Artificial neural network based experimental design procedure for enhancing fermentation development

Conventional experimental design techniques are available to assist in the optimization of fermentation processes, but due to the nonlinearities in the bioprocess, they are limited in their effectiveness. This problem is further complicated with recombinant systems as a result of the additional complexities of the process. This article describes a general strategy using artificial neural networks as an alternative approach to fermentation process development laboratory are presented for the neural network based procedures. (c) 1994 John Wiley & Sons, Inc.     

Bioprocess monitoring by marker gene analysis

The optimization and the scale up of industrial fermentation processes require an efficient and possibly comprehensive analysis of the physiology of the production system throughout the process development. Furthermore, to ensure a good quality control of established bioprocesses, on-line analysis techniques for the determination of marker gene expression are of interest to monitor the productivity and the safety of bioprocesses. A prerequisite for such analyses is the knowledge of genes, the expression of which is critical either for the productivity or for the performance of the bioprocess. This work reviews marker genes that are specific indicators for stress- and nutrient-limitation conditions or for the physiological status of the bacterial production hosts Bacillus subtilis, Bacillus licheniformis and Escherichia coli. The suitability of existing gene expression analysis techniques for bioprocess monitoring is discussed. Analytical approaches that enable a robust and sensitive determination of selected marker mRNAs or proteins are presented.     

Hierarchical control of biotechnological complex systems

Application of hierarchical control for solution of some nonlinear optimization problems in fermentation systems, with the use of microprocessors, is described. Results presented show some advantages of the method of hierarchical control in comparison to some traditional one-level optimization methods. The advantage of the hierarchical control method is in less demand on memory and computing power of the control computer as the standard methods.In solving this problem two control schemes are used. In the first control scheme the method of objective coordination is used. In the second one the prediction method of coordination is used, with the aim to minimize selected disturbances in state and control variables.     

An integrated microprocessor-based fermenter control system

An integrated microprocessor-based fermenter controller was developed in 1980 for an operational environment at Cetus Corp. The main goals in the design and construction of the system were (1) to facilitate scale-up; (2) to provide flexibility and high performance for optimizing fermentation processes; and (3) to be cost-effective for 15 in-house systems. It was also developed to work in conjunction with a laboratory minicomputer for on-line optimization experiments. The controller controls temperature, agitation, dissolved oxygen, pH, and foam throughout each fermentation run without manual intervention. The feedback control parameters have been optimized to provide very accurate control over a wide range of setpoint conditions and under rapidly changing metabolic conditions such as induced during an Escherichia coli batch run. The controller has also been configured to monitor, display, and record each of the controlled variables; support the interactive operator console; and communicate with the laboratory computer. In over 4 years of operation, these systems have met the design goals and have proven to be very reliable. The controller is described, its operational performance presented, and a typical fermentation run delineated.     

New methods of oxygen monitoring in the gas phase of cultivation processes

Summary The principles of two new techniques for monitoring oxygen in the gas phase of fermentation processes are presented. In addition, the application of both methods on real fermentation processes is given to show their potential in comparsion with well established methods.     

Numerical aspects of optimal control of penicillin production

Since their discovery, fermentation processes have gone along not only with the industrial beverages production and breweries, but since the times of Alexander Fleming, they have become a crucial part of the health care due to antibiotics production. However, complicated dynamics and strong nonlinearities cause that the production with the use of linear control methods achieves only suboptimal yields. From the variety of nonlinear approaches, gradient method has proved the ability to handle these issues—nevertheless, its potential in the field of fermentation processes has not been revealed completely. This paper describes constant vaporization control strategy based on a double-input optimization approach with a successful reduction to a single-input optimization task. To accomplish this, model structure used in the previous work is modified so that it corresponds with the new optimization strategy. Furthermore, choice of search step is explored and various alternatives are evaluated and compared.     

Some practical aspects of computer applications in a fermentor hall

The use of computers in a fermentation pilot plant is described from a practical point of view. The aim is not the application of a computer to a single special process but a general application of computers to prepare and present data for the following analysis of fermentation processes. The hardware is normally bought from computer manufacturers, but some additional installations are useful. Application software has to be developed by the biotechnologist, therefore the software structure is the most important part of the computer application. Data storage is divided into three parts: short-time memory, long-time storage, and a medium memory mainly to be used in process analyses and process control. Four types of programs are used: main schedule tasks, low-priority on-line tasks, sense-switch routine, and different off-line programs. A table of all programs is presented, the main schedule task is described in detail to demonstrate the software structure.     

Mikrocomputer-unterstützte prozeßkontrolle in der fermentationstechnik

Nowadays computer control is an effective and well established tool in optimizing fermentation processes. Based on recent development in microelectronics, especially in the field of personal computers, cost effective applications of this technology are now possible, even on laboratory- and pilot scale. In this paper the application of a personal computer (CBM 3032), controlling various types of fermentations, is described. In more detail the availability of physiologically relevant sensors, especially the respiration and the dynamic calorimetry, are discussed. Finally some examples of practical realisations are presented.     

Automatic computer control of fermentation processes

The application of an optimization algorithm to fermentation processes is described, which is based on information received by means of cascading the input quantities with pseudo random binary signals. The results in optimization of temperature, pH-value as well as productivity are presented and discussed.     

Integrated system for bioprocess monitoring and diagnosis using a local area network

Bioprocess monitoring and data analysis utilizing a local area network (LAN) is described in which an integrated computer environment provides for real-time monitoring from several remote personal computers with easy evaluation of the current process status and providing a common utilization of fermentation data. The computer network also enhances the decision-making process in the management of the production plant. Bioprocess control utilizing a LAN environment is expected to promote better utilization of fermentation data accumulated through repeated operations and to realize advanced control of fermentation processes. © Rapid Science Ltd. 1998     

Feedback optimization of fed-batch fermentation

The problem of feedback optimization of the feed rate for fed-batch fermentation processes is formulated in the framework of singular control theory and switching hypersurfaces. Using four differential balance equations that describe a general class of fedbatch processes and a general objective function to be minimized, it is shown that under certain restrictions the feedback optimization of the feed rate can be realized as a nonlinear function of the state variables, such as the concentrations of cell mass, substrate and product, and the fermentor volume. The restrictions on the initial conditions, the fermentation kinetics and the objective function, that are needed for realization of the feedback optimization, are provided. Fed-batch fermentation models of lysine and alcohol are used to construct switching curves and to illustrate the feedback optimization of the feed flow rates.     

Pattern analysis techniques to process fermentation curves: application to discrimination of enological alcoholic fermentations

In fermentation processes, kinetic curves are generally aimed at control purposes. However, these curves could also contain information about inherent features of the product (such as origin, quality, etc.). This article presents several pattern analysis techniques used to classify fermentation curves. An application to alcoholic fermentation is presented as an illustration: it aims at retrieving the origin of a must from its fermentation curve. The fermentation kinetics of five vineyard musts, harvested over 9 years on the same parcels, were recorded. From these curves two sets of variables were generated: The first (p(1)) gathers all the kinetic curve points. The second (p(2)) contains a restrained number of variables, generated by the expert knowledge of the enologist. The set p(2) was processed by two very different techniques: a linear one (factorial discriminant analysis) and a nonlinear one (artificial neural networks). The set p(1) was processed by a new chemometric technique, the discriminant partial least-squares regression. For all the sets and the techniques used the selection of variables was studied. The interest in the latter is largely demonstrated both by theoretical and practical discussions. The discrimination results (up to 94% of good predictions) enhance the interest of the on-line measurements and their use in such pattern analysis tools.     

试验设计和优化及其在发酵培养基优化中的应用

在发酵工业中,发酵培养基的优化对发酵水平的提高起着举足轻重的作用。而在寻求最佳发酵培养基的过程中,试验设计及统计优化发挥着重要作用。对近年来常用的试验设计及优化方法进行了综述,内容包括单因素轮换法、析因设计、均匀设计、响应面设计、人工神经网络和遗传算法等多种试验设计和优化方法,并对其进行分析和比较。     

灵芝胞外多糖分批发酵非结构动力学模型

在２Ｌ搅拌发酵罐上提出了描述了灵芝胞外多糖分批发酵过程中菌球生长、底物消耗和胞外多糖形成的非结构动力学模型。首先研究了灵芝分批发酵特性,结果表明该发酵过程属菌体生长和产物形成相偶联型。然后在总结文献的基础上,运用动力学模型,经过非线性回归,得到了模型中的参数值。通过计算机模拟,证明模型预测值与实际实验值具有良好的拟合性。     

Application of rRNA-targeted oligonucleotide probes in biotechnology

Ribosomal RNA-targeted oligonucleotide probes have become valuable tools for the detection of microorganisms involved in important biotechnological processes. Microorganisms which are of major importance for processes such as wastewater treatment, microbial leaching or methane production can be detected and quantified in situ within a complex microbial community. For certain processes, such as nitrification or biological phosphate removal, new microorganisms have become the focus of interest and have led to an improved understanding of these bioremediation techniques. Hybridization techniques have become fast and reliable alternatives to conventional cultivation techniques in the food industry as a control method for starter cultures for fermentation processes or product control. Recent analytical tools such as flow cytometry and digital image processing have improved the efficiency of these techniques. This review is intended to present a summary of methodological aspects of rRNA-based hybridization techniques and their application in biotechnology.     

Optimization of batch fermentor sterilization

This article presents a calculation procedure useful for the optimization and scale up of batch sterilization cycles in large-scale fermentors. This technique determines the sterilization temperature and hold-time necessary to minimize nutrient damage in a specific fermentor. The method can also be used for "scaledown" experiments to eliminate sterilization conditions as a scale up parameter. A method for the systematic evaluation of different sterilization conditions on product yield is also presented. This procedure is useful in determining if scale up of sterilization conditions is important for a given process. The validity of the techniques presented are supported by data showing significant yield improvements in a 1.2 x 10(5) L antibiotic fermentation.     

Monitoring of recombinant protein production using bioluminescence in a semiautomated fermentation process

On-line optimization of fermentation processes can be greatly aided by the availability of information on the physiological state of the cell. The goal of our "BioLux" research project was to design a recombinant cell capable of intracellular monitoring of product synthesis and to use it as part of an automated fermentation system. A recombinant plasmid was constructed containing an inducible promoter that controls the gene coding for a model protein and the genes necessary for bioluminescence. The cells were cultured in microfermenters equipped with an on-line turbidity sensor and a specially designed on-line light sensor capable of continuous measurement of bioluminescence. Initial studies were done under simple culture conditions, and a linear correlation between luminescence and protein production was obtained. Such specially designed recombinant bioluminescent cells can potentially be applied for model-based inference of intracellular product formation, as well as for optimization and control of recombinant fermentation processes.     

Unstructured model for growth of mycelial pellets in submerged cultures

An unstructured model is presented to describe growth of mycelial pellets in submerged cultures. This model integrates growth kinetics at the scale of the hyphae with the physical mechanisms of mass-transfer processes at the scale of the pellets and the fermentor. The main elements of the model are biomass, substrate, and oxygen balances for the liquid phase and the pellets. The possible occurrence of oxygen limitation in the pellets is introduced in analogy with catalyst theories by means of an effectiveness factor. To simulate the growth of pellets the model is transferred into a computer program. The model is tested by means of fermentation experiments in a bubble column. Results of the growth experiments compare favorably with the outcome of computer simulations.     

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。     

Computer control of the penicillin fermentation using the filtration probe in conjunction with a structured process model

A structured model for the penicillin fermentation is presented. This model includes three different cell types: (1) hyphae tips, (2) penicillin-producing cells, and (3) degenerated, metabolically inactive cells. Cell degeneration has been described previously as a gradual loss of cytoplasmic material by endogenous metabolism. The rate at which such loss of cytoplasm (and activity) proceeds can be expressed as a linear function of the specific growth rate. At growth rates above some minimum value (0.0115 h(-1)) cell degeneration can be prevented. This model served as the control basis during open-loop as well as closed-loop computer control of the fermentation. Closed-loop control was achieved through feedback information of biomass concentration using a filtration probe and was required when complex nutrients contributed significantly to the overall biomass production.