Biotechnology and Bioengineering

Cover image for Vol. 111 Issue 2

February 2014

Volume 111, Issue 2

Pages C1–C1, fmi–fmv, vi–vi, 209–424

  1. Cover

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

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communications to the Editor
    1. You have free access to this content
  3. Features

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communications to the Editor
    1. Spotlight

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      Matrix influences drug response (page vi)

      Version of Record online: 19 DEC 2013 | DOI: 10.1002/bit.25033

  4. Review

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communications to the Editor
    1. Materials-based strategies for multi-enzyme immobilization and co-localization: A review (pages 209–222)

      Feng Jia, Balaji Narasimhan and Surya Mallapragada

      Version of Record online: 14 NOV 2013 | DOI: 10.1002/bit.25136

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      Materials-based strategies for multi-enzyme immobilization and co-localization

  5. Articles

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communications to the Editor
    1. Biocatalysis, Protein Engineering, and Nanobiotechnology

      Coupling dark metabolism to electricity generation using photosynthetic cocultures (pages 223–231)

      Jonathan P. Badalamenti, César I. Torres and Rosa Krajmalnik-Brown

      Version of Record online: 14 AUG 2013 | DOI: 10.1002/bit.25011

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      Photosynthetic microbial electrochemical cells (MXCs) using cocultures of Geobacter and Chlorobium generated electricity with a negative light response similar to that observed in previous phototrophic enrichments. Current production increased in the dark and coincided with fluctuations in glycogen content, suggesting that dark fermentation yielded a metabolite available for anode respiration by Geobacter. When electrodes were not present, Geobacter was dependent on Chlorobium for its electron donor and acceptor via temporal separation of photosynthesis and dark fermentation.

    2. A high-throughput screen for antibiotic drug discovery (pages 232–243)

      Thomas C. Scanlon, Sarah M. Dostal and Karl E. Griswold

      Version of Record online: 29 AUG 2013 | DOI: 10.1002/bit.25019

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      Ultra-high-throughput screen for antibiotic drug discovery. A and B: Environmental DNA is cloned into a vector for heterologous expression in E. coli hosts. C: A microfluidic device is used to generate agarose-in-oil micro-emulsions in which individual recombinant E. coli are co-encapsulated with live bacterial pathogens. D: Droplets in which the recombinant clone secretes a biologic antibiotic are labeled with the SyTox Orange viability probe (orange circles). E: Droplet populations are sorted by FACS to isolate recombinant hosts secreting bactericidal natural products.

    3. Extension and application of the “enzyme test bench” for oxygen consuming enzyme reactions (pages 244–253)

      Kirill Rachinskiy, Martin Kunze, Careen Graf, Hergen Schultze, Matthias Boy and Jochen Büchs

      Version of Record online: 11 SEP 2013 | DOI: 10.1002/bit.25020

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      Within process development, powerful screening techniques are required to select the optimal biocatalyst. One of the most important characteristics for industrial application is the enzyme's long-term stability. To address this issue, a new model based technique called “enzyme test bench” is presented, which combines the high throughput screening approach with an extensive enzyme characterization. In this work the “enzyme test bench” technique was extended and applied to characterize and optimize a laccase-mediator system as representative for oxygen consuming enzyme reactions.

    4. Biofuels and Environmental Biotechnology

      Severity factor coefficients for subcritical liquid hot water pretreatment of hardwood chips (pages 254–263)

      Youngmi Kim, Thomas Kreke, Nathan S. Mosier and Michael R. Ladisch

      Version of Record online: 22 AUG 2013 | DOI: 10.1002/bit.25009

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      Glucose yields from enzyme hydrolysis of liquid hot water pretreated hardwood fall on a single, characteristic curve when the constant ω = 4.6 is used to correlate pretreatment severity to hydrolysis (Figure 4F). When the previously published value of ω = 14.75 is employed, four distinct curves result (Figure 4E). This work shows a simple adjustment of the value of ω enables correlation of glucose yield at pretreatment temperatures of 140 to 230°C.

    5. Enzymatic hydrolysis of pelletized AFEX™-treated corn stover at high solid loadings (pages 264–271)

      Bryan D. Bals, Christa Gunawan, Janette Moore, Farzaneh Teymouri and Bruce E. Dale

      Version of Record online: 20 SEP 2013 | DOI: 10.1002/bit.25022

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      AFEX is a unique pretreatment in that it can be performed at small depots followed by pelletization and shipping to large biorefineries. The authors have demonstrated that these pellets are effectively saccharified at high solid loading despite a decrease in water retention. Due to the low water retention, mixing during high solid hydrolysis is greatly improved.

    6. Dynamic model-based analysis of furfural and HMF detoxification by pure and mixed batch cultures of S. cerevisiae and S. stipitis (pages 272–284)

      Timothy J. Hanly and Michael A. Henson

      Version of Record online: 7 OCT 2013 | DOI: 10.1002/bit.25101

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      In this study, the authors incorporate the effects of furfural and HMF on pure and mixed yeast cultures of Saccharomyces cerevisiae and Scheffersomyces stipitis in dynamic flux balance analysis. The model was used to determine the process parameters that maximize co-culture ethanol production from sugar mixtures containing these inhibitors that are found in many lignocellulosic hydrolysates.

    7. Mass transfer studies of Geobacter sulfurreducens biofilms on rotating disk electrodes (pages 285–294)

      Jerome T. Babauta and Haluk Beyenal

      Version of Record online: 19 SEP 2013 | DOI: 10.1002/bit.25105

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      Geobacter sulfurreducens forms conductive biofilms that grow on electrode surfaces and transport electrons through extracellular electron transfer pathways. In this work, a rotating disk electrode was used to demonstrate the effect of mass transfer processes on electron transfer by G. sulfurreducens biofilms using electrochemical impedance spectroscopy. Under starving conditions, the pseudocapacitive nature of the biofilm impedance was readily observed and suggests that electron donor limitations can be identified by an additional capacitive element in the biofilm equivalent electrical circuit model.

    8. Bioprocess Engineering and Supporting Technologies

      Liquid films on shake flask walls explain increasing maximum oxygen transfer capacities with elevating viscosity (pages 295–308)

      Heiner Giese, Amizon Azizan, Anne Kümmel, Anping Liao, Cyril P. Peter, João A. Fonseca, Robert Hermann, Tiago M. Duarte and Jochen Büchs

      Version of Record online: 29 AUG 2013 | DOI: 10.1002/bit.25015

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      Oxygen concentration profiles in liquid films on the wall of rotating shake flasks were numerically simulated. Higbie's film theory overestimates oxygen flux into liquid films for thin films and low shaking frequencies. It was experimentally shown that oxygen transfer capacity (OTRmax) increases with increasing viscosities up to 10 mPa s, resulting in an increasing oxygen supply for microorganisms. At elevated viscosities of up to 80 mPa s, the OTRmax remains higher than the OTRmax at waterlike viscosities – in contrast to stirred tanks.

    9. A model to assess the feasibility of shifting reaction equilibrium by acetone removal in the transamination of ketones using 2-propylamine (pages 309–319)

      Pär Tufvesson, Christian Bach and John M. Woodley

      Version of Record online: 26 AUG 2013 | DOI: 10.1002/bit.25017

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      In this work, experimental data for acetone removal from water was fitted to a model. This model was subsequently coupled to a model for a transaminase biocatalyst and used to simulate processes with different target amines using 2-propylamine as amine donor. Based on the reaction equilibrium (Keq) of the transamination and the volatility of the starting material the model can be used to assess the feasibility of an investigated process before an engineered biocatalyst is available.

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      Mechanistic modeling of sulfur-deprived photosynthesis and hydrogen production in suspensions of Chlamydomonas reinhardtii (pages 320–335)

      C. R. Williams and M.A. Bees

      Version of Record online: 11 SEP 2013 | DOI: 10.1002/bit.25023

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      A novel mechanistic model for sulphur-deprived hydrogen gas production is developed from a consideration of the essential pathways and processes, including feedback between sulfur uptake, photosynthetic growth and endogenous substrate. Good agreement between model results and published experimental studies for trends in hydrogen yield and initiation time is found, and the model is employed to probe optimal external sulfur and illumination conditions. The model constitutes a powerful theoretical tool for investigating how hydrogen production from algae can be optimised.

    11. Cellular and Metabolic Engineering

      Molecular farming in tobacco hairy roots by triggering the secretion of a pharmaceutical antibody (pages 336–346)

      Suvi T. Häkkinen, Nicole Raven, Maurice Henquet, Marja-Leena Laukkanen, Tibor Anderlei, Juha-Pekka Pitkänen, Richard M. Twyman, Dirk Bosch, Kirsi-Marja Oksman-Caldentey, Stefan Schillberg and Anneli Ritala

      Version of Record online: 24 SEP 2013 | DOI: 10.1002/bit.25113

    12. Engineering of Saccharomyces cerevisiae for the synthesis of short chain fatty acids (pages 347–358)

      Christopher Leber and Nancy A. Da Silva

      Version of Record online: 3 SEP 2013 | DOI: 10.1002/bit.25021

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      Saccharomyces cerevisiae was engineered to produce short chain fatty acids (SCFA: C6 to C10) from glucose using the heterologous Homo sapiens type I fatty acid synthase (hFAS), activated by bacterial phosphopantetheine transferases (PPT). Short-chain thioesterases (TEs) were maintained on plasmids or linked to the hFAS, replacing the native TE domain. Overexpression of the linked hFAS variants increased caprylic (C8) acid production up to 64-fold (63 mg/L) over native yeast levels. Combined over-expression of the PPT led to C8 titers of up to 82 mg/L and total SCFA titers of up to 111 mg/L.

    13. Systems Biotechnology

      Beyond growth rate 0.6: What drives Corynebacterium glutamicum to higher growth rates in defined medium (pages 359–371)

      Simon Unthan, Alexander Grünberger, Jan van Ooyen, Jochem Gätgens, Johanna Heinrich, Nicole Paczia, Wolfgang Wiechert, Dietrich Kohlheyer and Stephan Noack

      Version of Record online: 24 SEP 2013 | DOI: 10.1002/bit.25103

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      The amino acid producer Corynebacterium glutamicum was cultivated in bioreactors in well-known CGXII medium at low inoculum (OD ≈ 0.005). Interestingly, in this highly diluted environment a growth rate of was observed, reproducing the authors' former results from microfluidic single cell cultivations (MSCC). Advanced growth studies, combining bioreactor and MSCC experiments, revealed a hidden co-substrate within CGXII. Protocatechuic acid (PCA), originally classified as iron chelator, is co-metabolized next to glucose and significantly accelerates growth by direct TCA feed.

    14. Model-directed engineering of “difficult-to-express” monoclonal antibody production by Chinese hamster ovary cells (pages 372–385)

      Leon P. Pybus, Greg Dean, Nathan R. West, Andrew Smith, Olalekan Daramola, Ray Field, Stephen J. Wilkinson and David C. James

      Version of Record online: 14 NOV 2013 | DOI: 10.1002/bit.25116

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      “Difficult-to-express” (DTE) proteins are becoming increasingly abundant in the development pipelines of many biopharmaceutical companies. The low process titers obtained whilst expressing such molecules typically delay or block their route to market. In this study the authors utilise a panel of recombinant monoclonal antibodies (MAb) that varied markedly in their “expressability” and employ detailed empirical based modelling of their cellular synthesis and secretion pathway to rationally design and implement engineering strategies to increase DTE MAb production.

    15. Tissue Engineering and Delivery Systems

      Biomimetic construction of cellular shell by adjusting the interfacial energy (pages 386–395)

      Ben Wang, Peng Liu, Zhaoming Liu, Haihua Pan, Xurong Xu and Ruikang Tang

      Version of Record online: 22 AUG 2013 | DOI: 10.1002/bit.25016

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      Inspired by nature, cytocompatible methods for cells encapsulated inside a mineral shell were developed by adjusting the interfacial energy and the precipitation of calcium minerals can be induced on the yeast cell surfaces, however, maintaining cell viability. These biomineralization treatments of single-cell, or so-called “cellular shellization”, may be applied as a potential and universal approach for cell-based sensing and therapy.

    16. Multiwell stiffness assay for the study of cell responsiveness to cytotoxic drugs (pages 396–403)

      Silviya Zustiak, Ralph Nossal and Dan L. Sackett

      Version of Record online: 6 SEP 2013 | DOI: 10.1002/bit.25097

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      A novel multiwell stiffness assay based on collagen-coated polyacrylamide gel was developed. The stiffness assay, due to its multiwell plate format, could be an excellent tool for studying (a) stiffness-dependent cell fate as well as (b) drug responses. The neuroblastoma cell line SY5Y is given as an example.

    17. Automated, spatio-temporally controlled cell microprinting with polymeric aqueous biphasic system (pages 404–412)

      David Petrak, Ehsan Atefi, Liya Yin, William Chilian and Hossein Tavana

      Version of Record online: 11 SEP 2013 | DOI: 10.1002/bit.25100

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      A fully-automated, cell microprinting technology using a polymeric aqueous two-phase system (ATPS) was developed. This approach enables non-contact printing of cells onto any substrate including a cell layer and decellularized films with spatial and temporal control over positioning of cells in user-defined shapes. This technology will be useful for tissue engineering applications to construct organized cell layers composed of multiple cell types.

  6. Communications to the Editor

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communications to the Editor
    1. Bioseparations and Downstream Processing

      Novel thermo-responsive fucose binding ligands for glycoprotein purification by affinity precipitation (pages 413–417)

      Lindsay Arnold and Rachel Chen

      Version of Record online: 8 OCT 2013 | DOI: 10.1002/bit.25118

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      Designed and developed thermo-responsive fucose-binding lectins. Successfully demonstrated their use in non-chromatographic glycoprotein purification. The new method affords high yield (complete recovery of glycoprotein) and high sensitivity (applicable to protein concentration as low as 1.4 pm) and effective in the presence of contaminants.

    2. Tissue Engineering and Delivery Systems

      Drug susceptibility of matrix-encapsulated Candida albicans nano-biofilms (pages 418–424)

      Anand Srinivasan, Celia Macias Gupta, C. Mauli Agrawal, Kai P. Leung, Jose L. Lopez-Ribot and Anand K. Ramasubramanian

      Version of Record online: 18 OCT 2013 | DOI: 10.1002/bit.25120

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      Nano-scale cultures of Candida albicans biofilms encapsulated in collagen or alginate matrices in a microarray were used to test the susceptibility of antifungal drugs. Srinivasan et al show that these cultures exhibit differential response to drugs depending upon the interaction between the drug and the matrix. Their results underscore the choice of matrix as a critical parameter in 3D cell encapsulation for drug discovery and tissue engineering applications.

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