Showing papers on "Bioprocess published in 2012"

Bioreactor monitoring with spectroscopy and chemometrics: a review

Abstract: Biotechnological processes are crucial to the development of any economy striving to ensure a relevant position in future markets. The cultivation of microorganisms in bioreactors is one of the most important unit operations of biotechnological processes, and real-time monitoring of bioreactors is essential for effective bioprocess control. In this review, published material on the potential application of different spectroscopic techniques for bioreactor monitoring is critically discussed, with particular emphasis on optical fiber technology, reported for in situ bioprocess monitoring. Application examples are presented by spectroscopy type, specifically focusing on ultraviolet–visible, near-infrared, mid-infrared, Raman, and fluorescence spectroscopy. The spectra acquisition devices available and the major advantages and disadvantages of each spectroscopy are discussed. The type of information contained in the spectra and the available chemometric methods for extracting that information are also addressed, including wavelength selection, spectra pre-processing, principal component analysis, and partial least-squares. Sample handling techniques (flow and sequential injection analysis) that include transport to spectroscopic sensors for ex-situ on-line monitoring are not covered in this review.

Soft sensors in bioprocessing: A status report and recommendations

Abstract: The following report with recommendations is the result of an expert panel meeting on soft sensor applications in bioprocess engineering that was organized by the Measurement, Monitoring, Modelling and Control (M3C) Working Group of the European Federation of Biotechnology - Section of Biochemical Engineering Science (ESBES). The aim of the panel was to provide an update on the present status of the subject and to identify critical needs and issues for the furthering of the successful development of soft sensor methods in bioprocess engineering research and for industrial applications, in particular with focus on biopharmaceutical applications. It concludes with a set of recommendations, which highlight current prospects for the extended use of soft sensors and those areas requiring development.

Bioprocess development for mass production of size-controlled human pluripotent stem cell aggregates in stirred suspension bioreactor.

Abstract: Current protocols for the scalable suspension culture of human pluripotent stem cells (hPSCs) are limited by multiple biological and technical challenges that need to be addressed before their use in clinical trials. To overcome these challenges, we have developed a novel bioprocess platform for large-scale expansion of human embryonic and induced pluripotent stem cell lines as three-dimensional size-controlled aggregates. This novel bioprocess utilizes the stepwise optimization of both static and dynamic suspension culture conditions. After screening eight xeno-free media in static suspension culture and optimizing single-cell passaging in dynamic conditions, the scale-up from a static to a dynamic suspension culture in the stirred bioreactor resulted in a two- to threefold improvement in expansion rates, as measured by cell counts and metabolic activity. We successfully produced size-specific aggregates through optimization of bioreactor hydrodynamic conditions by using combinations of different agitati...

Butanol production from lignocellulosics

Abstract: Clostridium spp. produce n-butanol in the acetone/butanol/ethanol process. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, the low product yield, toxicity to production strain, multiple-end products and downstream processing of alcohol mixtures. This review describes the use of lignocellulosic feedstocks, bioprocess and metabolic engineering, downstream processing and catalytic refining of n-butanol.

Automatic control of bioprocesses.

Abstract: In this chapter, different approaches for open-loop and closed-loop control applied in bioprocess automation are discussed. Although in recent years many contributions dealing with closed-loop control have been published, only a minority were actually applied in real bioprocesses, the majority being simulations. As a result of the diversity of bioprocess requirements, a single control algorithm cannot be applied in all cases; rather, different approaches are necessary. Most publications combine different closed-loop control techniques to construct hybrid systems. These systems are supposed to combine the advantages of each approach into a well-performing control strategy. The majority of applications are soft sensors in combination with a proportional-integral-derivative (PID) controller. The fact that soft sensors have become this importance for control purposes demonstrates the lack of direct measurements or their large additional expense for robust and reliable online measurement systems. The importance of model predictive control is increasing; however, reliable and robust process models are required, as well as very powerful computers to address the computational needs. The lack of theoretical bioprocess models is compensated by hybrid systems combining theoretical models, fuzzy logic, and/or artificial neural network methodology. Although many authors suggest a possible transfer of their presented control application to other bioprocesses, the algorithms are mostly specialized to certain organisms or certain cultivation conditions as well as to a specific measurement system.

Bioprocess engineering principles.

Abstract: This welcome new edition discusses bioprocess engineering from the perspective of biology students. It includes a great deal of new material and has been extensively revised and expanded. These updates strengthen the book and maintain its position as the book of choice for senior undergraduates and graduates seeking to move from biochemistry/microbiology/molecular biology to bioprocess engineering. New to this edition: * All chapters thoroughly revised for current developments, with over 200 pgs of new material, including significant new content in: * Metabolic Engineering * Sustainable Bioprocessing * Membrane Filtration * Turbulence and Impeller Design * Downstream Processing * Oxygen Transfer Systems * Over 150 new problems and worked examples * More than 100 new illustrations

An automated workflow for enhancing microbial bioprocess optimization on a novel microbioreactor platform

Abstract: Background: High-throughput methods are widely-used for strain screening effectively resulting in binary information regarding high or low productivity. Nevertheless achieving quantitative and scalable parameters for fast bioprocess development is much more challenging, especially for heterologous protein production. Here, the nature of the foreign protein makes it impossible to predict the, e.g. best expression construct, secretion signal peptide, inductor concentration, induction time, temperature and substrate feed rate in fed-batch operation to name only a few. Therefore, a high number of systematic experiments are necessary to elucidate the best conditions for heterologous expression of each new protein of interest. Results: To increase the throughput in bioprocess development, we used a microtiter plate based cultivation system (Biolector) which was fully integrated into a liquid-handling platform enclosed in laminar airflow housing. This automated cultivation platform was used for optimization of the secretory production of a cutinase from Fusarium solani pisi with Corynebacterium glutamicum. The online monitoring of biomass, dissolved oxygen and pH in each of the microtiter plate wells enables to trigger sampling or dosing events with the pipetting robot used for a reliable selection of best performing cutinase producers. In addition to this, further automated methods like media optimization and induction profiling were developed and validated. All biological and bioprocess parameters were exclusively optimized at microtiter plate scale and showed perfect scalable results to 1 L and 20 L stirred tank bioreactor scale. Conclusions: The optimization of heterologous protein expression in microbial systems currently requires extensive testing of biological and bioprocess engineering parameters. This can be efficiently boosted by using a microtiter plate cultivation setup embedded into a liquid-handling system, providing more throughput by parallelization and automation. Due to improved statistics by replicate cultivations, automated downstream analysis, and scalable process information, this setup has superior performance compared to standard microtiter plate cultivation.

Nano-tubular cellulose for bioprocess technology development.

Abstract: Delignified cellulosic material has shown a significant promotional effect on the alcoholic fermentation as yeast immobilization support. However, its potential for further biotechnological development is unexploited. This study reports the characterization of this tubular/porous cellulosic material, which was done by SEM, porosimetry and X-ray powder diffractometry. The results showed that the structure of nano-tubular cellulose (NC) justifies its suitability for use in “cold pasteurization” processes and its promoting activity in bioprocessing (fermentation). The last was explained by a glucose pump theory. Also, it was demonstrated that crystallization of viscous invert sugar solutions during freeze drying could not be otherwise achieved unless NC was present. This effect as well as the feasibility of extremely low temperature fermentation are due to reduction of the activation energy, and have facilitated the development of technologies such as wine fermentations at home scale (in a domestic refrigerator). Moreover, NC may lead to new perspectives in research such as the development of new composites, templates for cylindrical nano-particles, etc.

One-pot bioethanol production from cellulose by co-culture of Acremonium cellulolyticus and Saccharomyces cerevisiae.

Abstract: Background While the ethanol production from biomass by consolidated bioprocess (CBP) is considered to be the most ideal process, simultaneous saccharification and fermentation (SSF) is the most appropriate strategy in practice. In this study, one-pot bioethanol production, including cellulase production, saccharification of cellulose, and ethanol production, was investigated for the conversion of biomass to biofuel by co-culture of two different microorganisms such as a hyper cellulase producer, Acremonium cellulolyticus C-1 and an ethanol producer Saccharomyces cerevisiae. Furthermore, the operational conditions of the one-pot process were evaluated for maximizing ethanol concentration from cellulose in a single reactor.

Rational bioprocess design for human pluripotent stem cell expansion and endoderm differentiation based on cellular dynamics.

Abstract: We present a predictive bioprocess design strategy employing cell- and molecular-level analysis of rate-limiting steps in human pluripotent stem cell (hPSC) expansion and differentiation, and apply it to produce definitive endoderm (DE) progenitors using a scalable directed-differentiation technology. We define a bioprocess optimization parameter (L; targeted cell Loss) and, with quantitative cell division tracking and fate monitoring, identify and overcome key suspension bioprocess bottlenecks. Adapting process operating conditions to pivotal parameters (single cell survival and growth rate) in a cell-line specific manner enabled adherent-equivalent expansion of hPSCs in feeder- and matrix-free defined-medium suspension culture. Predominantly instructive differentiation mechanisms were found to underlie a subsequent 18-fold expansion, during directed differentiation, to high-purity DE competent for further commitment along pancreatic and hepatic lineages. This study demonstrates that iPSC expansion and differentiation conditions can be prospectively specified to guide the enhanced production of target cells in a scale-free directed differentiation system.

Implementation of proton transfer reaction‐mass spectrometry (PTR‐MS) for advanced bioprocess monitoring

Abstract: We report on the implementation of proton transfer reaction-mass spectrometry (PTR-MS) technology for on-line monitoring of volatile organic compounds (VOCs) in the off-gas of bioreactors. The main part of the work was focused on the development of an interface between the bioreactor and an analyzer suitable for continuous sampling of VOCs emanating from the bioprocess. The permanently heated sampling line with an inert surface avoids condensation and interaction of volatiles during transfer to the PTR-MS. The interface is equipped with a sterile sinter filter unit directly connected to the bioreactor headspace, a condensate trap, and a series of valves allowing for dilution of the headspace gas, in-process calibration, and multiport operation. To assess the aptitude of the entire system, a case study was conducted comprising three identical cultivations with a recombinant E. coli strain, and the volatiles produced in the course of the experiments were monitored with the PTR-MS. The high reproducibility of the measurements proved that the established sampling interface allows for reproducible transfer of volatiles from the headspace to the PTR-MS analyzer. The set of volatile compounds monitored comprises metabolites of different pathways with diverse functions in cell physiology but also volatiles from the process matrix. The trends of individual compounds showed diverse patterns. The recorded signal levels covered a dynamic range of more than five orders of magnitude. It was possible to assign specific volatile compounds to distinctive events in the bioprocess. The presented results clearly show that PTR-MS was successfully implemented as a powerful bioprocess-monitoring tool and that access to volatiles emitted by the cells opens promising perspectives in terms of advanced process control. Biotechnol. Bioeng. 2012; 109: 3059–3069. © 2012 Wiley Periodicals, Inc.

Optimization of the Nutritional Parameters for Enhanced Production of B. subtilis SPB1 Biosurfactant in Submerged Culture Using Response Surface Methodology

Abstract: Nutritional requirements can contribute considerably to the production cost and the bioprocess economics. Media optimisation using response surface methodology is one of the used methods to ameliorate the bioprocess economics. In the present study, biosurfactant production by Bacillus subtilis SPB1 was effectively enhanced by response surface methodology. A Plackett-Burman-based statistical screening procedure was adopted to determine the most important factor affecting lipopeptide production. Eleven variables are screened and results show that glucose, K2HPO4, and urea concentrations influence the most biosurfactant production. A Central Composite Design was conducted to optimize the three selected factors. Statistical analyses of the data of model fitting were done by using NemrodW. Results show a maximum predicted biosurfactant concentration of 2.93 (±0.32) g/L when using 15 g/L glucose, 6 g/L urea, and 1 g/L K2HPO4. The predicted value is approximately 1.65 much higher than the original production determined by the conventional one-factor-at-a-time optimization method.

Examining the feasibility of bulk commodity production in Escherichia coli.

Abstract: Escherichia coli is currently used by many research institutions and companies around the world as a platform organism for the development of bio-based production processes for bulk biochemicals. A given bulk biochemical bioprocess must be economically competitive with current production routes. Ideally the viability of each bioprocess should be evaluated prior to commencing research, both by metabolic network analysis (to determine the maximum theoretical yield of a given biocatalyst) and by techno-economic analysis (TEA; to determine the conditions required to make the bioprocess cost-competitive). However, these steps are rarely performed. Here we examine theoretical yields and review available TEA for bulk biochemical production in E. coli. In addition, we examine fermentation feedstocks and review recent strain engineering approaches to achieve industrially-relevant production, using examples for which TEA has been performed: ethanol, poly-3-hydroxybutyrate, and 1,3-propanediol.

Recovery of Native Potato Protein Comparing Expanded Bed Adsorption and Ultrafiltration

Abstract: This is the postprint version of the article published in Food and Bioprocess Technology. The published article is available at www.springerlink.com

Multivariate Data Analysis for Advancing the Interpretation of Bioprocess Measurement and Monitoring Data

Abstract: The advances in measurement techniques, the growing use of high-throughput screening and the exploitation of ‘omics’ measurements in bioprocess development and monitoring increase the need for effective data pre-processing and interpretation. The multi-dimensional character of the data requires the application of advanced multivariate data analysis (MVDA) tools. An overview of both linear and non-linear MVDA tools most frequently used in bioprocess data analysis is presented. These include principal component analysis (PCA), partial least squares (PLS) and their variants as well as various types of artificial neural networks (ANNs). A brief description of the basic principles of each of the techniques is given with emphasis on the possible application areas within bioprocessing and relevant examples.

Microscale bioprocessing system and method for protein manufacturing

Abstract: A bioprocessing system for protein manufacturing is provided that is compact, integrated and suited for on-demand production and delivery of therapeutic proteins to patients. The system can also be used for efficient on-demand production of any type of protein.

Advances and trends in the design, analysis, and characterization of polymer-protein conjugates for "PEGylaided" bioprocesses.

Abstract: In addition to their use as therapeutics and be- cause of their enhanced properties, PEGylated proteins have potential application in fields such as bioprocessing. How- ever, the use of PEGylated conjugates to improve the per- formance of bioprocess has not been widely explored. This limited additional industrial use of PEG-protein conjugates can be attributed to the fact that PEGylation reactions, separation of the products, and final characterization of the structure and activity of the resulting species are not trivial tasks. The development of bioprocessing operations based on PEGylated proteins relies heavily in the use of analytical tools that must sometimes be adapted from the strategies used in pharmaceutical conjugate development. For in- stance, to evaluate conjugate performance in bioprocessing operations, both chromatographic and non-chromatographic steps must be used to separate and quantify the resulting reaction species. Characterization of the conjugates by mass spectrometry, circular dichroism, and specific activity assays, among other adapted techniques, is then required to evaluate the feasibility of using the conjugates in any operation. Correct selection of the technical and analytical methods in each of the steps from design of the PEGylation reaction to its final engineering application will ensure suc- cess in implementing a "PEGylaided" process. In this con- text, the objective of this review is to describe technological and analytical trends in developing successful applications of PEGylated conjugates in bioprocesses and to describe potential fields in which these proteins can be exploited.

Mini-scale bioprocessing systems for highly parallel animal cell cultures

Abstract: Animal cells have been used extensively in therapeutic protein production. The growth of animal cells and the expression of therapeutic proteins are highly dependent on the culturing environments. A large number of experimental permutations need to be explored to identify the optimal culturing conditions. Miniaturized bioreactors are well suited for such tasks as they offer high-throughput parallel operation and reduce cost of reagents. They can also be automated and be coupled to downstream analytical units for online measurements of culture products. This review summarizes the current status of miniaturized bioreactors for animal cell cultivation based on the design categories: microtiter plates, flasks, stirred tank reactors, novel designs with active mixing, and microfluidic cell culture devices. We compare cell density and product titer, for batch or fed-batch modes for each system. Monitoring/controlling devices for engineering parameters such as pH, dissolved oxygen, and dissolved carbon dioxide, which could be applied to such systems, are summarized. Finally, mini-scale tools for process performance evaluation for animal cell cultures are discussed: total cell density, cell viability, product titer and quality, substrates, and metabolites profiles.

Bioprocess Engineering: Kinetics, Biosystems, Sustainability, and Reactor Design

Abstract: "Bioprocess Engineering" involves the design and development of equipment and processes for the manufacturing of products such as food, feed, pharmaceuticals, nutraceuticals, chemicals, and polymers and paper from biological materials It also deals with studying various biotechnological processes "Bioprocess Kinetics and Systems Engineering" first of its kind contains systematic and comprehensive content on bioprocess kinetics, bioprocess systems, sustainability and reaction engineering Dr Shijie Liu reviews the relevant fundamentals of chemical kinetics-including batch and continuous reactors, biochemistry, microbiology, molecular biology, reaction engineering, and bioprocess systems engineering - introducing key principles that enable bioprocess engineers to engage in the analysis, optimization, design and consistent control over biological and chemical transformations The quantitative treatment of bioprocesses is the central theme of this book, while more advanced techniques and applications are covered with some depth Many theoretical derivations and simplifications are used to demonstrate how empirical kinetic models are applicable to complicated bioprocess systems This title contains extensive illustrative drawings which make the understanding of the subject easy It also contains worked examples of the various process parameters, their significance and their specific practical use It provides the theory of bioprocess kinetics from simple concepts to complex metabolic pathways It incorporates sustainability concepts into the various bioprocesses

Method and apparatus for growing microbial cultures that require gaseous electron donors, electron acceptors, carbon sources, or other nutrients

Abstract: Compositions and methods and apparatus for growth and maintenance of microorganisms and/or bioprocesses using one or more gases as electron donors, electron acceptors, carbon sources, or other nutrients, and for a bioprocess that converts hydrogen and carbon dioxide, or syngas, or producer gas into lipid products, bio-based oils, or other biochemical products.

Potential applications of bioprocess technology in petroleum industry

Abstract: Petroleum refining is traditionally based on the use of physicochemical processes such as distillation and chemical catalysis that operate under high temperatures and pressures conditions, which are energy intensive and costly. Biotechnology has become an important tool for providing new approaches in petroleum industry during oil production, refining and processing as well as managing environmentally safe pollutant remediation and disposal practices. Earlier biotechnology applications in the petroleum industry were limited to microbial enhanced oil recovery, applications of bioremediation to contaminated marine shorelines, soils and sludges. The potential role of bioprocess technology in this industry has now expanded further into the areas of biorefining and upgrading of fuels, production of fine chemicals, control of souring during production and air VOC biofiltration. In this paper we provide an overview of the major applications of bioprocesses and technology development in the petroleum industry both in upstream and downstream areas and highlight future challenges and opportunities.