存物聚羟基烷酸酯(PHA)、糖原和系统除磷性能的影响。实验表明:DO对以乙酸钠为碳源培养的胞内贮存物PHA和糖原影响较大,并影响污泥的除磷性能;对以葡萄糖为碳源培养的胞内物质影响较小,可能是因为细胞内存在其他类型的胞内贮存物。     

Effects of phosphorus and nitrogen limitation on PHA production in activated sludge

The e ects of phosphorus and nitrogen limitation on polyhydroxyalkanoate (PHA) production and accumulation by activated sludge biomass with acetate as a carbon source were investigated. Pre-selected influent carbon-phosphorus (C:P, W/W) of 100, 160, 250, 500 and 750, and carbon-nitrogen (C:N, W/W) of 20, 60, 100, 125 and 180 were applied in the phosphorus limitation experiments and the nitrogen limitation experiments, respectively. The maximum PHA accumulation up to 59% of the cell dry weight with a PHA productivity of 1.61 mg PHA/mg COD consumed was observed at the C:N 125 in the nitrogen limitation experiment. This value was much higher than that obtained in previous studies with a normal substrate feeding. The study showed that activated sludge biomass would produce more polyhydroxybutyrate than polyhydroxyvalerate under the stress of nutrient limitation, especially under phosphorus limitation conditions. The experimental result also indicated that both phosphorus and nitrogen limitation may cause sludge bulking.