溶氧及pH对地衣芽孢杆菌合成聚γ-谷氨酸的影响

在3.7L发酵罐中研究了溶氧、pH和甘油流加对地衣芽孢杆菌分批发酵生产聚γ-谷氨酸(γ-PGA)的影响.结果表明,当葡萄糖浓度为27.9g/L且通气量控制在4L/min时,搅拌转速达到300r/min即可满足细胞生长和聚γ-谷氨酸合成对溶解氧的需求.不同pH控制方式对聚γ-谷氨酸分批发酵的影响有较大差异.不控制pH时,细胞干重和聚γ-谷氨酸产量比控制pH为5.5的发酵分别低26%和94%.研究了将pH控制在4.0、4.5、5.0、5.5、6.0和6.5的聚γ-谷氨酸分批发酵过程,发现在pH5.5时聚γ-谷氨酸总产量最高.以溶氧水平作为甘油代谢指针来控制甘油限制性流加既可维持一定菌体生长,又不会发生发酵液中残余甘油及有害代谢产物阻遏作用.菌体关于甘油的表观的率、聚γ-谷氨酸的平均比生产速率较没有采用甘油限制性流加时都有所提高.图4表1参7     

氮源及其添加模式对钝齿棒杆菌JDN28-75合成L-精氨酸的影响

氮源是微生物过量合成L-精氨酸的重要营养因子之一,不同氮源对钝齿棒杆菌JDN28-75合成L-精氨酸的影响研究结果表明,硫酸铵为合适的氮源.不同初始硫酸铵浓度对JDN28-75产L-精氨酸的影响研究结果表明,氮源浓度过高或不足,都会使最终L-精氨酸产量有所降低.低浓度的硫酸铵虽然有利于菌体生长,但对L-精氨酸的合成明显不利,同时糖酸转化率也较低;而高浓度的硫酸铵尽管不利于细胞的生长且造成发酵结束时残糖含量过高,却有利于细胞合成L-精氨酸且实际耗糖的糖酸转化率维持在一个较高的水平.初始硫酸铵浓度为60 g/L时,对JDN28-75菌体的生长有明显的抑制作用,最终发酵液中剩余的硫酸铵也较多(大于30 g/L),但高浓度的硫酸铵是L-精氨酸合成所必需的.在上述研究结果的基础上,确定了初始硫酸铵浓度为20 g/L条件下的补氮策略,比较了4种不同的硫酸铵补加模式对产L-精氨酸的影响,结果表明,在总的硫酸铵浓度相同的情况下,采取分批、低浓度添加氮源的方式既可以有效解除发酵前期高浓度硫酸铵对菌体生长的抑制作用,又可以有效维持发酵中后期体系中菌体合成L-精氨酸所需的较高比例的氮源.最后,在5 L全自动发酵罐中采用20 g/L的初始硫酸铵浓度,连续流加25%的氨水来控制发酵体系pH及补加氮源,L-精氨酸的产量可以达到31.7 g/L,较对照组的产酸量(26.0 g/L)提高了21.9%.图4表2参11     

废电池中重金属溶出特性及对海洋生物的毒性效应(Dissolved Characteristics of Heavy Metal in Waste Battery and Toxic Effect on Marine Organisms)

针对渔用废电池被大量丢弃在海洋中的现象,分别开展了废电池中主要重金属离子溶出特性试验和废电池浸出液对不同海洋生物急性毒性效应的研究.试验结果显示,在盐度为20的40L海水中自然浸泡状态下(45节电池),松下一号锌锰废电池溶出液中铅、镉、汞溶出浓度不断增加,但溶出速率较慢.单节电池在第60d,铅、镉和汞溶出总量分别为2.08μg、0.52μg和0.60μg,溶出率分别为0.004%、0.018%和1.263%;第210d铅、镉和汞溶出总量分别为28.76μg、6.38μg和1.02μg,溶出率分别为0.057%、0.224%和2.147%.一节废电池中铅、镉和汞总量在1L海水中全部溶出后浓度分别可达到50445μg·L-1、2850μg·L-1和47.5μg·L-1,分别是我国渔业水质标准(GB11607-89)的1009倍、570倍和95倍.废电池浸出液对不同受试生物的急性毒性试验结果表明,当废电池浸出液混合浓度中铅、镉和汞浓度分别为3.39μg·L-1、0.64μg·L-1和0.76μg·L-1时(45节电池40L海水浸泡60d),对黑鲷、脊尾白虾和缢蛏的96h半致死浓度值分别为溶出液混合浓度的5.13%、4.87%和6.71%,废电池浸出液中各重金属离子对海洋生物毒性具有非常强的协同作用.在鱼、虾、贝三类受试生物中,贝类对废电池溶出液毒性的耐受能力最强,鱼类次之,虾类最弱.     

低浓度溶解氧下猪粪固体有机物水解产酸研究

本文以猪粪为发酵原料,通过批式实验研究不同溶解氧(DO)浓度(0—0.26 mg.L-1)、发酵时间(3—12 d)和挥发性固体(VS)浓度(11.14—111.35 g.L-1)对猪粪固体水解酸化过程的影响,确定了低DO浓度下固体有机物优化水解产酸工艺条件:中温35℃,初始VS浓度37.11 g.L-1,初期的DO浓度0.1—0.26 mg.L-1.在此条件下,发酵时间3 d,DO浓度下降到0.10 mg.L-1以下.猪粪发酵液pH值由7.45±0.10降低到5.86±0.17,VS降解率(25.67±1.20)%,挥发性脂肪酸(VFA)中乙酸(3895±91)mg.L-1、丙酸(2313±82)mg.L-1、正丁酸(1361±17)mg.L-1、正戊酸(540±11)mg.L-1.优化条件下的猪粪水解酸化液进行厌氧产甲烷发酵,发酵10 d内产气停止,低溶解氧水解酸化和厌氧产甲烷发酵累计时间仅为13 d,甲烷体积分数(69.5±0.2)%,VS产气率为(0.282±0.011)L CH.4g-1VS.研究结果表明,适当延长发酵时间能够增加VFA中乙酸的含量,初始VS浓度差异对发酵液VFA浓度和VS降解率的影响较显著,低溶解氧能够促进猪粪固体有机物水解过程.     

温度和发酵时间对厨余垃圾发酵产乳酸及其光学异构体的影响

通过间歇实验,探讨了温度和发酵时间对非灭菌的厨余垃圾发酵产乳酸及其光学异构体的影响.结果表明,控制温度在35℃时,发酵液中乳酸浓度远高于45℃和55℃,发酵120h后,总乳酸的产率和产量最大,分别达到0.59gL-1d-1和0.62gg(VS)-1,以有机碳表示的乳酸占发酵液中总有机碳的78%;控制发酵时间小于120h,L-乳酸在35℃、45℃和55℃三个温度条件下都是主要的异构体形式;35℃和55℃时,发酵72h后,随着发酵时间的延长,L-乳酸浓度在总乳酸中的比例呈下降趋势;45℃时,L-乳酸浓度随着发酵时间的延长逐渐增加,L-乳酸在总乳酸中的比例在发酵144h后达到97%.图4表1参21     

流加发酵及添加L-半胱氨酸对产朊假丝酵母高密度培养合成谷胱甘肽的影响

通过7L罐考察了不同葡萄糖流加速率以及添加L-半胱氨酸对产朊假丝酵母(Candida utilis)WSH02-08高密度发酵生产谷胱甘肽(GSH)的影响.结果表明,在恒定速度(5.5g L-1h-1)流加葡萄糖30h的基础上,发酵45h后细胞干重最高达到73g L-1,此时向发酵罐中一次性添加L-半胱氨酸40mmol L-1,最终GSH产量和胞内GSH含量分别达到了1458mg L-1和2.26%.图4表1参11     

初期通气和震荡培养提高高浓度乙醇发酵的乙醇浓度和产率

研究了氧气和震荡条件对酿酒酵母高浓度乙醇发酵的影响.结果表明,震荡是提高发酵液乙醇浓度和产率的最重要因素.与静止培养相比,在不通气情况下震荡培养使乙醇浓度提高了69%(从75.8 g L-1提高到128.1 g L-1),在通气条件下乙醇浓度提高了68.7%(从85.2 g L-1提高到to 143.8 g L-1).在最优条件下,两次补料,经54 h发酵,发酵液中乙醇浓度达到143.8 g L-1,乙醇产率与理论产率的比值为0.471 g/g(即92.20%).经分析,通气和震荡条件提高了发酵液中酿酒酵母的生物量和细胞活力.图5表1参12     

顺丁烯二酸异构酶工程菌发酵条件优化

为提高重组大肠杆菌中顺丁烯二酸异构酶表达量,通过正交试验设计,对工程菌的生长条件和目的蛋白可溶表达条件进行优化.采用250 mL三角瓶中装有50 mL(Amp 100 mg L-1)的培养基,分别研究培养基中葡萄糖、蛋白胨、酵母浸粉的浓度,培养基pH值以及摇床转速、装液量、接种量等对蛋白可溶表达量的影响.确定顺丁烯二酸异构酶工程菌最优化培养基为:蛋白胨20 g L-1、酵母浸粉2.5 g L-1、K2HPO4·3H2O 3.0 g L-1、KH2PO4 1.5 g L-1、NaCl 6 g L-1、MgSO43 g L-1,培养基pH调至6.5.确定顺丁烯二酸异构酶工程菌可溶性表达最优条件为:37℃下培养至D600 nm值为1.0时,添加终浓度为0.05 mmol L-1的IPTG进行诱导,诱导温度37℃,摇床转速220 r min-1,装液量20%,接种量5%,诱导时长为6 h.利用BioFlo 415发酵罐以最优化的培养基和发酵条件对该工程菌进行了3批发酵实验,与摇瓶实验相比,顺丁烯二酸异构酶的表达量提高了近1.5倍,单位发酵液的酶活力由46 U mL-1发酵液提高到78 U mL-1发酵液.以上数据为顺丁烯二酸异构酶重组工程菌的中试发酵奠定了基础.图7表2参17     

混合培养微生物利用甘油补料发酵生产乙醇研究

采用浸麻芽孢杆菌和红曲菌990691用甘油混合发酵生产乙醇.结果表明,分批发酵中高浓度的甘油对乙醇发酵有着较强的抑制作用,分批发酵最佳甘油浓度为0.217 mol L-1.在分批发酵的基础上补料发酵,考察了不同甘油浓度的补料液和不同补料时间对乙醇发酵的影响.最终确定乙醇补料发酵较优的工艺条件为:反应器1 L,装液量700 mL红曲发酵液,甘油初始浓度为0.217 mol L-1,以补料方式每隔60 h分5次补加0.217 mol L-1甘油浓度的红曲发酵液,每次补加100 mL,发酵培养360 h.当乙醇最高浓度达0.221 mol L-1,乙醇总产率0.628 mmol h-1,乙醇/甘油转化率达87%(mol mol-1).与分批发酵相比,补料发酵很大程度解除了高浓度甘油的抑制作用,有效地利用了甘油,提高了乙醇的产量,且乙醇产率较为稳定.     

Streptomyces sp. WJS-825产谷氨酰胺转胺酶发酵条件优化及中试

应用五因子二次正交旋转回归试验设计,建立了微生物谷氨酰胺转胺酶 (TG)发酵生产过程中以酶活力和菌体细胞生长量作为目标函数的数学模型,并以此模型对链霉菌 (Streptomycessp. )WJS-825菌株发酵生产TG的培养条件进行优化,确定了影响TG生产的主要因子及其最适取值为多价胨 2. 1%、可溶性淀粉 1. 5%、初始pH 7. 0及培养温度 30℃.以该优化工艺条件进行了 5L发酵罐小试和 200L发酵规模的中试生产.结果表明,在中试发酵生产中使用以豆饼粉部分替代多价胨的经济性发酵优化培养基,以及发酵过程中在线监控pH、溶氧系数等多项发酵调控参数,并分段控制pH、温度、通气量和搅拌转速以及进行适时的流加补充碳源,该菌株生长繁殖能力强、产酶效果好,TG活性达 3. 2u/mL,而且连续重复的中试发酵生产试验的TG产量均稳定在 3. 2u/mL以上. 图 2表 3参 15     

Experimental study of nitrite accumulation in predenitrification biological nitrogen removal process

The effect of dissolved oxygen (DO) concentration on nitrite accumulation was investigated in a pilot-scale pre-denitrification process at room temperature for 100 days. In the first 10 days, due to the instability of the system, the DO concentration fluctuated between 1.0 and 2.0 mg/L. In the next 14 days, the DO concentration was kept at 0.5 mg/L and nitrite accumulation occurred, with the average nitrite accumulation rate at 91%. From the 25th day, the DO concentration was increased to 2.0 mg/L to destroy the nitrite accumulation, but nitrite accumulation rate was still as high as 90%. From the 38th day the nitrite accumulation rate decreased to 15%–30% linearly. From the 50th day, DO concentration was decreased to 0.5 mg/L to resume nitrite accumulation. Until the 83rd day the nitrite accumulation rate began to increase to 80%. Dissolved oxygen was the main cause of nitrite accumulation, taking into account other factors such as pH, free ammonia concentration, temperature, and sludge retention time. Because of the different affinity for oxygen between nitrite oxidizing bacteria and ammonia oxidizing bacteria when DO concentration was kept at 0.5 mg/L, nitrite accumulation occurred.     

Impact of dissolved oxygen on the production of nitrous oxide in biological aerated filters

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and microelectrode technology were employed to evaluate the Nitrous oxide (N₂O) production in biological aerated filters (BAFs) under varied dissolved oxygen (DO) concentrations during treating wastewater under laboratory scale. The average yield of gasous N₂O showed more than 4-fold increase when the DO levels were reduced from 6.0 to 2.0 mg?L^–1, indicating that low DO may drive N₂O generation. PCR-DGGE results revealed that Nitratifractor salsuginis were dominant and may be responsible for N₂O emission from the BAFs system. While at a low DO concentration (2.0 mg?L^–1), Flavobacterium urocaniciphilum might play a role. When DO concentration was the limiting factor (reduced from 6.0 to 2.0 mg?L^–1) for nitrification, it reduced NO ₂ ^- -N oxidation as well as the total nitrification. The data from this study contribute to explain how N₂O production changes in response to DO concentration, and may be helpful for reduction of N₂O through regulation of DO levels.

Open image in new window

     

Facile fabrication of dolomite-doped biochar/bentonite for effective removal of phosphate from complex wastewaters

● Dolomite-doped biochar/bentonite was synthesized for phosphate removal. ● DO/BB exhibited a high phosphate adsorption capacity in complex water environments. ● PVC membrane incorporated with DO/BB can capture low concentration phosphate. ● Electrostatic interaction, complexation and precipitation are main mechanisms. The removal of phosphate from wastewater using traditional biological or precipitation methods is a huge challenge. The use of high-performance adsorbents has been shown to address this problem. In this study, a novel composite adsorbent, composed of dolomite-doped biochar and bentonite (DO/BB), was first synthesized via co-pyrolysis. The combination of initial phosphate concentration of 100 mg/L and 1.6 g/L of DO/BB exhibited a high phosphate-adsorption capacity of 62 mg/g with a removal efficiency of 99.8%. It was also stable in complex water environments with various levels of solution pH, coexisting anions, high salinity, and humic acid. With this new composite, the phosphate concentration of the actual domestic sewage decreased from 9 mg/L to less than 1 mg/L, and the total nitrogen and chemical oxygen demand also decreased effectively. Further, the cross-flow treatment using a PVC membrane loaded with DO/BB (PVC-DO/BB), decreased the phosphate concentration from 1 to 0.08 mg/L, suggesting outstanding separation of phosphate pollutants via a combination of adsorption and separation. In addition, the removal of phosphate by the PVC-DO/BB membrane using NaOH solution as an eluent was almost 90% after 5 cycles. The kinetic, isotherm and XPS analysis before and after adsorption suggested that adsorption via a combination of electrostatic interaction, complexation and precipitation contributed to the excellent separation by the as-obtained membranes.     

溶解氧及活性污泥浓度对同步硝化反硝化的影响

文章研究溶解氧及活性污泥浓度对同步硝化反硝化效率的影响。研究结果表明：在一定DO范围内，随着反应器内溶解氧浓度的降低，总氮去除率呈上升趋势，即好氧反硝化效率随溶解氧浓度的降低而提高；在一定MLSS范围内，反应器内混合液污泥浓度越高，出水总氮越低，反硝化现象越明显。     

Micro-analysis of nitrogen transport and conversion inside activated sludge flocs using microelectrodes

To investigate the nitrogen transport and conversion inside activated sludge flocs, micro-profiles of O₂, NH₄⁺, NO₂^–, NO₃^–, and pH were measured under different operating conditions. The flocs were obtained from a laboratory-scale sequencing batch reactor. Nitrification, as observed from interfacial ammonium and nitrate fluxes, was higher at pH 8.5, than at pH 6.5 and 7.5. At pH 8.5, heterotrophic bacteria used less oxygen than nitrifying bacteria, whereas at lower pH heterotrophic activity dominated. When the ratio of C to N was decreased from 20 to 10, the ammonium uptake increased. When dissolved oxygen (DO) concentration in the bulk liquid was decreased from 4 to 2 mg·L^-1, nitrification decreased, and only 25% of the DO influx into the flocs was used for nitrification. This study indicated that nitrifying bacteria became more competitive at a higher DO concentration, a higher pH value (approximately 8.5) and a lower C/N.     

低溶解氧条件下生物脱氮研究中的新现象

活性污泥法中 ,曝气主要起供氧和扰动混合的作用 ,曝气提供的氧被微生物用来氧化有机物并合成细胞 .反应器中的溶解氧 (DO)浓度是重要的运行参数 ,曝气池中DO偏低 ,好氧微生物不能正常生长和代谢 ;DO过高 ,不仅能耗增加 ,而且细菌的活力也会降低 .一般要求曝气池中DO不低于 2mgL-1的水平 ,但在实践中常常会出现曝气强度过高的情况 .因而有必要通过有效手段将DO控制在适当的水平 ,既不影响微生物的正常生长和有机物的去除 ,同时又避免过多能耗 .传统观点认为 ,低DO条件会促进丝状菌生长 ,破坏污泥絮体的沉降性能 ;使胞外多聚物的产生量…     

SBR短程硝化的实现、维持及稳定性

以模拟生活污水为研究对象,控制SBR反应器内pH值在7.5～8.5的条件下,实现了短程硝化生物脱氮工艺,NO2--N/NOx--N的比率始终维持在90%以上,同时发现pH值和DO浓度变化特征曲线在短程硝化过程中具有良好的重现性。另外,保持DO浓度在0.5～1.0mg/L,硝化时间为5.5h,可较好的维持短程硝化生物脱氮过程,且经过1个月的运行硝化类型没有发生改变,亚硝酸盐积累率仍保持在90%以上。在此基础上,研究了系统对COD和NH4+-N浓度的抗冲击负荷能力。结果表明,该系统具有较强的抗冲击能力。     

Performance of completely autotrophic nitrogen removal over nitrite process under different aeration modes and dissolved oxygen

In this study, three sequential batch biofilm reactors (SBBRs) were operated for 155 days to evaluate the performance of completely autotrophic nitrogen removal over nitrite (CANON) process under different aeration modes and dissolved oxygen (DO). Synthetic wastewater with 160-mg NH4 ⁺-N/L was fed into the reactors. In the continuously-aerated reactor, the efficiency of the ammonium nitrogen conversion and total nitrogen (TN) removal reached 80% and 70%, respectively, with DO between 0.8–1.0 mg/L. Whereas in the intermittently-aerated reactor, at the aeration/non-aeration ratio of 1.0, ammonium was always under the detection limit and 86% of TN was removed with DO between 2.0–2.5 mg/L during the aeration time. Results show that CANON could be achieved in both continuous and intermittent aeration pattern. However, to achieve the same nitrogen removal efficiency, the DO needed in the intermittently-aerated sequential batch biofilm reactor (SBBR) during the aeration period was higher than that in the continuously-aerated SBBR. In addition, the DO in the CANON system should be adjusted to the aeration mode, and low DO was not a prerequisite to CANON process.     

Nitrification and denitrification in biological activated carbon filter for treating high ammonia source water

Since the ammonia in the effluent of the traditional water purification process could not meet the supply demand, the advanced treatment of a high concentration of NH₄ ⁺-N micro-polluted source water by biological activated carbon filter (BACF) was tested. The filter was operated in the downflow manner and the results showed that the removing rate of NH₄ ⁺-N was related to the influent concentration of NH₄ ⁺-N. Its removing rate could be higher than 95% when influent concentration was under 1.0 mg/L. It could also decrease with the increasing influent concentration when the NH₄ ⁺-N concentration was in the range from 1.5 to 4.9 mg/L and the dissolved oxygen (DO) in the influent was under 10 mg/L, and the minimum removing rate could be 30%. The key factor of restricting nitrification in BACF was the influent DO. When the influent NH₄ ⁺-N concentration was high, the DO in water was almost depleted entirely by the nitrifying and hetetrophic bacteria in the depth of 0.4 m filter and the filter layer was divided into aerobic and anoxic zones. The nitrification and degradation of organic matters existed in the aerobic zone, while the denitrification occurred in the anoxic zone. Due to the limited carbon source, the denitrification could not be carried out properly, which led to the accumulation of the denitrification intermediates such as NO₂ ^?. In addition to the denitrification bacteria, the nitrification and the heterotrophic bacteria existed in the anoxic zone.