Biotechnology and Bioengineering

Cover image for Vol. 109 Issue 8

August 2012

Volume 109, Issue 8

Pages C1–C1, fmi–fmvi, 1887–2171

  1. Cover

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. You have free access to this content
      Biotechnology and Bioengineering: Volume 109, Number 8, August 2012 (page C1)

      Version of Record online: 18 JUN 2012 | DOI: 10.1002/bit.24311

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      Cover Picture: The cover depicts an example of Silicone Porous Templated Scaffolds: capillary network inside silicone STAR™ pore structure (27μm pores, 11μm throats), with red blood cells surrounded by vascular endothelial cells (EC), and macrophages (Mø) lining walls of pores (rabbit sclera, H&E stain, 3 months, 40x objective). (Image provided by Dr. Andrew J. Marshall, Healionics).

  2. Contents

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
  3. Spotlights

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. Fed-batch microbioreactors (page fmvi)

      Version of Record online: 18 JUN 2012 | DOI: 10.1002/bit.24309

  4. Editorial

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. In memoriam—Elmer L. Gaden, Jr. (pages 1887–1888)

      Donald J. Kirwan, John L. Gainer and Giorgio Carta

      Version of Record online: 21 MAY 2012 | DOI: 10.1002/bit.24556

  5. Editors' Choice

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. Non-enzymatic palladium recovery on microbial and synthetic surfaces (pages 1889–1897)

      Amelia-Elena Rotaru, Wei Jiang, Kai Finster, Troels Skrydstrup and Rikke Louise Meyer

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24500

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      The use of microorganisms as support for reduction of dissolved Pd(II) to immobilized Pd(0)-nanoparticles is an environmentally friendly approach for Pd recovery from waste. This study proposes a non-enzymatic mechanism for bio-supported Pd(II)-reduction on microbial surfaces, since microorganisms stimulated Pd(II)-reduction independent of being alive or dead. A comparable effect was observed for amine-functionalized microparticles, which could be hampered if amine-groups were blocked by acetylation. Infrared-spectroscopy corroborate the role of amine-groups in Pd(II) sorption and reduction on microbial and synthetic surfaces.

  6. Review

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. Engineering biomaterials to integrate and heal: The biocompatibility paradigm shifts (pages 1898–1911)

      James D. Bryers, Cecilia M. Giachelli and Buddy D. Ratner

      Version of Record online: 24 MAY 2012 | DOI: 10.1002/bit.24559

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      Porous templated scaffolds (PTSs) are polymer constructs where each pore is exactly the same size, and pore interconnects are also uniform in size; with both parameters being adjustable. Consistently, the 30–40 µm dia. pore size PTS shows excellent healing, regardless of polymer composition or implant site. Hypothetically, the large numbers of macrophage observed in the 35-µm PTS are being directed toward the M2 (regenerative) phenotype. Growing circumstantial evidence suggests the controversial possibility that macrophage transdifferentiate into implant site-specific tissue cells.

  7. Articles

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. Biocatalysis, Protein Engineering, and Nanobiotechnology

      Laying out ground rules for protein-aided nanofabrication: ZnO synthesis at 70°C as a case study (pages 1912–1918)

      Sathana Kitayaporn, Weibin Zhou, Daniel T. Schwartz and François Baneyx

      Version of Record online: 24 FEB 2012 | DOI: 10.1002/bit.24466

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      The authors show that although biological buffer and protein scaffold can exert a significant effect on ZnO nucleation and growth, thermodynamic predictions and adsorption isotherms are powerful tools to understand protein-dependent morphogenetic effects under well-controlled synthesis conditions.

    2. Biofuels and Environmental Biotechnology

      Improved phosphate removal by selective sludge discharge in aerobic granular sludge reactors (pages 1919–1928)

      J.P. Bassin, M.-K.H. Winkler, R. Kleerebezem, M. Dezotti and M.C.M. van Loosdrecht

      Version of Record online: 22 FEB 2012 | DOI: 10.1002/bit.24457

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      Segregation of biomass in aerobic granular sludge SBRs offered a possibility to control PAO-GAO competition. Selective removal of biomass mainly from the GAO-rich part of the sludge bed is especially relevant at high temperatures (such as 30°C). The removal of sludge from the PAO-rich part of the sludge bed in minor proportions did not negatively affect P-removal. Higher poly-P content and higher temperatures would possibly lead to higher chemical precipitation within the granules.

    3. Consolidated bioprocessing (CBP) of AFEX™-pretreated corn stover for ethanol production using Clostridium phytofermentans at a high solids loading (pages 1929–1936)

      Mingjie Jin, Christa Gunawan, Venkatesh Balan and Bruce E. Dale

      Version of Record online: 22 FEB 2012 | DOI: 10.1002/bit.24458

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      Performances of CBP using Clostridium phytofermentans on AFEXTM pretreated corn stover at 4% glucan loading with (w/) and without (w/o) nutrients supplementation were found to be similar. This indicates that AFEXTM pretreated corn stover contains enough nutrients for C. phytofermentans to perform CBP. The major factor limiting CBP performance at high solids loading was the acetate production at a relatively high level.

    4. Radiotolerance of phosphatases of a Serratia sp.: Potential for the use of this organism in the biomineralization of wastes containing radionuclides (pages 1937–1946)

      M. Paterson-Beedle, B.C. Jeong, C.H. Lee, K.Y. Jee, W.H. Kim, J.C. Renshaw and L.E. Macaskie

      Version of Record online: 15 MAR 2012 | DOI: 10.1002/bit.24467

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      Metal phosphate deposition by Serratia sp. shows promise as a mechanism for removing radionuclides from contaminated waste waters. Howevers the efficacy of this technique depends on the stability of the organism and the phosphatase enzymes towards high doses of ionizing radiation under various chemical conditions. Radiostability tests showed that whole-cell phosphatase retained 80% of the initial activity, even after loss of cell culturability, suggesting it is feasible to use whole cells of Serratia for radionuclide removal via phosphate precipitation.

    5. Differential effects of nitrogen and sulfur deprivation on growth and biodiesel feedstock production of Chlamydomonas reinhardtii (pages 1947–1957)

      Turgay Cakmak, Pinar Angun, Yunus Emre Demiray, Alper Devrim Ozkan, Zeynep Elibol and Turgay Tekinay

      Version of Record online: 2 MAR 2012 | DOI: 10.1002/bit.24474

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      The present work reports the effects of nitrogen and sulphur deprivation on the model microalga Chlamydomonas reinhardtii, using triacylglycerol levels as a measure of biofuel feedstock production capacity. Sulfur starvation was found to be a better way of increasing biodiesel feedstock production efficiency than nitrogen starvation. Furthermore, mating type had a significant impact on the responses of C. reinhardtii to nutrient starvation.

    6. Physiological evaluation of a new Chlorella sorokiniana isolate for its biomass production and lipid accumulation in photoautotrophic and heterotrophic cultures (pages 1958–1964)

      Min-Xi Wan, Run-Min Wang, Jin-Lan Xia, Julian N. Rosenberg, Zhen-Yuan Nie, Naoko Kobayashi, George A. Oyler and Michael J. Betenbaugh

      Version of Record online: 2 MAR 2012 | DOI: 10.1002/bit.24477

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      A novel green unicellular microalgal isolate CCTCC M209220 can grow heterotrophically with glucose as the carbon source and accumulates lipid content over 50% (ww−1) dry weight, and arrived at the high biomass of 2,500mg/L while simultaneously raising the lipid productivity to 1,300mg/L. The relative neutral lipid content as a fraction of the total lipid is also much higher in heterotrophic culture as compared to photoautotrophic culture.

    7. Kinetics of adsorption, desorption, and re-adsorption of a commercial endoglucanase in lignocellulosic suspensions (pages 1965–1975)

      Q.Q. Wang, J.Y. Zhu, C.G. Hunt and H.Y. Zhan

      Version of Record online: 15 MAR 2012 | DOI: 10.1002/bit.24483

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      This study conducted quantitative kinetic modeling and in-situ and temporally resolved measurements of adsorption, desorption, and re-adsorption of a commercial endoglucanase in lignocellulosic suspensions. The study defined a cellulase adsorption and desorption competition parameter, a pseudo rate of binding and desorption, as well as cellulase binding reversibility and recyclability. Increasing temperature and pH increase cellulase desorption and increased binding reversibility and capacity.

    8. Bioprocess Engineering and Supporting Technologies

      Fed-batch microbioreactor platform for scale down and analysis of a plasmid DNA production process (pages 1976–1986)

      Diana M. Bower, Kevin S. Lee, Rajeev J. Ram and Kristala L.J. Prather

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24498

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      The authors have developed a microbioreactor with a 1-mL working volume that is capable of fed-batch operation; control of dissolved oxygen, pH, and temperature; as well as continuous monitoring of cell growth. In this work, The authors demonstrated that the micro-scale device can accurately mimic a complex, bench-scale plasmid DNA production process. They also identified key process parameters required for consistency of results across scales.

    9. Determination and control of low-level amino acid misincorporation in human thioredoxin protein produced in a recombinant Escherichia coli production system (pages 1987–1995)

      Robert P. Harris, Jo Mattocks, Philip S. Green, Frank Moffatt and Peter M. Kilby

      Version of Record online: 22 FEB 2012 | DOI: 10.1002/bit.24462

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      Here, the authors discuss the production of the potential biopharmaceutical human thioredoxin protein (rhTRX) within E. coli. In production batches they identified and quantified a subset of rhTRX proteins differing slightly in amino acid composition by using modern proteomic technology. These misincorporations's present significant problems with molecules that are destined for clinical applications. In this study, the authors show how these can be reduced and eliminated by manipulating gene sequence and scoping fermentation composition.

    10. High gas pressure: An innovative method for the inactivation of dried bacterial spores (pages 1996–2004)

      A. Colas de la Noue, V. Espinasse, J.-M. Perrier-Cornet and P. Gervais

      Version of Record online: 24 FEB 2012 | DOI: 10.1002/bit.24465

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      Dehydrated Bacillus subtilis spores (aw 0.1) were treated under nitrogen atmosphere at 430 MPa and the pressure was then quickly released until atmospheric pressure (>75 MPa s−1). The images show the progressive loss of spore phase-bright properties after rehydration, incubated for 6 days at 37°C. This slow phenomenon could be attributed to some rehydration of the core, probably due to a weak alteration of the inner membrane during the fast gas expansion. (A: control; B: treated spores).

    11. Effects of phosphorous, nitrogen, and carbon limitation on biomass composition in batch and continuous flow cultures of the heterotrophic dinoflagellate Crypthecodinium cohnii (pages 2005–2016)

      Daniel Pleissner and Niels T. Eriksen

      Version of Record online: 2 MAR 2012 | DOI: 10.1002/bit.24470

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      This paper describes biomass composition of phosphorous, nitrogen, and carbon limited continuous flow cultures of the heterotrophic dinoflagellate Crypthecodinium cohnii. Specific contents of phosphorous, protein, and starch were strongly affected by nutrient limitation, while the specific lipid content, observable by yellow fluorescence after Nile red staining, remained almost unaffected. Carbon was taken up and stored in lipids and starch whenever available. Production of lipids that are rich in docosahexaenoic acid, therefore does not depend on nutrient limitation in C. cohnii.

    12. Leukocyte counting from a small amount of whole blood using a size-controlled microcavity array (pages 2017–2024)

      Masahito Hosokawa, Marie Asami, Seita Nakamura, Tomoko Yoshino, Noriyuki Tsujimura, Masayuki Takahashi, Satoshi Nakasono, Tsuyoshi Tanaka and Tadashi Matsunaga

      Version of Record online: 24 FEB 2012 | DOI: 10.1002/bit.24471

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      A leukocyte-counting device integrated with a size-controlled microcavity array was designed for size and deformability-based separation of leukocytes from whole blood. This device enabled efficient separation of leukocytes from small amounts of blood such as finger-prick blood, and highly accurate enumeration of total numbers of leukocytes and their subsets. The accuracy and simplicity of this absolute leukocyte counting system offers a range of potential applications, including point-of-care diagnostic systems such as human CD4 T lymphocyte counting and hematotoxicity monitoring in small animals.

    13. A model for cellulase production from Trichoderma reesei in an airlift reactor (pages 2025–2038)

      Rachid Bannari, Abdelfettah Bannari, Patrick Vermette and Pierre Proulx

      Version of Record online: 5 MAR 2012 | DOI: 10.1002/bit.24473

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      This mathematical model uses state-of-the-art Computational Fluid Dynamics coupled with models for the bubbles transport, break-up and coalescence to predict the mass transfer between the broth and the air bubbles during the fermentation of Trichoderma reesei in an airlift bioreactor.

    14. Bioseparations and Downstream Processing

      Prediction and verification of centrifugal dewatering of P. pastoris fermentation cultures using an ultra scale-down approach (pages 2039–2047)

      A.G. Lopes and E. Keshavarz-Moore

      Version of Record online: 22 MAR 2012 | DOI: 10.1002/bit.24478

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      A simple and easy to use ultra scale-down (USD) method that used small quantities of feedstock (<50 ml) and readily available centrifuge tubes in a laboratory centrifuge was developed to predict dewatering performance of pilot-scale centrifuges. It enabled a rapid and accurate (P-value ≥0.05) determination of centrifugation conditions that maximized dewatering performance during processing of yeast suspensions and cultures in two industrially relevant centrifuges: a tubular bowl Carrpowerfuge™ and a disc-stack CSA-1.

    15. Monoclonal antibody capture and viral clearance by cation exchange chromatography (pages 2048–2058)

      G.R. Miesegaes, S. Lute, D.M. Strauss, E.K. Read, A. Venkiteshwaran, A. Kreuzman, R. Shah, P. Shamlou, D. Chen and K. Brorson

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24480

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      This article evaluates the use of cation exchange chromatography as an alternative capture method for monoclonal antibodies. Using a model IgG4 cell culture harvest, the authors define a reasonable but limited range of conditions that provide acceptable values for yield, purity, and viral clearance. As shown in the accompanying figure, post-elution antibody integrity was assessed, revealing that appropriate levels of percent dimer (A) monomer (B) and half-antibody (C) forms were achieved.

    16. Use of focused acoustics for cell disruption to provide ultra scale-down insights of microbial homogenization and its bioprocess impact—recovery of antibody fragments from rec E. coli (pages 2059–2069)

      Qiang Li, Jean P. Aucamp, Alison Tang, Alex Chatel and Mike Hoare

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24484

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      Focused acoustics allows the application of high rates of intensive energy input to millilitre volumes of suspension. Parallels can be drawn with industrial-scale multilitre methods for cell disruption by high pressure homogenization. The control of focused acoustics power and time of exposure is used to create an ultra scale-down method for use in bioprocess discovery. Successfully predicted are: product release from rec E. coli; particle size distribution and rheology of the disrupted cell suspension; clarification using ultra scale-down centrifugation.

    17. Cellular and Metabolic Engineering

      Improved L-lysine production with Corynebacterium glutamicum and systemic insight into citrate synthase flux and activity (pages 2070–2081)

      Jan van Ooyen, Stephan Noack, Michael Bott, Alexander Reth and Lothar Eggeling

      Version of Record online: 22 MAR 2012 | DOI: 10.1002/bit.24486

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      Multi-start optimization for flux estimation is described. The authors delineate a comparison of the initial flux samples that are even randomly distributed in the feasible flux space with the local optima solutions after convergence of the optimizer.

    18. Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae (pages 2082–2092)

      Jie Sun, Zengyi Shao, Hua Zhao, Nikhil Nair, Fei Wen, Jian-He Xu and Huimin Zhao

      Version of Record online: 15 MAR 2012 | DOI: 10.1002/bit.24481

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      A total of 14 constitutive promoters from Saccharomyces cerevisiae were cloned and characterized under varying glucose and oxygen concentrations. As an example of application for these promoters in metabolic engineering, a novel 10-gene pathway was constructed using promoters with medium and high strength, which resulted in a Saccharomyces cerevisiae strain capable of direct conversion of birchwood xylan to zeaxanthin.

    19. Determination of Chinese hamster ovary cell line stability and recombinant antibody expression during long-term culture (pages 2093–2103)

      Laura A Bailey, Diane Hatton, Ray Field and Alan J Dickson

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24485

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      Long-term culture (LTC) of rCHO cells is associated with altered cellular stress perception that has consequences for recombinant protein yield. Expression of Growth Arrest and DNA Damage gene 153 (GADD153) exhibits earlier activation during batch culture indicating altered cellular perception or response to a constant culture environment. These data reflect part of a cellular phenotypic alteration that occurs during LTC, including metabolic changes and the capacity to support recombinant protein expression.

    20. Engineering Science of Biological Systems

      The roles of RGD and grooved topography in the adhesion, morphology, and differentiation of C2C12 skeletal myoblasts (pages 2104–2115)

      Peng-Yuan Wang, Helmut Thissen and Wei-Bor Tsai

      Version of Record online: 22 FEB 2012 | DOI: 10.1002/bit.24452

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      RGD-contained peptides were chemically conjugated to nanogrooved polystyrene (PS) substrates. The spreading and alignment of C2C12 cells on PS surfaces were enhanced by RGD conjugation. The differentiation of C2C12 cells into multi-nuclei myotubes was also enhanced on RGD-conjugated grooved PS.

    21. CFD simulation of mixing for high-solids anaerobic digestion (pages 2116–2126)

      Binxin Wu

      Version of Record online: 16 MAR 2012 | DOI: 10.1002/bit.24482

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      Computational fluid dynamics simulations of mechanical mixing for high-solids anaerobic digestion are carried out for six designs. In terms of mixing time with respect to mixing power, mixing with two modified high solidity 3/39° impellers requires the lowest power input to homogenize the manure slurry. A test of mixing with one helical ribbon impeller indicates that the mixing energy varies with manure type and total solids concentration to obtain a given mixing time.

    22. Systems Biotechnology

      A computational framework for the design of optimal protein synthesis (pages 2127–2133)

      Julien Racle, Jan Overney and Vassily Hatzimanikatis

      Version of Record online: 24 FEB 2012 | DOI: 10.1002/bit.24463

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      The authors propose a computational framework to optimize the design of synthetic gene sequences for high protein synthesis rate in heterologous gene expression and for applications in synthetic biology. The framework is based on a detailed mechanistic model of translation and it takes into account genome sequence, and physicochemical and physiological constraints. The flexible formulation allows for integration of new information, various types of constraints, and scoring methods.

    23. Tissue Engineering and Delivery Systems

      Modification of macroporous titanium tracheal implants with biodegradable structures: Tracking in vivo integration for determination of optimal in situ epithelialization conditions (pages 2134–2146)

      Nihal Engin Vrana, Agnes Dupret-Bories, Charlotte Bach, Christophe Chaubaroux, Christelle Coraux, Dominique Vautier, Fouzia Boulmedais, Youssef Haikel, Christian Debry, Marie-Helene Metz-Boutigue and Philippe Lavalle

      Version of Record online: 2 MAR 2012 | DOI: 10.1002/bit.24456

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      Hierarchically porous PLLA/titanium scaffolds which can direct cell movement in vitro were shown to control cell movement in vivo and also in a tracheal replacement model. Methods based on protein characterization were utilized to determine the optimal time point for in situ epithelialization together with development of a basement membrane-like collagen/alginate multilayer structure to improve epithelialization. The monitoring of implant integration with protein characterization and improvement of epithelialization of porous titanium implants would improve their patency in tracheal replacement.

    24. Thermo-responsive non-woven scaffolds for “smart” 3D cell culture (pages 2147–2158)

      Claire L. Rossouw, Avashnee Chetty, Francis Sean Moolman, Lyn-Marie Birkholtz, Heinrich Hoppe and Dalu T. Mancama

      Version of Record online: 24 FEB 2012 | DOI: 10.1002/bit.24469

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      The authors describe the development and testing of a new “smart” scaffold for use in 3D cell culture. Two poly(propylene) non-woven scaffolds grafted with poly(N-isopropylacrylamide) were found to be both thermoresponsive, that is, able to release viable cellular aggregates from the grafted scaffolds without mechanical scraping, scaffold degradation or the use of enzymes. The scaffolds were also capable of maintaining hepatocytes with enhanced metabolic characteristics as compared to their 2D counterparts.

  8. Communications to the Editor

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editorial
    6. Editors' Choice
    7. Review
    8. Articles
    9. Communications to the Editor
    1. Biocatalysis, Protein Engineering, and Nanobiotechnology

      Experimental determination of thermodynamic equilibrium in biocatalytic transamination (pages 2159–2162)

      Pär Tufvesson, Jacob S. Jensen, Wolfgang Kroutil and John M. Woodley

      Version of Record online: 2 MAR 2012 | DOI: 10.1002/bit.24472

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      The thermodynamic equilibrium of biocatalytic transamination is a key decision point for process design. The equilibrium position varies within several orders of magnitude for different products and amine donors but no reliable quantitative values are available in the literature. This article suggests a simple experimental approach for determining the equilibrium and exemplifies with a number of industrially relevant compounds.

    2. Bioprocess Engineering and Supporting Technologies

      A new electrochemical assay method for gene expression using hela cells with a secreted alkaline phosphatase (SEAP) reporter system (pages 2163–2167)

      Mustafa Şen, Kosuke Ino, Hitoshi Shiku and Tomokazu Matsue

      Version of Record online: 22 FEB 2012 | DOI: 10.1002/bit.24461

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      A new electrochemical assay for the detection of secreted alkaline phosphatase (SEAP) from transfected HeLa cells is proposed using a microarray device and scanning electrochemical microscopy (SECM). The transfected cells were cultured in a microarray culture device covered with a substrate modified with anti-SEAP and the immobilized SEAP was then measured by SECM. The authors successfully detected the expression of SEAP from the single cells using the assay.

    3. Engineering Science of Biological Systems

      Variable electric fields for high throughput electroporation protocol design in curvilinear coordinates (pages 2168–2171)

      Francois Fernand, Liel Rubinsky, Alex Golberg and Boris Rubinsky

      Version of Record online: 15 MAR 2012 | DOI: 10.1002/bit.24479

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      The mathematical solution to the electric field equation in cylindrical coordinates, has suggested a new experimental methodology and device for reducing experimental effort in designing electroporation protocols. Using a new cylindrical electroporation system, the authors show, with an Escherichia coli cell model, how key electroporation parameters emerge precisely from single experiments rather than through interpolation from numerous experiments in the conventional Cartesian electroporation system.

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