Optimal biocompatible solvent design for a two-stage extractive fermentation process with cell recycling

In this study, crisp and fuzzy multiple-goal optimization approaches are respectively introduced to design an optimal biocompatible solvent to a two-stage extractive fermentation with cell recycling for ethanol production. When designing a biocompatible solvent for the extractive fermentation process, many issues, such as extractive efficiency, conversion, amount of solvent utilized and so on, have to be considered. An interactive multiple-goal design procedure is introduced to determine a trade-off result in order to satisfy such contradicted goals. Both approaches could be iterated to solve the interactive multiple-goal design problem in order to yield a trade-off result. However, the crisp optimization design is a tedious task that requires the designer to provide various pairs of the upper bounds for the design problem to obtain the corresponding solution. The fuzzy optimization approach is able to be trade-off several goals simultaneously and to yield the overall satisfactory grade for the product/process design problem.     

Effects of growth with ethanol on fermentation and membrane fluidity of Saccharomyces cerevisiae

Saccharomyces cerevisiae HSc was grown with ethanol at concentrations up to 10% (v/v). The immediate effects of additions of externally added ethanol on CO₂ production and O₂ consumption of washed organisms were studied by stopped-flow membrane inlet quadrupole mass spectrometry. Fermentative activities of organisms grown with ethanol (0–5% v/v) showed similar sensitivities to inhibition by ethanol, whereas those grown with 10% (v/v) ethanol had become protected and were markedly less sensitive. The fluidity of subcellular membrane fractions was measured by determination of the temperature dependence of the rotational order parameter of the spin label 5-doxyl stearic acid (free radical) by electron spin resonance. Mitochondria prepared from yeasts grown with 0, 7 and 9% (v/v) ethanol showed similar overall fluidity, although differences in temperature-dependent behaviour indicate altered lipid composition or lateral phase separations. On the other hand the microsomal fraction from organisms grown with 9% ethanol showed a remarkable increase in fluidity. These data suggest that the protective effects of growth with ethanol near the limit of tolerance on fermentative activities may arise from altered plasma membrane fluidity properties.     

工业发酵的生物学属性对发酵过程最优控制的制约与对策

从工业发酵过程计算机控制的历史和现状出发，探讨了工业发酵过程的生物学属性对发酵过程最优化控制的制约，以及解决这个难题的对策。在论证了工业发酵具有明显的生物学属性的基础上，阐明了发酵的生物学原理及其在发酵过程最优化控制中应用的重要性。     

Mathematical modelling of the alcoholic fermentation of glucose by Zymomonas mobilis mobilis

The kinetics of alcoholic fermentation of a strain of Zymomonas mobilis, isolated from sugarcane juice, has been studied with the objective of determining the constansts of a non-structured mathematical model that represents the fermentation process. Assays in batch and in continuous culture have been carried out with different initial concentrations of glucose. The final concentrations of glucose, ethanol and biomass were determined. The following kinetic parameters were obtained: μ_max, 0·5 h^?1; K_s, 4·64 g dm^?3; P_max, 106 g dm^?3; Y_x/s, 0·0265 g g^?1; m, 1·4 g g^?1 h^?1; α, 17·38 g g^?1; β, 0·69 g g^?1 h^?1.     

EM发酵饲料中主要微生物类群的动态变化

对EM发酵饲料中乳酸杆菌、酵母菌、大肠菌群、腐败菌等微生物类群的动态变化进行了研究。结果表明 :饲料经EM发酵后 ,乳酸杆菌大量增加 ,发酵第4天达到最大值(9.5×108CFU·g-1 饲料) ,大肠菌群与腐败菌被明显抑制。并认为细菌总数与酵母菌数量不能作为评价EM发酵饲料质量好坏的指标。     

黑曲霉葡萄糖氧化酶在毕赤酵母中的表达及其高效发酵研究

葡萄糖氧化酶是一类能催化β-D-葡萄糖生成葡萄糖酸-δ-内酯的酶类,在食品、药品以及生物传感器等领域具有广泛的应用前景。从黑曲霉中克隆出葡萄糖氧化酶编码序列,在毕赤酵母中实现了组成型和诱导型表达并分析了其酶学性质和发酵条件。结果表明:黑曲霉葡萄糖氧化酶的编码基因长度为1 818 bp,编码605个氨基酸,具有21个氨基酸组成的信号肽。毕赤酵母组成型表达葡萄糖氧化酶的发酵上清液中葡萄糖氧化酶的酶活可达1.2 U/m L,利用镍离子亲和层析纯化后得到了分子质量约为100 kDa的葡萄糖氧化酶。利用糖苷内切酶H(Endo H)对葡萄糖氧化酶去糖基化后,其分子质量约为70 kDa,与推测的分子质量相近。对黑曲霉葡萄糖氧化酶的酶学性质研究表明,该酶的最适反应pH 6.0,最适反应温度35℃,比活力60.9 U/mg。为了进一步提高葡萄糖氧化酶在毕赤酵母中的表达量,构建了诱导型表达的毕赤酵母菌株并通过G418抗生素筛选获得了高效表达菌株。对该菌株进行高密度发酵(湿菌体200 g/L),甲醇添加量为2.5%,发酵至第7天时葡萄糖氧化酶发酵量可达133 U/m L,蛋白表达量约1.9 g/L。     

细菌纤维素合成过程中的影响因素

研究了接种量、pH值、培养液深度、培养液表面积和发酵时间等因素对形成细菌纤维素膜的产量、厚度及比表面积的影响。结果表明:接种量在7%、初始pH值在5～6范围内细菌纤维素的产量最大;厚度主要取决于发酵时间和初始pH值;培养液表面积对单位面积细菌纤维素产量有比较大的影响;膜的比表面积与发酵时间的长短无关。     

玉米秸秆生物法制取酒精的中间试验

建立了玉米秸秆采用蒸汽爆破预处理、纤维素酶水解和戊糖己糖同步发酵技术制取酒精的中间试验装置。玉米秸秆在1.6～2.0 MPa条件下蒸汽爆破预处理,在提高玉米秸秆对纤维素酶可及度的同时,玉米秸秆中纤维素、木聚糖和木质素损失分别为4.08%、40.02%和9.91%。里氏木霉以10%的原料制备纤维素酶,并用于降解剩余的90%的原料,滤纸酶活力和纤维素酶水解得率分别为2.27 FPIU/mL和71.3%。初始还原物浓度为43.65 g/L的水解糖液经树干毕赤酵母发酵16 h,还原物利用率和酒精得率分别为87.17%和0.43 g/g(酒精/消耗的糖)。     

高效液相色谱法测定柠檬酸的含量

建立了一种准确对发酵液中柠檬酸含量测定的仪器方法,在检测范围6-16 mg/mL之间成线性,R=0.999 7,RSD为±0.8%-1.2%(n=6),最低检测浓度为0.02 mg/mL.此方法简单、灵敏、重现性好,避免了由于化学转化所引起的测定误差,以及发酵液中色素的影响.     

提高发酵过程乙醇分离效率的新型反应器

设计了一种新型的带侧臂循环的气升式反应器,该反应器可以有效地把真空闪蒸发酵和汽提发酵2项技术结合起来。实验结果表明,与单独的汽提和闪蒸过程相比,利用此反应器进行同步汽提闪蒸操作,大大提高了乙醇的移出效率,从而进一步提高了发酵强度。