Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited (C/N ratio is 56/8.8 mmol/L) medium. Several sets of shaking flask experiments were conducted. The results showed that 2 g/L glucose feeding on the first day of the culture (24 h after the inoculation) simulated both fungal biomass growth and enzyme production. The manganese peroxidase (MnP) activity was 2.5 times greater than that produced in cultures without glucose feeding. Furthermore, the glucose feeding mode in fed-batch culture was also investigated. Compared to cultures with glucose feeding every 48 h, cultures with glucose feeding of 1.5 g/L (final concentration) every 24 h produced more enzymes. The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture, respectively, and the enzyme was kept stable for 4 days with an activity of over 200 U/L. Translated from Acta Scientise Circumstantiae, 2007, 27(3): 363–368 [译自: 环境科学学报]     

Enhanced production of poly(3-hydroxybutyrate) by filamentation-suppressed recombinantEscherichia coli in a defined medium

Fed-batch cultures of recombinantEscherichia coli strains were carried out for the production of poly(3-hydroxybutyric acid) (PHB) in a chemically defined medium. TheE. coli strains used were XL1-Blue, harboring pSYL105, a stable high-copy number plasmid containing theAlcaligenes eutrophus polyhydroxyalkanoate (PHA) genes, and XL1-Blue, harboring pSYL107, which is pSYL105 containing theE. coli ftsZ gene to suppress filamentation. With XL1-Blue(pSYL105) the final cell mass and PHB concentration obtained in 62 h were 102 and 22.5 g/L, respectively. Fed-batch culture of XL1-Blue(pSYL107) under identical conditions resulted in a final cell mass and PHB concentration of 127.5 and 48.2 g/L, respectively. The PHB contents obtained with XL1-Blue(pSYL105) and XL1-Blue(pSYL107) were 22.1 and 37.8%, respectively. Therefore, PHB was more efficiently produced in a defined medium by employing filamentation-suppressed recombinantE. coli.     

Highly efficient conversion of sugarcane bagasse pretreated with liquid hot water into ethanol at high solid loading

Liquid hot water (LHW), an environmental-friendly physico-chemical treatment, was applied to pretreat the sugarcane bagasse (SCB). Tween80, a non-ionic surfactant, was used to enhance the enzymatic hydrolysis of the pretreated SCB. It found that 0.125 mL Tween80 /g dry matter could make the maximum increase (33.2%) of the glycan conversion of the LHW-pretreated SCB. A self-designed laboratory facility with a plate-and-frame impeller was applied to conduct batch hydrolysis, fed-batch hydrolysis, and the process of high-temperature (50°C) fed-batch hydrolysis following low-temperature (30°C) simultaneous saccharification and fermentation (SSF) which was adopted to overcome the incompatible optimum temperature of saccharification and fermentation in the SSF process. After hydrolyzing LHW-pretreated SCB for 120 h with commercial cellulase, the total sugar concentration and glycan conversion obtained from fed-batch hydrolysis were 91.6 g/L and 68.3%, respectively, which were 9.7 g/L and 7.3% higher than those obtained from batch hydrolysis. With Saccharomyces cerevisiae Y2034 fermenting under the non-sterile condition, the ethanol production and theoretical yield obtained from the process of SSF after fed-batch hydrolysis were 55.4 g/L and 88.3% for 72h, respectively, which were 15.5 g/L and 24.7% higher than those from separate fed-batch hydrolysis and fermentation. The result of this work was superior to the reported results obtained from the LHW-pretreated SCB.     

4-氯酚存在对生物强化系统降解2,4-二氯酚的影响研究

采用生物强化技术降解废水中的难降解有机物 2 ,4 二氯酚 (简称 2 ,4 DCP) ,研究了不同浓度的 4 氯酚 (简称 4 MCP)存在对 2 ,4 DCP降解的影响 ,并通过半连续流实验研究了 4 MCP长期存在下 ,强化系统中 2 ,4 DCP和 4 MCP降解速率的变化趋势 .结果表明 ,第 1次半连续流实验中 ,4 MCP浓度为 5、10、2 0及 30mg L时 ,都会对强化系统中 2 ,4 DCP的降解速率产生一定的抑制作用 ,而且抑制作用随着 4 MCP浓度的增加而增强 .随着半连续流实验次数的增加 ,4 MCP对 2 ,4 DCP降解的抑制作用减弱甚至消失 ,共基质时 2 ,4 DCP的降解速率反而比其单基质半连续流运行时快 .4 MCP与 2 ,4 DCP共基质存在时 ,2 ,4 DCP优先被微生物利用 ,此后 4 MCP才被降解 ,表现出两阶段降解的现象 .     

采用序半连续反应器进行硝化和反硝化工艺的数学模拟

研究了生物废水处理应用于序半连续式反应器(SFBR)工艺中的微生物生长、硝化和反硝化动力学。基于活性污泥1号模型(ASMNo. 1)中的原理和结论,导出了在有氧阶段和缺氧阶段微生物生长速率、铵、硝酸盐、易生物降解基质等的反应速率及它们的物料平衡方程。在实验的基础上,对模型中的各参数采用了合适的方法进行参数估值,即:龙格库塔法解常微分方程组和黄金分割法搜索最小误差,该模型的最小误差ε≈4 799;得出了应用于本工艺中数学模型中的各动力学参数和化学计量系数,如KS 39. 997,KNO 0 .397,KNH 1 997,KOA 0 .404,KOH 0. 297, μA 0 .0026,μH 0. 207,YA 0 .24,YH. 0 6。     

基于DO-stat流加培养控制的L-异亮氨酸发酵条件优化

L-异亮氨酸(L-lie)产生菌代谢流的前期研究报道认为,较低的溶氧浓度有利于L-异亮氨酸的积累.为进一步提高L-异亮氨酸的产量,采用DO-stat流加培养控制方法,分别研究了溶氧浓度20%、30%、35%和40%下乳糖发酵短杆菌发酵液中葡萄糖浓度的变化以及L-异亮氨酸的合成情况.结果表明,当发酵罐中溶氧浓度控制在20...     

Temperature and feed strategy effects on sulfate and organic matter removal in an AnSBB

The objective of this work was to analyze the interaction effects between temperature, feed strategy and COD/[SO(4)(2-)] levels, maintaining the same ratio, on sulfate and organic matter removal efficiency from a synthetic wastewater. This work is thus a continuation of Archilha et al. (2010) who studied the effect of feed strategy at 30 °C using different COD/[SO(4)(2-)] ratios and levels. A 3.7-L anaerobic sequencing batch reactor with recirculation of the liquid phase and which contained immobilized biomass on polyurethane foam (AnSBBR) was used to treat 2.0 L synthetic wastewater in 8 h cycles. The temperatures of 15, 22.5 and 30 °C with two feed strategies were assessed: (a) batch and (b) batch followed by fed-batch. In strategy (a) the reactor was fed in 10 min with 2 L wastewater containing sulfate and carbon sources. In strategy (b) 1.2 L wastewater (containing only the sulfate source) was fed during the first 10 min of the cycle and the remaining 0.8 L (containing only the carbon source) in 240 min. Based on COD/[SO(4)(2-)] = 1 and on the organic matter (0.5 and 1.5 gCOD/L) and sulfate (0.5 and 1.5 gSO(4)(2-)/L) concentrations, the sulfate and organic matter loading rates applied were 1.5 and 4.5 g/L.d, i.e., same COD/[SO(4)(2-)] ratio (=1) but different levels (1.5/1.5 and 4.5/4.5 gCOD/gSO(4)(2-)). When reactor feed was 1.5 gCOD/L.d and 1.5 gSO(4)(2-)/L.d, gradual feeding (strategy b) showed to favor sulfate and organic matter removal in the investigated temperature range, indicating improved utilization of the electron donor for sulfate reduction. Sulfate removal efficiencies were 87.9; 86.3 and 84.4%, and organic matter removal efficiencies 95.2; 86.5 and 80.8% at operation temperatures of 30; 22.5 and 15 °C, respectively. On the other hand, when feeding was 4.5 gCOD/L.d and 4.5 gSO(4)(2-)/L.d, gradual feeding did not favor sulfate removal, indicating that gradual feeding of the electron donor did not improve sulfate reduction.     

微生物浓度对SFBR工艺去除营养物的影响

采用序半连续式反应器(SequentialFed-batchReactor,简称SFBR,下同。)就不同微生物初始浓度对人工配置废水营养物去除效率的影响进行了研究。探讨了微生物初始浓度对COD、氮化合物(NH4-N和NO3-N)除去和微生物生长的影响。结果表明,当微生物初始浓度为140.4g/L时,COD有最高除去率(69.5%)。当微生物初始浓度≥46.8g/L,总氮除去率可达100%。菌体初始浓度对废水中营养物的去除有利。过高浓度的微生物生长,受营养物和供氧的限制。用一组涉及多个微生物反应的动力学模型,分析了微生物初始浓度对同时去除碳、氮化合物效率和微生物生长的影响。     

白腐真菌木质素降解酶的反应器发酵

为了获得更高的白腐真菌木质素降解酶的产量和相应的控制策略,应用5L搅拌罐生物反应器对氮限制下(C/N=56/2.2)P.chrysosporium产木质素降解酶进行了放大研究.结果表明,在分批发酵试验中,锰过氧化物酶(MnP)和漆酶(Lac)分别在培养第6d和第7d达到峰值,其酶活随时间的变化规律与摇瓶试验基本相同;而采用氮限制液体培养基进行补料没有获得更高的酶活,因此,可以得出采用发酵液体培养基作为补料液不利于白腐真菌持续产酶.另外,在分批发酵和分批补料发酵过程中,均发现体系pH值变化与白腐真菌生长和次生代谢产酶具有较好的相关性,当白腐真菌进入次生代谢阶段开始产酶时,体系pH值开始下降,随着发酵后期酶活的降低,pH值下降的幅度也逐渐变小,当体系酶活接近为0.0U/L时,pH值基本不再变化.因此,在实际发酵过程中,可以依据体系pH的变化间接了解白腐真菌的生长和产酶情况,而分批补料策略还有待于进一步研究.     

Cell-recycle fed-batch production of a highly unsaturated polyhydroxyalkanoate from 1,3-butanediol byPseudomonas sp. A33

The recently isolatedPseudomonas sp. A33 was investigated for the production of a highly unsaturated polyhydroxyalkanoate (PHA) containing various alkyl and alkenyl pendent groups from 1,3-butanediol in a cell-recycle fed-batch production mode. The monomer composition and degree of unsaturation in PHA were dependent on the environmental conditions. The production temperature markedly influenced the content, composition, and degree of unsaturation of PHA. As the production temperature decreased from 30 to 10°C, the degree of unsaturation and content of PHA were increased, while the mole percentage of 3-hydroxybutyrate (3HB) was decreased. These temperature effects on the composition of PHA imply that the production can be used as a control variable for the biosynthesis of a highly unsaturated PHA and for the specific regulation of the composition of PHA. The biosynthetic pathway for a highly unsaturated PHA which is based on de novo fatty acid biosynthetic pathway is proposed. For the enhanced production of this functional PHA, a high cell density was achieved by cell-recycle continuous culture at 30°C, and then a large amount of PHA was accumulated at 15°C by fed-batch addition of the feeding solution containing excess 1,3-butanediol. The structures of monomer constituents of polymer were confirmed by gas chromatography—mass spectrometric analysis of trimethylsiyl derivatives of 3-hydroxyalkanoic acids methyl esters.