Biotechnology and Bioengineering

Cover image for Vol. 110 Issue 8

August 2013

Volume 110, Issue 8

Pages C1–C1, fmi–fmvi, 2063–2315

  1. Cover

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
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      Biotechnology and Bioengineering: Volume 110, Number 8, August 2013 (page C1)

      Article first published online: 25 JUN 2013 | DOI: 10.1002/bit.24680

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      Cover Legend SEM image of early neurons derived from ES cells 48H after neuronal differentiation induction on laminin coated PDMS nanopillars 500 nm width and 200 nm height.(Image courtesy of Dr. Elisa Migliorini et. al.) .

  2. Contents

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
  3. Spotlights

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
  4. Reviews

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
    1. Microfluidic culture models to study the hydrodynamics of tumor progression and therapeutic response (pages 2063–2072)

      Cara Buchanan and Marissa Nichole Rylander

      Article first published online: 15 JUN 2013 | DOI: 10.1002/bit.24944

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      Microfluidic culture models represent the next generation of clinically relevant platforms for studying flow-mediated mechanisms of tumor progression and therapeutic response in vitro. Such systems have the potential to aid in the development of improved treatment modalities by supporting the investigation of tumor cell response to various hydrodynamic conditions or therapeutic strategies within well-defined, highly controllable, and physiologically relevant environments.

  5. Articles

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
    1. Escherichia coli-based cell free production of flagellin and ordered flagellin display on virus-like particles (pages 2073–2085)

      Yuan Lu, John P. Welsh, Wei Chan and James R. Swartz

      Article first published online: 1 MAY 2013 | DOI: 10.1002/bit.24903

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      Bacterial flagellin as a potential vaccine adjuvant has been successfully synthesized in a cell-free protein synthesis (CFPS) system. The product has much higher affinity for the TLR5 receptor (EC50 = 2.4 ± 1.4 pM) than previously reported. Ordered and oriented flagellin display on virus-like particles (VLPs) increased TLR5 stimulation by approximately 10-fold.

    2. Quantitating intraparticle O2 gradients in solid supported enzyme immobilizates: Experimental determination of their role in limiting the catalytic effectiveness of immobilized glucose oxidase (pages 2086–2095)

      Juan M. Bolivar, Tanja Consolati, Torsten Mayr and Bernd Nidetzky

      Article first published online: 7 MAR 2013 | DOI: 10.1002/bit.24873

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      Using a new optical sensing method for real-time measurement of the internal O2 concentration in porous enzyme carriers (Sepabeads), it is shown that the catalytic effectiveness factors for different immobilizates of glucose oxidase were strongly limited by intraparticle O2 availability. Correction for this internal diffusion limitation allowed for determination of the intrinsic activity of the immobilized enzyme. The results inform the rational design of an optimally active oxidation biocatalyst on solid support.

    3. A quantitative analysis of microalgal lipids for optimization of biodiesel and omega-3 production (pages 2096–2104)

      Ian L.D. Olmstead, David R.A. Hill, Daniel A. Dias, Nirupama S. Jayasinghe, Damien L. Callahan, Sandra E. Kentish, Peter J. Scales and Gregory J.O. Martin

      Article first published online: 9 FEB 2013 | DOI: 10.1002/bit.24844

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      The design of optimized processes for converting algal biomass to biodiesel requires detailed understanding of the lipid content of different algal species and how this varies under different growth conditions. While many techniques are available for lipid analysis, most are not able to provide the quantitative information required for this purpose. In this paper the authors outline a method for a detailed characterization of microalgal lipids that is targeted towards optimization of biodiesel production.

    4. Production of polymalic acid and malic acid by Aureobasidium pullulans fermentation and acid hydrolysis (pages 2105–2113)

      Xiang Zou, Yipin Zhou and Shang-Tian Yang

      Article first published online: 16 MAR 2013 | DOI: 10.1002/bit.24876

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      A novel fermentation process with immobilized cells of Aureobasidium pullulans ZX-10 in a fibrous bed bioreactor (FBB) was developed to produce polymalic acid (PMA) at a high malic acid titer of 144 g/L and productivity of 0.74 g/L·h. Malic acid is obtained after acid hydrolysis of PMA isolated and purified by alcohol precipitation and anion-exchange adsorption. This process provides an efficient and economical way for PMA and malic acid production from glucose.

    5. Investigation of biomass concentration, lipid production, and cellulose content in Chlorella vulgaris cultures using response surface methodology (pages 2114–2122)

      Ana-Maria Aguirre and Amarjeet Bassi

      Article first published online: 4 MAR 2013 | DOI: 10.1002/bit.24871

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      This study addresses the effect of culture conditions, specifically carbon dioxide, and sodium nitrate concentrations, on biomass concentration and the ratio of lipid productivity/cellulose content. Optimization of these variables was done applying response surface methodology and the models were experimentally validated. Differences on cellulose content in cells of Chlorella vulgaris were obtained when subjected to different culture conditions, and the location of an optimal point in the range of study, where lipid productivity is high and cellulose content is low, was possible by means of the central composite design.

    6. Enzyme production by the mixed fungal culture with nano-shear pretreated biomass and lignocellulose hydrolysis (pages 2123–2130)

      Jue Lu, Rankothge Ranjith Weerasiri, Yan Liu, Wei Wang, Shaowen Ji and Ilsoon Lee

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24883

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      This work studied enzyme production on novel nano-shear pretreated corn stover by the mixed fungi culture of Trichoderma reesei (T. reesei) RUT-C30 and Aspergillus niger (A. niger). Enzymes with higher cellulolytic and xylanolytic activities were produced on corn stover pretreated with a nano-shear mixing reactor, in comparison with other pretreatment methods. Premixing of corn stover in the culture medium before nano-shear pretreatment is contributed to leave out any downstream treatment, which makes nano-shear pretreatment an efficient and economically feasible approach.

    7. Monitoring anaerobic sequential batch reactors via fractal analysis of pH time series (pages 2131–2139)

      H.O. Méndez-Acosta, E. Hernandez-Martinez, J.A. Jáuregui-Jáuregui, J. Alvarez-Ramirez and H. Puebla

      Article first published online: 29 JAN 2013 | DOI: 10.1002/bit.24838

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      Fractal analysis can be used as an effective tool for on-line monitoring of the evolution of key process variables such as COD, total VFA and the biogas production in anaerobic sequential batch reactors (AnSBR). The fact that the evolution of such variables can be followed with a simple pH measurement is of special interest for industrial application, and opens a wide rage of possibilities for the proposal of advanced but inexpensive monitoring and control schemes.

    8. Enhanced algal growth rate in a Taylor vortex reactor (pages 2140–2149)

      Bo Kong, Jacqueline V. Shanks and R. Dennis Vigil

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24886

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      C. vulgaris growth curves for experiments carried out in a batch Taylor vortex algal photobioreactor demonstrate significant increases in biomass production rate with increasing cylinder rotation speeds. Mass transport effects alone cannot explain this increased productivity whereas these results are consistent with the flashing light effect. Symbols represent experimental measurements of biomass (dry weight) concentration for cylinder rotation rates Ω = 0 rpm (○), 500 rpm (□), 1,000 rpm (×), and 1,500 rpm (◊).

    9. An ultra scale-down approach to study the interaction of fermentation, homogenization, and centrifugation for antibody fragment recovery from rec E. coli (pages 2150–2160)

      Qiang Li, Gareth J. Mannall, Shaukat Ali and Mike Hoare

      Article first published online: 7 MAY 2013 | DOI: 10.1002/bit.24891

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      Ultra scale-down tools may be used to study the complex relationship, which exists at manufacturing scale, between fermentation, cell disruption and clarification by centrifugation. The understanding of this relationship is critical if recombinant proteins e.g., antibody fragments, are to be effectively produced and recovered in a clarified suspension suitable for subsequent high resolution purification. As fermentation time progresses the producing E. coli cells become progressively easier to disrupt but yield disruptates which are ever more difficult to clarify.

    10. Semi-continuous in situ magnetic separation for enhanced extracellular protease production—modeling and experimental validation (pages 2161–2172)

      Martin Cerff, Alexander Scholz, Tobias Käppler, Kim E. Ottow, Tim J. Hobley and Clemens Posten

      Article first published online: 31 MAR 2013 | DOI: 10.1002/bit.24893

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      Semi-continuous in situ magnetic separation (ISMS) of extracellular protease increased the overall protease yield of the bioprocess up to 98% (compared to none-ISMS processes) when six subsequent separation steps were applied. Production limitations emerge from proteolysis of the target in the bioreactor and are minimized by ISMS. Hereby, the protease was captured by bacitracin-functionalized magnetic particles from the biosuspension and eluted separately. Capture of the beads was conducted by high gradient magnetic separation.

    11. Increased curvature of hollow fiber membranes could up-regulate differential functions of renal tubular cell layers (pages 2173–2183)

      Chong Shen, Qin Meng and Guoliang Zhang

      Article first published online: 31 MAR 2013 | DOI: 10.1002/bit.24874

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      The increased curvature of polysulfone membranes with or without polyethylene glycol modification could upregulate the renal cell functions without altering the confluent morphology of renal cells adhered on the surfaces. Such effect was contributed by the increased mechanical stress in the renal cells via approaching membrane curvature to that in a kidney tubule. This study will expand the concept on mimicking the in vivo microenvironment and facilitate the membrane application in tissue engineering.

    12. Non-invasive UPR monitoring system and its applications in CHO production cultures (pages 2184–2194)

      Zhimei Du, David Treiber, Rebecca E. McCoy, Amanda K. Miller, Mei Han, Feng He, Sarah Domnitz, Carole Heath and Pranhitha Reddy

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24877

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      UPR is highly activated during production culture. Stable clones obtaining the UPR-inducible system that derived from either a CHO host or a mAb-expressing cell line were either subcultured in Amgen in-house media or subjected to a 10-day fed-batch production culture side by side. UPR-induced dGFP expression levels of both subculture and day 6 production culture were measured simultaneously by fluorescence microscopy.

    13. Anti-cell death engineering of CHO cells: Co-overexpression of Bcl-2 for apoptosis inhibition, Beclin-1 for autophagy induction (pages 2195–2207)

      Jae Seong Lee, Tae Kwang Ha, Jin Hyoung Park and Gyun Min Lee

      Article first published online: 16 MAR 2013 | DOI: 10.1002/bit.24879

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      In order to achieve a more efficient protection of cells from stressful culture conditions, the simultaneous targeting of anti-apoptosis and pro-autophagy in CHO cells was attempted by co-overexpressing an anti-apoptotic protein, Bcl-2, and a key regulator of autophagy pathway, Beclin-1, respectively. This approach achieved a more efficient inhibition of cell death and a longer culture period, providing the evidence that pro-autophagy engineering together with anti-apoptosis engineering yields a synergistic effect and successfully enhances the anti-cell death engineering of CHO cells.

    14. Cooverexpression of alanine aminotransferase 1 in Chinese hamster ovary cells overexpressing taurine transporter further stimulates metabolism and enhances product yield (pages 2208–2215)

      Hisahiro Tabuchi and Tomoya Sugiyama

      Article first published online: 16 MAR 2013 | DOI: 10.1002/bit.24881

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      The host cell engineering strategy from Tabuchi and Sugiyama using TAUT increased the productive culture period from 14 days to 31 days and increased the product concentration at harvest from 3.9 g/L to 8.1 g/L. Strains co-overexpressing TAUT and ALT1 shortened the culture period and increased the product concentration at harvest from 3.9 g/L/d14 to 5.9 g/L/d14.

    15. Improved polyhydroxybutyrate production by Saccharomyces cerevisiae through the use of the phosphoketolase pathway (pages 2216–2224)

      Kanokarn Kocharin, Verena Siewers and Jens Nielsen

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24888

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      This study demonstrates pathway engineering of central carbon metabolism aiming to increase cytosolic acetyl-CoA and NADPH supply for polyhydroxybutyrate (PHB) production in Saccharomyces cerevisiae. Overexpression of endogenous genes in the ethanol degradation pathway and expression of the heterologous phosphoketolase pathway were evaluated as alternative sources for cytosolic acetyl-CoA used for PHB production. Heterologous expression of bacterial gapN to regenerate NADPH was carried out. The assessment of combined strategies reveals that the availability of cytosolic acetyl-CoA and NADPH influence PHB production.

    16. Characterization of three loci for homologous gene targeting and transgene expression (pages 2225–2235)

      Justin Eyquem, Laurent Poirot, Roman Galetto, Andrew M. Scharenberg and Julianne Smith

      Article first published online: 2 MAY 2013 | DOI: 10.1002/bit.24892

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      To identify potential integration sites for efficient transgene expression, the authors performed homologous gene targeting at three new loci chosen to represent different genomic environments. For clones carrying single copy insertions, they observed sustainable expression of a GFP cassette without disturbing the expression of flanking genes. Differences in expression were observed as a function of the locus, the promoter and the transgene orientation within the cassette but, the expression levels observed among individual clones with the same expression cassette at the same locus were highly reproducible and predictable.

    17. Polymer peel-off mask for high-resolution surface derivatization, neuron placement and guidance (pages 2236–2241)

      Dolores Martinez, Christophe Py, Mike Denhoff, Robert Monette, Tanya Comas, Anthony Krantis and Geoffrey Mealing

      Article first published online: 29 MAY 2013 | DOI: 10.1002/bit.24887

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      The effect on process guidance for high resolution patterning is described. Combined GFAP and MAP-2 fluorescence images of brain cells at 14 DIV following immunostaining on 10 mm patterns are given. Processes are shown to follow PLL patterns.

    18. Stoichiometric modeling of oxidation of reduced inorganic sulfur compounds (Riscs) in Acidithiobacillus thiooxidans (pages 2242–2251)

      Roberto A. Bobadilla Fazzini, Maria Paz Cortés, Leandro Padilla, Daniel Maturana, Marko Budinich, Alejandro Maass and Pilar Parada

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24875

    19. Distributed modeling of human influenza a virus–host cell interactions during vaccine production (pages 2252–2266)

      Thomas Müller, Robert Dürr, Britta Isken, Josef Schulze-Horsel, Udo Reichl and Achim Kienle

      Article first published online: 7 APR 2013 | DOI: 10.1002/bit.24878

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      During influenza vaccine production in mammalian cell cultures the progress of infection can be characterized by the intracellular amount of virus proteins which is measured via flow cytometry. This allows the differentiation of the host cell population and reveals characteristic dynamic phenomena of the cell distribution like transient multimodality and reversal of propagation direction. With the presented population balance model the authors explicitly address these phenomena for the first time for different influenza virus subtypes by utilizing a distributed modeling approach to gain deeper insight into the underlying kinetic processes.

    20. Segregated flux balance analysis constrained by population structure/function data: The case of PHA production by mixed microbial cultures (pages 2267–2276)

      F. Pardelha, M.G.E. Albuquerque, G. Carvalho, M.A.M. Reis, J.M.L. Dias and R. Oliveira

      Article first published online: 3 MAY 2013 | DOI: 10.1002/bit.24894

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      A segregated FBA method was developed and applied to a mixed microbial culture (MMC) that produces PHA from fermented sugar cane molasses. This method enables computation population specific flux vectors, therewith enabling profiling of the metabolic heterogeneity within a MMC. The segregated FBA is shown to accurately predict the average PHA storage flux and the respective monomeric composition for 16 independent experiments. Moreover, the predicted fluxes are shown to be highly concordant with MFA estimated fluxes.

    21. Laser exposure of gold nanorods can increase neuronal cell outgrowth (pages 2277–2291)

      Chiara Paviolo, John W. Haycock, Jiawey Yong, Aimin Yu, Paul R. Stoddart and Sally L. McArthur

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24889

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      Picture of NG108-15 neuronal cells cultured with gold nanorods and irradiated with a 780 nm lase is shown. Laser exposure of NRs in NG108-15 neuronal cells can stimulate differentiation, particularly with regard to an increase in neurite outgrowth. This result opens up new possibilities not only for peripheral nerve regeneration treatments, but also for novel central nervous system intervention following spinal cord injury.

    22. Dual factor delivery of CXCL12 and Exendin-4 for improved survival and function of encapsulated beta cells under hypoxic conditions (pages 2292–2300)

      Stephanie Duncanson and Athanassios Sambanis

      Article first published online: 26 MAR 2013 | DOI: 10.1002/bit.24872

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      A promising treatment for diabetes is a bioartifical pancreas. However, graft failure occurs, in part due to hypoxic conditions at the transplant site. CXCL12 and GLP-1 have been shown to have anti-apoptotic effects on beta cells, and additionally GLP-1 has insulinotropic effects. In this study, the authors found that presentation of CXCL12 in combination with delivery of Exendin-4 (Ex-4), a GLP-1 analog, significantly reduced beta-cell apoptosis and increased insulin secretion under hypoxic conditions.

    23. Nanomechanics controls neuronal precursors adhesion and differentiation (pages 2301–2310)

      Elisa Migliorini, Jelena Ban, Gianluca Grenci, Laura Andolfi, Alessandro Pozzato, Massimo Tormen, Vincent Torre and Marco Lazzarino

      Article first published online: 16 MAR 2013 | DOI: 10.1002/bit.24880

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      The effect of substrate elasticity on neuronal precursors differentiation and adhesion is described. Top: picture which represents the deformations of soft (left) and rigid (right) substrates induced by the detachment of a neuronal precursors. Bottom: IF representative images of ES-derived cell culture on the two substrates with the corresponding neuronal yield; neurons were labeled with Tuj1 (green), neuronal precursors with nestin (red) and nuclei were stained with Hoechst. The neuronal yield was calculated 48 h after neuronal plating.

  6. Communications to the Editor

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communications to the Editor
    1. Scale-up and intensification of (S)-1-(2-chlorophenyl)ethanol bioproduction: Economic evaluation of whole cell-catalyzed reduction of o-Chloroacetophenone (pages 2311–2315)

      Thomas Eixelsberger, John M. Woodley, Bernd Nidetzky and Regina Kratzer

      Article first published online: 31 MAR 2013 | DOI: 10.1002/bit.24896

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      Reduction of 300 mM o-chloroacetophenone was accomplished with E. coli cells co-expressing Candida tenuis xylose reductase and Candida boidinii formate dehydrogenase. Process boundaries with respect to biological, reaction and economic considerations were investigated in a ‘window of operation’ analysis. The operational space for economic (S)-1-(2-chlorophenyl)ethanol production was depicted a function of catalyst loading, product concentration, cell activity and substrate toxicity. Process scale-up and intensification extended the catalyst loading and economic product concentration limits and thereby shifted the process into the operational space.

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