Biotechnology and Bioengineering

Cover image for Vol. 109 Issue 9

September 2012

Volume 109, Issue 9

Pages C1–C1, fmi–fmvi, 2173–2415

  1. Cover

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

      Version of Record online: 25 JUL 2012 | DOI: 10.1002/bit.24317

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      Cover Picture: Hyperspectral confocal fluorescence imaging and multivariate curve resolution analysis of free fatty acid producing cyanobacteria reveal changes in photosynthetic pigment content (indicated by color) and subcellular pigment location. (Image courtesy of Howland D.T. Jones and Anne M. Ruffing.)

  2. Contents

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editors' Choice
    6. Articles
    7. Communication to the Editor
  3. Spotlights

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editors' Choice
    6. Articles
    7. Communication to the Editor
  4. Editors' Choice

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editors' Choice
    6. Articles
    7. Communication to the Editor
    1. In vitro 3D human small intestinal villous model for drug permeability determination (pages 2173–2178)

      Jiajie Yu, Songming Peng, Dan Luo and John C. March

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24518

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      A 3-D human intestinal model improves physiological relevance and drug absorption performance.

  5. Articles

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

      Tools for characterizing the whole-cell bio-oxidation of alkanes at microscale (pages 2179–2189)

      Chris Grant, Ana Catarina da Silva Damas Pinto, Hai-Po Lui, John M. Woodley and Frank Baganz

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24512

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      This article describes the first reported microwell whole-cell bioconversion using a water immiscible substrate that matches the specific activity and yield achieved in a 1.2 L stirred tank bioreactor. The results indicate that mass transfer rates limit productivity in the n-dodecane bio-oxidation system, rather than inherent enzyme activity.

    2. Biofuels and Environmental Biotechnology

      Physiological effects of free fatty acid production in genetically engineered Synechococcus elongatus PCC 7942 (pages 2190–2199)

      Anne M. Ruffing and Howland D.T. Jones

      Version of Record online: 9 APR 2012 | DOI: 10.1002/bit.24509

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      Cyanobacteria are attractive hosts for biofuel production; however, the production of fuel precursors like free fatty acids will affect the metabolic carbon balance within the cell. This work explores how free fatty acid production in a model cyanobacterium affects cell growth, photosynthesis, and photosynthetic pigments. These physiological effects ultimately limit the capacity for biofuel synthesis and must be addressed to enable large-scale production.

    3. Managing methanogens and homoacetogens to promote reductive dechlorination of trichloroethene with direct delivery of H2 in a membrane biofilm reactor (pages 2200–2210)

      Michal Ziv-El, Sudeep C. Popat, Katherine Cai, Rolf U. Halden, Rosa Krajmalnik-Brown and Bruce E. Rittmann

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24487

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      This study sheds light into how H2-fed biofilms, when operated to manage methanogenic and homoacetogenic activity, can be used for ex-situ bioremediation of chlorinated ethenes. Successful operation relied on controlling the pH-increase effects of methanogenesis and homoacetogenesis, and creating a H2 limitation during start-up to allow dechlorinators to compete against other microorgansims. Methanogens were additionally minimized during continuous flow operation by a limitation in bicarbonate resulting from strong homoacetogenic activity.

    4. Mineralization of pentachlorophenol with enhanced degradation and power generation from air cathode microbial fuel cells (pages 2211–2221)

      Liping Huang, Linlin Gan, Ning Wang, Xie Quan, Bruce E. Logan and Guohua Chen

      Version of Record online: 22 MAR 2012 | DOI: 10.1002/bit.24489

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      Mineralization of pentachlorophenol with enhanced degradation and power production was achieved in air cathode microbial fuel cells. Multiple parameters including the type of co-metabolism substrate, the number of diffusion layers, initial COD, initial PCP concentration, pH, and temperature affected PCP degradation, power production, and coulomic efficiency. Dominant exoelectrogens of Pseudomonas (acetate) and Klebsiella (glucose) were identified in the biofilms.

    5. Effect of light intensity on algal biomass accumulation and biodiesel production for mixotrophic strains Chlorella kessleri and Chlorella protothecoide cultivated in highly concentrated municipal wastewater (pages 2222–2229)

      Yecong Li, Wenguang Zhou, Bing Hu, Min Min, Paul Chen and Roger R. Ruan

      Version of Record online: 22 MAR 2012 | DOI: 10.1002/bit.24491

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      Algae cultivation in wastewater streams as feedstock for biofuel production can not only greatly decrease the cost involved in nutrients and water consumption but also serve the dual role of waste reduction and energy production. This paper targeted at optimized light supply to the cultivation system using concentrated municipal wastewater, and the results showed that light intensity had profound effect on biomass accumulation, wastewater nutrients removal through algae cultivation, and biodiesel productivity with mixotrophic strains from the family of Chlorella.

    6. Using electron balances and molecular techniques to assess trichoroethene-induced shifts to a dechlorinating microbial community (pages 2230–2239)

      Michal Ziv-El, Sudeep C. Popat, Prathap Parameswaran, Dae-Wook Kang, Alexandra Polasko, Rolf U. Halden, Bruce E. Rittmann and Rosa Krajmalnik-Brown

      Version of Record online: 24 APR 2012 | DOI: 10.1002/bit.24504

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      Fill-and-draw reactors inoculated with a TCE-dechlorinating consortium demonstrated a direct correlation between microbial community function and structure as the TCE-pulsing rate was increased. An electron-balance analysis predicted the community structure based on measured concentrations of products and constant net yields for each microorganism. The predictions corresponded to trends in the community structure based on pyrosequencing and qPCR up to the highest TCE pulsing rate, where deviations to the trend resulted from stress by the chlorinated ethenes.

    7. Bioprocess Engineering and Supporting Technologies

      A lectin-based gold nanoparticle assay for probing glycosylation of glycoproteins (pages 2240–2249)

      Germarie Sánchez-Pomales, Todd A. Morris, James B. Falabella, Michael J. Tarlov and Rebecca A. Zangmeister

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24513

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      The authors demonstrate a rapid glycoanalysis method by lectin binding to glycoprotein-modified gold nanoparticles detected using either simple instrumentation (e.g., DLS or spectrophotometry) or by visual inspection. The assay is applied to a model protein, RNase B and a therapeutic monoclonal antibody, rituximab. The versatility of this method is great due to the breadth of the assay design space and may prove useful for rapid, high-throughput detection of changes in glycan profiles of therapeutic glycoproteins in process development or process monitoring.

    8. Transient recombinant protein expression in a human amniocyte cell line: The CAP-T® cell system (pages 2250–2261)

      Simon Fischer, Nadine Charara, Andrea Gerber, Jens Wölfel, Gudrun Schiedner, Bernd Voedisch and Sabine Geisse

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24514

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      The excellent transfectability of CAP-T cells by lipofection is demonstrated by testing of twelve different reagents on a small scale. Apart from the individual performance of the reagents there is a striking dependence on the culture medium as well as on the transfection conditions applied.

    9. Self-cycling operation increases productivity of recombinant protein in Escherichia coli (pages 2262–2270)

      Zachary J. Storms, Tobin Brown, Dominic Sauvageau and David G. Cooper

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24492

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      A two-stage process incorporating self-cycling fermentation, a semi-continuous process inducing cell synchrony, was developed for recombinant β-galactosidase production in a temperature-sensitive phage λ/Escherichia coli model system. Using this production scheme, productivity increased 50% and production time decreased by 40% compared to a batch culture under similar conditions. In addition, the cell life cycle influenced protein synthesis: maximum protein productivity was obtained when induction was triggered immediately before cell division.

    10. Role of non-specific DNA in reducing coding DNA requirement for transient gene expression with CHO and HEK-293E cells (pages 2271–2278)

      Yashas Rajendra, Divor Kiseljak, Sagar Manoli, Lucia Baldi, David L. Hacker and Florian M. Wurm

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24494

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      The amount of plasmid DNA required for the transient production of recombinant proteins in a mammalian cell is high. Here the authors demonstrate that some of the plasmid DNA can be replaced with non-specific DNA without a corresponding loss in protein production in two hosts, CHO and HEK-293E cells. This should help to decrease the cost of large-scale transient protein production in mammalian cells.

    11. High-throughput analysis of the plasmid bioproduction process in Escherichia coli by FTIR spectroscopy (pages 2279–2285)

      Teresa Scholz, Vitor V. Lopes and Cecília R.C. Calado

      Version of Record online: 11 APR 2012 | DOI: 10.1002/bit.24502

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      This paper proposes a methodology based on FTIR spectroscopy and chemometrics for the high-throughput analysis of the plasmid bioproduction process in E. coli. Five batch cultures with different initial medium compositions are designed to represent different biomass and plasmid production behavior. The FTIR spectra are recorded from the collected cellular biomass. After spectral pre-processing, the predictive models are derived using PLS regression with the wavenumber selection performed by a Monte-Carlo strategy. The optimized models show a high coefficient of determination.

    12. Utilization of tyrosine- and histidine-containing dipeptides to enhance productivity and culture viability (pages 2286–2294)

      Sohye Kang, Johanna Mullen, Les P. Miranda and Rohini Deshpande

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24507

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      Tyrosine-containing dipeptides, such as Tyr-Lys and Tyr-His, have the capacity to enhance productivity and culture viability when added as a nutrient supplement during recombinant monoclonal antibody production in mammalian cells. Moreover, these dipeptides can facilitate improved metabolic profiles, including lower lactate and NHmath image production and better pH maintenance during fed-batch processes.

    13. Optical sensor enabled rocking T-flasks as novel upstream bioprocessing tools (pages 2295–2305)

      Jose R. Vallejos, Martina Micheletti, Kurt A. Brorson, Antonio R. Moreira and Govind Rao

      Version of Record online: 11 APR 2012 | DOI: 10.1002/bit.24508

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      This work provides the know-how principles to enable rocking T-Flasks as scale-down models of disposable wave bioreactors. It combines fluorescence-based optical sensors and engineering principles to show proof-of-concept to develop a complete disposable rocking-based process development and production platforms.

    14. Probing of C-terminal lysine variation in a recombinant monoclonal antibody production using Chinese hamster ovary cells with chemically defined media (pages 2306–2315)

      Jun Luo, Jian Zhang, Diya Ren, Wen-Lin Tsai, Feng Li, Ashraf Amanullah and Terry Hudson

      Version of Record online: 11 APR 2012 | DOI: 10.1002/bit.24510

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      The C-terminal lysine processing during monoclonal antibody production may happen both intracellularly and extracellularly with the existence and involvement of multiple sources of carboxypeptidases and could be manipulated by trace elements (i.e. copper, zinc) and their ratio.

    15. Bioseparations and Downstream Processing

      Diffusion-based extraction of DMSO from a cell suspension in a three stream, vertical microchannel (pages 2316–2324)

      Jacob Hanna, Allison Hubel and Erin Lemke

      Version of Record online: 30 MAR 2012 | DOI: 10.1002/bit.24499

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      Development of methods to remove dimethylsulfoxide (DMSO) from frozen cells will do much for improving the safety of cell therapy products and reduce adverse events. The paper describes the performance of a microfluidic device capable of removing DMSO and minimizing cell losses. In spite of being a microfluidic device, the design described in the paper is capable of processing clinical scale cell numbers in a reasonable period of time.

    16. Cellular and Metabolic Engineering

      Continuous modeling of metabolic networks with gene regulation in yeast and in vivo determination of rate parameters (pages 2325–2339)

      P. Moisset, D. Vaisman, A. Cintolesi, J. Urrutia, I. Rapaport, B.A. Andrews and J.A. Asenjo

      Version of Record online: 24 APR 2012 | DOI: 10.1002/bit.24503

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      A continuous model of a metabolic network and gene regulation in yeast using glucose and ethanol was compared with experimental data. The model is constructed using enzyme kinetics, the mechanisms of gene expression and enzyme synthesis, and data from microarrays. Data from metabolic flux analysis (MFA) is used to fit the equation parameters. The proposed method allows the obtention of reasonable parameters and concentrations in a system with a much larger number of unknowns (120) than equations (72).

    17. Cloning, mutagenesis, and characterization of the microalga Parietochloris incisa acetohydroxyacid synthase, and its possible use as an endogenous selection marker (pages 2340–2348)

      Omer Grundman, Inna Khozin-Goldberg, Dina Raveh, Zvi Cohen, Maria Vyazmensky, Sammy Boussiba and Michal Shapira

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24515

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      The cDNA that encodes acetohydroxyacid synthase of the green alga Parietochloris incisa (PiAHAS) was cloned and a W605S point mutation was introduced (PimAHAS). The recombinant mutant protein exhibited 7 orders of magnitude higher resistance to the SMM herbicide than that of the wild type. The mutant gene form can be used as an endogenous, non-antibiotic, environmentally safe selection marker for future P. incisa genetic transformation.

    18. Improvement of surfactin production in Bacillus subtilis using synthetic wastewater by overexpression of specific extracellular signaling peptides, comX and phrC (pages 2349–2356)

      Ju Jung, Kyung Ok Yu, Ahmad Bazli Ramzi, Se Hoon Choe, Seung Wook Kim and Sung Ok Han

      Version of Record online: 24 APR 2012 | DOI: 10.1002/bit.24524

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      The aim of this study is to achieve a more competitive production cost for surfactin production. The authors demonstrate the feasibility of improving surfactin production by modification of gene regulatory elements to direct the flow of the two peptides ComX pheromone and CSF and by using synthetic wastewater as a substrate. These approaches will be practical once the process of surfactin production becomes more economical and the production rates have met the commercial demands.

    19. Tissue Engineering and Delivery Systems

      Synergistic action of fibroblast growth factor-2 and transforming growth factor-beta1 enhances bioprinted human neocartilage formation (pages 2357–2368)

      Xiaofeng Cui, Kurt Breitenkamp, Martin Lotz and Darryl D'Lima

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24488

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      The simultaneous inkjet printing and photopolymerization of living human chondrocytes in a polymerizable PEG matrix gives three-dimensional structures that grow into well-anchored viable cartilage tissue. With proper growth factor stimulations the printed neocartialge demonstrated enhanced chondrogeneic phenotype as from implantation with high cell density while maintaining precise printing resolution which can only be achieved from lower cell density bioink.

    20. Structured three-dimensional co-culture of mesenchymal stem cells with meniscus cells promotes meniscal phenotype without hypertrophy (pages 2369–2380)

      Xiaofeng Cui, Akihiko Hasegawa, Martin Lotz and Darryl D'Lima

      Version of Record online: 22 MAR 2012 | DOI: 10.1002/bit.24495

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      3D structural co-culture of mesenchymal stem cells (MSC) and meniscal cells (MC) is attractive to engineer meniscus tissue. MSC as a trophic mediator promoted a robust meniscal phenotype without hypertrophy, with respect to fibrous collagen production and meniscal gene expression.

    21. Human mesenchymal stem cell position within scaffolds influences cell fate during dynamic culture (pages 2381–2391)

      Andrew B. Yeatts, Elyse M. Geibel, Fayola F. Fears and John P. Fisher

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24497

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      Human mesenchymal stem cells cultured in a tubular perfusion system bioreactor respond to culture conditions depending on location of the cells within three-dimensional scaffolds. These findings could give tissue engineers the ability to direct cellular outcome via culture conditions and cell location.

    22. Poly(ethylene glycol) cross-linked hemoglobin with antioxidant enzymes protects pancreatic islets from hypoxic and free radical stress and extends islet functionality (pages 2392–2401)

      Venkatareddy Nadithe, Deepa Mishra and You Han Bae

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24501

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      The objective of this study was to investigate the efficiency of PEG-based hemoglobin conjugates crosslinked with antioxidant enzymes for their ability to protect an oxygen carrier (hemoglobin) and insulin secreting islets from the combination of hypoxic and free radical stress under simulated transplantation conditions. The authors suggested that antioxidant enzymes play a significant role in hemoglobin protection and thus extended cell protection.

    23. Adenoviral transduction supports matrix expression of alginate cultured articular chondrocytes (pages 2402–2408)

      D. Pohle, R. Kasch, P. Herlyn, R. Bader, T. Mittlmeier, B.M. Pützer and B. Müller-Hilke

      Version of Record online: 17 APR 2012 | DOI: 10.1002/bit.24505

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      Adenoviral transduced human articular chondrocytes express the transgene GFP for up to 13 weeks. After expansion in monolayer and subsequent culture in 3D in alginate the transduced chondrocytes produce not only more proteoglycans, but also display an increased anabolic and decreased catabolic activity when compared to non-transduced controls.

  6. Communication to the Editor

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Editors' Choice
    6. Articles
    7. Communication to the Editor
    1. Cellular and Metabolic Engineering

      Increased resistance to oxysterol cytotoxicity in fibroblasts transfected with a lysosomally targeted Chromobacterium oxidase (pages 2409–2415)

      Jacques M. Mathieu, Fan Wang, Laura Segatori and Pedro J. Alvarez

      Version of Record online: 8 APR 2012 | DOI: 10.1002/bit.24506

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      Wild type human fibroblasts were transfected with a lysosomally targeted oxidase. The EGFP-tagged protein is shown here colocalized with acridine orange (bottom left). Transfected cells experienced morphological changes as compared to control (bottom right), but displayed decreased sensitivity towards 7-ketocholesterol cytotoxicity.

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