pH及流加葡萄糖对酵母分批发酵生产谷胱甘肽的影响

在5 L的发酵罐中研究了pH及流加葡萄糖对酵母分批发酵生产谷胱甘肽(GSH)的影响。实验考察了不同浓度的流加葡萄糖和不同的恒pH值的分批发酵对于酵母生产GSH产量的变化。实验结果表明,当pH值控制为5.0,流加葡萄糖流速为5g.L-1.h-1,连续流加30 h,可使GSH产量最高,与之前未流加葡萄糖和控制pH相比,其产量提高了6倍。     

采用最小二乘支持向量机的青霉素发酵过程建模研究

生化过程通常是严重非线性和时变的复杂动态系统,而且重要过程参数缺少在线测量仪表,对其建立机理模型往往非常耗时和困难。采用最小二乘支持向量机(LS_SVM)并以Pensim仿真平台为例对青霉素发酵这一典型生化过程进行建模研究。给出了LS_SVM参数的调整策略和分析结果,建立了青霉素产物浓度、菌体浓度和底物浓度等重要过程变量的在线预报模型。仿真结果表明用LS_SVM建立的在线预报模型拟合误差小,推广性能好,可以作为发酵过程的进一步控制和优化的参考依据。     

分装过程中尘粒变化的影响因素探讨

通过对在线粒子实时监测系统采集的尘粒变化数据进行分析,探讨影响分装过程中尘粒变化的因素。对同批次分装时四个监测点和不同批次分装时同一监测点尘粒变化的数据进行分析,结果表明,在分装的前期准备、后期清场以及排除故障期间,粒子数变化明显。可见洁净间人员的数量及人员的行为是影响分装过程中尘粒变化的主要因素。     

On-line self-checking of replication consistency for autonomic computing

In this paper we are concerned with the live verification of the consistency of a replicated system, an issue that has not been addressed by the research community so far. We consider the problem of how to enable the system to detect automatically and in production whether the invariants defining the correctness of object replication are violated. This feature could greatly improve the dependability of distributed applications and is necessary for constructing self-managing and self-healing replicated systems. We focus on systems that enforce strongly consistent replication: all replicas of each object must be kept “continuously” in-sync. This replication strategy is appropriate for application domains where correctness guarantees in spite of failures are more important than performance and scalability. We present the design and implementation of a replicated web service capable of self-checking whether all replicas are indeed kept in sync. This check occurs on-line, transparently to clients. We also discuss the performance cost of self-checking in our prototype. Alberto Bartoli is Associate Professor of Computer Engineering at the University of Trieste, Italy. He took a degree in Electrical Engineering in 1989 and a doctorate in Computer Engineering in 1994, both at the University of Pisa, Italy. His research interests are in the area of reliability and fault-tolerance in distributed systems. Giovanni Masarin took a degree in Electronic Engineering in 2004, at the University of Trieste, Italy. He is currently involved in product development at RadioTrevisan, a company specialized in the production of lawful interception equipments.     

Over-expression of recombinant human interferon-gamma in high cell density fermentation of Escherichia coli

Human interferon-gamma (hIFN-gamma) was expressed in Escherichia coli BL21(DE3) under the control of the T7 promoter. Glucose was used as the sole source of carbon and energy with simple exponential feeding rate in fed-batch process. Cell density of recombinant E. coli was reached to 100 g dry wt l(-1) under both constant (0.12 h(-1)) and variable (0.12-0.52 h(-1)) specific growth rates. In the variable specific growth rate fed-batch process, plasmid stability and specific yield of rhIFN-gamma were greater than constant specific growth rate fed-batch process. The final specific yield and overall productivity of rhIFN-gamma were 0.35 +/- 0.02 g rhIFN-gamma g(-1) dry cell wt and 0.9 +/- 0.05 g rhIFN-gamma l(-1) h(-1) in the variable specific growth rate fed-batch process, respectively.     

Poly(3-hydroxybutyrate) synthesis in fed-batch culture of Ralstonia eutropha with phosphate limitation under different glucose concentrations

Poly(3-hydroxybutyrate) (PHB) was produced by fed-batch cultures of Ralstonia eutropha with phosphate limitation under different glucose concentrations. When glucose was kept at 2.5 g l^–1, cell growth and PHB synthesis were limited due to the shortage of carbon source but a higher PHB content occurred in the cell-growth stage. This shows that a low glucose concentration is favorable for PHB accumulation in R. eutropha. PHB obtained with glucose at 9 g l^–1 is 1.6 times that obtained with 40 g l^–1. When glucose was in the range of 9 to 40 g l^–1, PHB concentration and productivity decreased significantly with the increase of glucose concentration. The highest PHB productivity was obtained with glucose at 9 g l^–1.     

Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris

The gene coding for agglutinin from Galanthus nivalis (GNA) was expressed in, and secreted by, the methylotrophic yeast, Pichia pastoris. Transformants of P. pastoris were selected and a process to produce and purify gram quantities of recombinant GNA was developed. GNA was secreted at approximately 80 mg l^–1 at the 200 l scale and was purified to 95% homogeneity using hydrophobic interaction chromatography. The recombinant protein was similar to the protein synthesised in plant with respect to structure and biological activity.     

Optimization of growth conditions of the postharvest biocontrol agent Candida sake CPA-1 in a lab-scale fermenter

AIM: To maximize the growth (expressed as number of viable cells per millilitre) of the postharvest biocontrol agent Candida sake CPA(-1) at laboratory scale conditions. METHODS AND RESULTS: Growth conditons (aeration, agitation speed and inoculum size) were studied in batch conditions in a 5 l fermenter using molasses and urea as growth medium. Consumption of sugars and urea were analysed. Fed-batch studies were also carried out. Glucose and fructose were consumed during the exponential growth phase and were depleted after 18 h of growth. On the contrary, C. sake cells assimilated sucrose during the stationary phase. There was not growth improvement when fed-batch technology was used. Addition of an extra amount of glucose or molasses after 18 h of growth did not contribute to increase final population. CONCLUSIONS: Maximum growth (about 8 x 10(8 )CFU ml(-1)) was obtained at batch fermentation after 30 h growth at 400 rev min(-1), 150 l h(-1) of air and initial concentration of 106 CFU ml(-1). SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained in this study are an approach for further upscaling of C. sake production.     

Optimization of fed-batch fermentation for xylitol production by Candida tropicalis

Xylitol, a functional sweetener, was produced from xylose by biological conversion using Candida tropicalis ATCC 13803. Based on a two-substrate fermentation using glucose for cell growth and xylose for xylitol production, fed-batch fermentations were undertaken to increase the final xylitol concentration. The effects of xylose and xylitol on xylitol production rate were studied to determine the optimum concentrations for fed-batch fermentation. Xylose concentration in the medium (100 g l⁻¹) and less than 200 g l⁻¹ total xylose plus xylitol concentration were determined as optimum for maximum xylitol production rate and xylitol yield. Increasing the concentrations of xylose and xylitol decreased the rate and yield of xylitol production and the specific cell growth rate, probably because of an increase in osmotic stress that would interfere with xylose transport, xylitol flux to secretion to cell metabolism. The feeding rate of xylose solution during the fed-batch mode of operation was determined by using the mass balance equations and kinetic parameters involved in the equations in order to increase final xylitol concentration without affecting xylitol and productivity. The optimized fed-batch fermentation resulted in 187 g l⁻¹ xylitol concentration, 0.75 g xylitol g xylose⁻¹ xylitol yield and 3.9 g xylitol l⁻¹ h⁻¹ volumetric productivity. Journal of Industrial Microbiology & Biotechnology (2002) 29, 16–19 doi:10.1038/sj.jim.7000257 Received 15 October 2001/ Accepted in revised form 30 March 2002     

Fed-batch cultivation of animal cells using different medium design concepts and feeding strategies

In animal cell cultivation, cell density and product concentration are often low due to the accumulation of toxic end-products such as ammonia and lactate and/or the depletion of essential nutrients. A hybridoma cell line (CRL-1606) was cultivated in T-flasks using a newly devised medium feeding strategy. The goals were to decrease ammonia and lactate formation by the design of an initial medium which would provide a starting environment to achieve optimal cell growth. This was followed by using a stoichiometric equation governing animal cell growth and then designing a supplemental medium for feeding strategy used to control the nutritional environment. The relationship between the stoichiometric demands for glutamine and nonessential amino acids was also studied. Through stoichiometric feeding, nutrient concentrations were controlled reasonably well. Consequently, the specific production rate of lactate was decreased by fourfold compared with conventional fed-batch culture and by 26-fold compared with conventional batch culture. The specific production rate of ammonia was decreased by tenfold compared with conventional fed-batch culture and by 50-fold compared with conventional batch culture. Most importantly, total cell density and monoclonal antibody concentration were increased by five- and tenfold respectively, compared with conventional batch culture. (c) 1994 John Wiley & Sons, Inc.