Biotechnology and Bioengineering

Cover image for Biotechnology and Bioengineering

April 2013

Volume 110, Issue 4

Pages C1–C1, fmi–fmvi, 1005–1254

  1. Cover

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communication to the Editors
    1. You have free access to this content
      Biotechnology and Bioengineering: Volume 110, Number 4, April 2013 (page C1)

      Version of Record online: 22 FEB 2013 | DOI: 10.1002/bit.24656

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      Cover Legend Mixing visualization experiments of a 30 mL eccentric stirred tank mini-bioreactor. (Image courtesy of Dr. Mario M. Alvarez et. al.).

  2. Contents

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

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

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communication to the Editors
    1. Review

      You have full text access to this OnlineOpen article
      Oxygen transfer characteristics of miniaturized bioreactor systems (pages 1005–1019)

      Timothy V. Kirk and Nicolas Szita

      Version of Record online: 17 JAN 2013 | DOI: 10.1002/bit.24824

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      Dissolved oxygen (DO) transfer performance of submilliliter microbioreactors, and 1–10 mL minibioreactors was examined. Further, mathematical relationships confirming the applicability of kLa measurements across all scales have been derived. Finally, the influence on reactor performance of oxygen uptake rate (OUR), and the possibility of its real-time measurement have been explored.

  5. Articles

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

      Light-responsive current generation by phototrophically enriched anode biofilms dominated by green sulfur bacteria (pages 1020–1027)

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

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24779

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      Photosynthetic bacteria can incorporate light energy and/or carbon into microbial processes occurring in microbial electrochemical cells (MXCs). Following a stepwise selective enrichment strategy for anoxygenic phototrophs that could act as anode-respiring bacteria (ARB), the authors describe light-responsive current generation by electrode-associated biofilms whereby current decreased upon illumination. Green sulfur bacteria were predominant in fresh- and saltwater enrichments, and the authors propose that these phototrophs were responsible for both the negative light-responsiveness and novel anode respiration patterns observed by cyclic voltammetry.

    2. Biocatalysis, Protein Engineering, and Nanobiotechnology

      Engineering hyperthermostability into a mesophilic family 11 xylanase from Aspergillus oryzae by in silico design of N-terminus substitution (pages 1028–1038)

      Shu-Juan Gao, Jun-Qing Wang, Min-Chen Wu, Hui-Min Zhang, Xin Yin and Jian-Fang Li

      Version of Record online: 1 NOV 2012 | DOI: 10.1002/bit.24768

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      The N-terminus of AoXyn11A to be replaced by the corresponding region of EvXyn11TS was selected by comparing B-factors between AoXyn11A and EvXyn11TS, and confirmed by comparing the total energy value of AEx11A with that of AoXyn11A. Based on the in silico design of N-terminus substitution, the AEx11A was constructed and expressed in P. pastoris. The temperature optimum of reAEx11A was 30°C higher than that of reAoXyn11A, and its half-life was 197-fold longer than that of reAoXyn11A at 70°C.

    3. Biofuels and Environmental Biotechnology

      Co-hydrolysis of lignocellulosic biomass for microbial lipid accumulation (pages 1039–1049)

      Zhenhua Ruan, Michael Zanotti, Yuan Zhong, Wei Liao, Chad Ducey and Yan Liu

      Version of Record online: 26 NOV 2012 | DOI: 10.1002/bit.24773

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      Mass balance analysis demonstrated the effects of co-hydrolysates on utilization efficiency of different lignocellulosic biomass for microbial lipid production. It indicated that the oleaginous fungus M. isabellina lipid accumulation on co-hydrolysates was much superior to other widely studied lipid producing strains such as yeasts. Hence, oleaginous fungus could be a better microbial option to accumulate lipid from lignocellulosic biomass.

    4. Alternation in colonization behaviors of Escherichia coli cells with rpoS or yggE deficiency on solid surfaces (pages 1050–1056)

      Minh Hong Nguyen, Yoshihiro Ojima, Teruyoshi Kawata and Masahito Taya

      Version of Record online: 17 JAN 2013 | DOI: 10.1002/bit.24770

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      The authors clarified the contribution of flagella and curli to E. coli BW25113 colonization by using the yggE-deficient (ΔyggE) and rpoS-deficient (ΔrpoS) strains. Under a static condition, the plentiful flagella synthesized on ΔrpoS and ΔyggE cell surfaces facilitated the increased colonization. However, under a flow condition, only ΔyggE cells that formed abundant flagella and curli on their surfaces could maintain the stable colonization on the solid surface.

    5. Balancing redox cofactor generation and ATP synthesis: Key microaerobic responses in thermophilic fermentations (pages 1057–1065)

      Wesley Loftie-Eaton, Mark Taylor, Kerry Horne, Marla I. Tuffin, Stephanie G. Burton and Don A. Cowan

      Version of Record online: 25 DEC 2012 | DOI: 10.1002/bit.24774

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      Geobacillus thermoglucosidasius is an economically important organism for biofuel production yet little is known regarding its physiology. The authors measured specific transcriptional changes in the central metabolic pathways and global shifts in the proteome of this Gram-positive thermophile in response to a switch from aerobic respiration to microaerobic fermentation. Interesting results included the presence of a putative oxygen scavenging electron transport chain and a glyoxalase pathway in addition to a general response to low pO2.

    6. Biocatalytic reduction of short-chain carboxylic acids into their corresponding alcohols with syngas fermentation (pages 1066–1077)

      Jose M. Perez, Hanno Richter, Sarah E. Loftus and Largus T. Angenent

      Version of Record online: 15 JAN 2013 | DOI: 10.1002/bit.24786

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      The potential of carboxydotrophic bacteria for conversion of short chain carboxylic acids into alcohols, using syngas as a source of electrons and energy, is demonstrated. Acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid, and n-caproic acid were converted into their corresponding alcohols. An ATP yield of 0.4–0.5 mol of ATP per mol CO consumed was calculated in the presence of hydrogen. The ratio of protons pumped versus electrons transferred via the Rnf complex is suggested to be 1.0.

    7. Catalysis with CuII(bpy) improves alkaline hydrogen peroxide pretreatment (pages 1078–1086)

      Zhenglun Li, Charles H. Chen, Tongjun Liu, Vaidyanathan Mathrubootham, Eric L. Hegg and David B. Hodge

      Version of Record online: 29 DEC 2012 | DOI: 10.1002/bit.24793

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      Pretreatment with alkaline hydrogen peroxide (AHP) catalyzed by CuII(bpy) complex significantly improves enzymatic digestibility of woody biomass. Chemical modifications of plant cell wall components share features with in natura fungal decay.

    8. Bioprocess Engineering and Supporting Technologies

      Eliminating tyrosine sequence variants in CHO cell lines producing recombinant monoclonal antibodies (pages 1087–1097)

      Lauren Feeney, Veronica Carvalhal, X. Christopher Yu, Betty Chan, David A. Michels, Yajun Jennifer Wang, Amy Shen, Jan Ressl, Brendon Dusel and Michael W. Laird

      Version of Record online: 15 FEB 2013 | DOI: 10.1002/bit.24759

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      This work investigates a commonly observed sequence variant (SV) in several CHO cell produced monoclonal antibodies, and how the levels of key amino acids in the cell culture fermentation process can impact the level of SV. Specifically, several tyrosine residues throughout the protein were found to have either phenylalanine or histidine incorporated in place of tyrosine; it was determined that this event strongly correlated with tyrosine depletion. The data suggests that the mechanism of sequence variant formation is mRNA mistranslation.

    9. Polymer characterization and optimization of conditions for the enhanced bioproduction of benzaldehyde by Pichia pastoris in a two- ;phase partitioning bioreactor (pages 1098–1105)

      Tom Craig and Andrew J. Daugulis

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24778

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      Pichia pastoris has high alcohol oxidase activity and, although often directed at methanol metabolism, can non-specifically oxidize other alcohols, here benzyl alcohol to benzaldehyde, an important fragrance/flavour compound. Benzaldehyde strongly inhibited the biotransformation, however, and a Two-Phase Partitioning Bioreactor (TPPB) was employed using amorphous polymer beads to sequester the produced benzaldehyde. The uptake mechanism was confirmed to be by absorption, not adsorption, as is seen in non-selective materials such as activated carbon. The benzaldehyde volumetric productivity was double in the TPPB compared to single phase.

    10. A simple eccentric stirred tank mini-bioreactor: Mixing characterization and mammalian cell culture experiments (pages 1106–1118)

      David Bulnes-Abundis, Leydi M. Carrillo-Cocom, Diana Aráiz-Hernández, Alfonso García-Ulloa, Marisa Granados-Pastor, Pamela B. Sánchez-Arreola, Gayathree Murugappan and Mario M. Alvarez

      Version of Record online: 26 NOV 2012 | DOI: 10.1002/bit.24780

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      The authors study the mixing performance and mammalian cell culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Their results demonstrate good mixing performance. Mammalian cell culture experiments were conducted. Relevant biological indicators of CHO cell cultures grown in the mini-bioreactor were comparable to those observed in 1 L-fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded 10-fold using the eccentric system.

    11. Bioseparations and Downstream Processing

      Nanofiber adsorbents for high productivity downstream processing (pages 1119–1128)

      Oliver Hardick, Stewart Dods, Bob Stevens and Daniel G. Bracewell

      Version of Record online: 1 NOV 2012 | DOI: 10.1002/bit.24765

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      Electrospun polymeric nanofibers offer a possible solution to limited productivity in downstream processing. Here the authors fabricate cellulosic nanofibers functionalized with DEAE ligands. These are packed into a cartridge to promote flow distribution and convective mass transfer. The resultant material is resilient to fouling and can be operated at high flowrates while maintaining capacity. These excellent mass transfer properties offer significant productivity improvements (potentially 10 fold) if combined with continuous chromatography.

    12. You have full text access to this OnlineOpen article
      Zonal rate model for axial and radial flow membrane chromatography. Part I: Knowledge transfer across operating conditions and scales (pages 1129–1141)

      Pranay Ghosh, Kaveh Vahedipour, Min Lin, Jens H. Vogel, Charles A. Haynes and Eric von Lieres

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24771

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      The zonal rate model (ZRM) approach is extended from axial to radial flow configurations. Membrane chromatography capsules at different scales are analyzed under binding and non-binding conditions. The ZRM can help saving valuable material and time, as the experiments required for model calibration are much cheaper than the predicted large-scale experiments at binding conditions.

    13. The effect of protein a cycle number on the performance and lifetime of an anion exchange polishing step (pages 1142–1152)

      Timothy Iskra, Glen R. Bolton, Jonathan L. Coffman and Ranga Godavarti

      Version of Record online: 10 DEC 2012 | DOI: 10.1002/bit.24781

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      An understanding of the performance linkages between the unit operations can lead to robust manufacturing processes. In this study, a unique linkage between two chromatography unit operations is described. When material from a Protein A column that had been cycled a few times (<10) was loaded on the anion-exchange chromatography resin, premature loss of capacity was observed. Depth filtration of the Protein A peak pool was found to reduce the level of the foulant and significantly improve the AEX resin capacity, robustness and lifetime.

    14. Cellular and Metabolic Engineering

      IL-17F co- ;expression improves cell growth characteristics and enhances recombinant protein production during CHO cell line engineering (pages 1153–1163)

      Mathias Contie, Olivier Leger, Nicolas Fouque, Yves Poitevin, Marie Kosco-Vilbois, Nicolas Mermod and Greg Elson

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24763

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      This study demonstrates that IL-17F expression in CHO cells confers benefit during cell line engineering for recombinant protein production. Aside from enhanced cell line productivity, improvements in cell growth during selection and subcloning allowed for an overall time saving of up to one month during clonal cell line selection.

    15. Optimized signal peptides for the development of high expressing CHO cell lines (pages 1164–1173)

      Lars Kober, Christoph Zehe and Juergen Bode

      Version of Record online: 17 JAN 2013 | DOI: 10.1002/bit.24776

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      Highly productive cell lines are required for the production of biopharmaceutical products. To this end a variety of signal peptides was used to express different antibodies and non-antibody products in transiently and stably transfected Chinese hamster ovary (CHO) cells. Thereby it could be demonstrated that signal peptide B, but also signal peptide E can be used to generate cell lines with cell specific productivities (QP) up to 60 or 90 pg antibody/cell/day (pcd mean values are represented by red bars).

    16. You have full text access to this OnlineOpen article
      Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: Cytological studies (pages 1174–1179)

      Numfon Rakkhumkaew, Shigeo Shibatani, Takeru Kawasaki, Makoto Fujie and Takashi Yamada

      Version of Record online: 12 FEB 2013 | DOI: 10.1002/bit.24783

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      In this study, the authors describe the novel production of HA in cultured tobacco cells (BY-2). The tobacco cells were successfully transformed with the chloroviral hyaluronan synthase (cv-has) gene with or without a vacuolar targeting signal. The results showed that the sporamin vacuolar targeting signal (vSPO) operated well in BY-2 cells, targeting the cvHAS protein to the vacuolar membrane where HA was synthesized and transported into the storage vacuole.

    17. Engineering Science of Biological Systems

      Anaerobic ammonia oxidation (ANAMMOX) for side-stream treatment of anaerobic digester filtrate process performance and microbiology (pages 1180–1192)

      Shireen M. Kotay, Bryan L. Mansell, Mitch Hogsett, Huang Pei and Ramesh Goel

      Version of Record online: 12 NOV 2012 | DOI: 10.1002/bit.24767

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      Total nitrogen removal from the raw filtrate using ANAMMOX was described. Partial nitrification provided nitrite necessary for ANAMMOX. The Anammox reactor was dominated by K. stuttgartiensis while the partial nitritation reactor was dominated by Nitrosomonas europaea.

    18. Synthetic Biology

      Integrating cell-free biosyntheses of heme prosthetic group and apoenzyme for the synthesis of functional P450 monooxygenase (pages 1193–1200)

      Yong-Chan Kwon, In-Seok Oh, Nahum Lee, Kyung-Ho Lee, Yeo Joon Yoon, Eun Yeol Lee, Byung-Gee Kim and Dong-Myung Kim

      Version of Record online: 30 NOV 2012 | DOI: 10.1002/bit.24785

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      Functional holoenzyme of P450 monooxygenase was produced in a cell-free synthesis system. Step reactions required for the syntheses of apoenzyme and prosthetic group have been designed and implemented into a cell-free synthesis system so that these two separate pathways take place in the same reaction mixture, while being insulated from each other. Combined pathways for the synthesis of functional P450 monooxygenase were then further integrated with in situ assay reactions to enable real-time measurement of enzymatic activity during its synthesis.

    19. A yeast-based rapid prototype platform for gene control elements in mammalian cells (pages 1201–1210)

      Kathy Y. Wei, Yvonne Y. Chen and Christina D. Smolke

      Version of Record online: 17 JAN 2013 | DOI: 10.1002/bit.24792

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      Most existing gene-control systems are either mechanistically specific for microbial organisms or must be laboriously re-engineered to function in mammalian cells. The authors demonstrate a ribozyme-based device platform where devices prototyped in yeast to regulate gene expression can be directly transported to mammalian cells. Their unique device architecture acts as a rapid prototyping platform (RPP) based on a yeast chassis, providing a genetically tractable system that supports high-throughput screens for generating gene-controllers functional in mammalian cells.

    20. Tissue Engineering and Delivery Systems

      Targeted nitric oxide delivery preferentially induces glioma cell chemosensitivity via altered p53 and O6-Methylguanine-DNA Methyltransferase activity (pages 1211–1220)

      Shahana Safdar, Courtney A. Payne, Nam H. Tu and Lakeshia J. Taite

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24775

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      Glioma-specific NO donors were used to induced chemosensitivity in vitro. Cell viability of T98G and U-87MG malignant glioma cells is significantly decreased and levels of active MGMT and p53 are implicated in the mechanism of chemoresistance. Cell invasion is also decreased through the occupation of MMP-2 on glioma cell surfaces.

    21. In silico multi-scale model of transport and dynamic seeding in a bone tissue engineering perfusion bioreactor (pages 1221–1230)

      T.J. Spencer, L.A. Hidalgo-Bastida, S.H. Cartmell, I. Halliday and C.M. Care

      Version of Record online: 23 NOV 2012 | DOI: 10.1002/bit.24777

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      A mathematical model predicting the coupled fluid flow, mass transport and cell seeded attachment in a perfusion bioreactor for a bone tissue engineering application is developed and proposed. Model boundary conditions derive directly from microCT scans of the porous PLLA scaffold and bioreactor vessel enabling both small scale and large scale flow features with out resort to approximation. The models use for in silico design is thus demonstrated.

    22. Shear stress during early embryonic stem cell differentiation promotes hematopoietic and endothelial phenotypes (pages 1231–1242)

      Russell P. Wolfe and Tabassum Ahsan

      Version of Record online: 15 FEB 2013 | DOI: 10.1002/bit.24782

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      Application of shear stress during ESC differentiation in a 2D system is a robust method for promoting endothelial and hematopoietic phenotypes. Responses were similar for a range of stress magnitudes and different protein-coated surfaces. These effects, however, were dependent on the duration and time point of treatment exposure and required activation of the FLK1 protein. These results suggest that shear stress in microcarrier-based scalable systems may be one approach to generate the necessary number of cells needed for therapeutic applications.

  6. Communication to the Editors

    1. Top of page
    2. Cover
    3. Contents
    4. Spotlights
    5. Reviews
    6. Articles
    7. Communication to the Editors
    1. Bioprocess Engineering and Supporting Technologies

      Improved production of 3-hydroxypropionaldehyde by complex formation with bisulfite during biotransformation of glycerol (pages 1243–1248)

      Roya R.R. Sardari, Tarek Dishisha, Sang-Hyun Pyo and Rajni Hatti-Kaul

      Version of Record online: 10 DEC 2012 | DOI: 10.1002/bit.24787

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      3-Hydroxypropionaldehyde (3HPA) was produced from glycerol by Lactobacillus reuteri through a single enzyme catalyzed reaction. Complex formation with sodium bisulfite was studied and used for in situ capture of 3HPA in a fed-batch biotransformation, with prolonged bioactivity of the cells for at least 18 h and more than double yield of 3HPA than the process without bisulfite. The ratio of 3HPA to other products was also doubled.

    2. Tissue Engineering and Delivery Systems

      Fiber alignment directs cell motility over chemotactic gradients (pages 1249–1254)

      Harini G. Sundararaghavan, Randi L. Saunders, Daniel A. Hammer and Jason A. Burdick

      Version of Record online: 28 DEC 2012 | DOI: 10.1002/bit.24788

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      A major hurdle in utilizing tissue engineered scaffolds in vivo is the lack of cell infiltration and angiogenesis in implanted materials. The authors investigated competing influences of chemical factors and topographical cues (aligned fibers) on endothelial cell migration. Though chemical cues are the most common method used to direct cell migration, the authors found topographical cues are more influential, and overlaying these cues compounds their effects.

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