Biotechnology and Bioengineering

Cover image for Vol. 111 Issue 4

April 2014

Volume 111, Issue 4

Pages C1–C1, fmi–fmv, vi–vi, 639–847

  1. Cover

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

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

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  4. Review

    1. Top of page
    2. Cover
    3. Contents
    4. Features
    5. Review
    6. Articles
    7. Communication to the Editor
    1. Lipase-catalyzed process for biodiesel production: Protein engineering and lipase production (pages 639–653)

      Hyun Tae Hwang, Feng Qi, Chongli Yuan, Xuebing Zhao, Doraiswami Ramkrishna, Dehua Liu and Arvind Varma

      Article first published online: 17 DEC 2013 | DOI: 10.1002/bit.25162

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      This review provides an overview of the recent advances in lipase engineering, including both protein modification and production. Recent advances in biotechnology such as in protein engineering, recombinant methods and metabolic engineering have been employed but are yet to impact lipase engineering for cost-effective production of biodiesel. A summary of the current challenges and perspectives for potential solutions are also provided.

  5. Articles

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

      Enzyme-driven bacillus spore coat degradation leading to spore killing (pages 654–663)

      Ruchir V. Mundra, Krunal K. Mehta, Xia Wu, Elena E. Paskaleva, Ravi S. Kane and Jonathan S. Dordick

      Article first published online: 6 NOV 2013 | DOI: 10.1002/bit.25132

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      The authors have developed an enzymatic route towards bacillus spore decontamination. Central to this outside in decontamination approach, is the action of proteases that catalyze partial degradation of the spore coat of B. cereus and B. anthracis, which leads to a morphological change in the otherwise impregnable coat structure, increasing coat permeability to various cortex and cell lytic enzymes. The elimination of detergents and strong denaturants reduces the environmental burden of chemically mediated spore killing, and demonstrates that a mild and environmentally benign biocatalytic spore killing is achievable.

    2. Cel48A from Thermobifida fusca: Structure and site directed mutagenesis of key residues (pages 664–673)

      Maxim Kostylev, Markus Alahuhta, Mo Chen, Roman Brunecky, Michael E. Himmel, Vladimir V. Lunin, John Brady and David B. Wilson

      Article first published online: 21 NOV 2013 | DOI: 10.1002/bit.25139

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      In this work the authors present a structure of TfCel48A, a family 48 cellulase from a model cellulolytic bacterium Thermobifida fusca. Site-directed mutagenesis combined with time course assays identifies key aromatic residues that are important for the ability of TfCel48a to digest crystalline and amorphous cellulosic substrates.

    3. Deletion of a dynamic surface loop improves stability and changes kinetic behavior of phosphatidylinositol-synthesizing Streptomyces phospholipase D (pages 674–682)

      Jasmina Damnjanović, Hideo Nakano and Yugo Iwasaki

      Article first published online: 30 NOV 2013 | DOI: 10.1002/bit.25149

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      Thermostabilization of a phosphatidylinositol (PI)-synthesizing phospholipase D was achieved by rational design. Deletion of an unstable, nine-residue loop resulted in a variant with 11.7 times longer activity half-life at 70°C than that of the parent enzyme. The stabilized variant enabled high-temperature PI synthesis from phosphatidylcholine and myo-inositol, providing 2-fold higher PI and lower side-product, phosphatidic acid yields.

    4. SunnyTALEN: A second-generation TALEN system for human genome editing (pages 683–691)

      Ning Sun, Zehua Bao, Xiong Xiong and Huimin Zhao

      Article first published online: 11 DEC 2013 | DOI: 10.1002/bit.25154

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      In this study, the authors used directed evolution to create a TALEN variant with enhanced activity, SunnyTALEN. The corresponding scaffold increases the rate of genetic modification at all the 13 tested loci of human genome and is compatible with heterodimer TALEN architectures. This high-efficiency TALEN variant represents a novel second-generation TALEN system and has great potential for biological and therapeutic applications.

    5. Biofuels and Environmental Biotechnology

      Oxygen allows Shewanella oneidensis MR-1 to overcome mediator washout in a continuously fed bioelectrochemical system (pages 692–699)

      Michaela A. TerAvest, Miriam A. Rosenbaum, Nicholas J. Kotloski, Jeffrey A. Gralnick and Largus T. Angenent

      Article first published online: 29 OCT 2013 | DOI: 10.1002/bit.25128

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      The authors investigated the importance of flavins—endogenous electron mediators—in electron transfer between Shewanella oneidensis MR-1 and anode electrodes under anaerobic and micro-aerobic conditions. Micro-aerobic conditions promoted mediated electron transfer via flavins under continuously fed conditions, but had little effect on the electron transfer mechanism under batch-fed conditions. This indicates that oxygen allowed S. oneidensis to overcome flavin washout in the continuously fed condition, either by increasing cell mass or by increasing per cell flavin production. This result may have broader implications for both mixed- and pure-culture bioelectrochemical systems harboring aerotolerant organisms.

    6. Enhancing E. coli isobutanol tolerance through engineering its global transcription factor cAMP receptor protein (CRP) (pages 700–708)

      Huiqing Chong, Hefang Geng, Hongfang Zhang, Hao Song, Lei Huang and Rongrong Jiang

      Article first published online: 6 NOV 2013 | DOI: 10.1002/bit.25134

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      Error-prone PCR was employed to engineer global regulator cAMP receptor protein (CRP) of E. coli to enhance its isobutanol tolerance. Variant IB2 (S179P, H199R) was isolated from the random mutagenesis library, which exhibited much better growth (0.18 h-1) in 1.2% (v/v) isobutanol (9.6 g/l), much better than the control (0.05 h-1) and crp knock out strain.

    7. A defined co-culture of Geobacter sulfurreducens and Escherichia coli in a membrane-less microbial fuel cell (pages 709–718)

      Nicholas Bourdakos, Enrico Marsili and Radhakrishnan Mahadevan

      Article first published online: 22 NOV 2013 | DOI: 10.1002/bit.25137

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      Co-culture microbial fuel cell (MFC) with E. coli and G. sulfurreducens fed with glucose-rich medium produce sustainable power higher that that measured in a single culture E. coli MFC.

    8. Nanoscale interactions of polyethylene glycol with thermo-mechanically pre-treated Pinus radiata biofuel substrate (pages 719–725)

      Lloyd A. Donaldson, Roger H. Newman and Alankar Vaidya

      Article first published online: 18 NOV 2013 | DOI: 10.1002/bit.25138

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      PEG is a useful additive to enzymatic saccharification of pre-treated softwood biomass which improves glucose yield by reducing non-specific binding of enzyme to lignin. The authors have used confocal fluorescence microscopy combined with Förster resonance energy transfer (FRET) to reveal molecular level interactions between lignin present in thermo-mechanically pre-treated Pinus radiata substrate, and fluorescently labeled PEG.

    9. Cellulose hydrolysis and binding with Trichoderma reesei Cel5A and Cel7A and their core domains in ionic liquid solutions (pages 726–733)

      Ronny Wahlström, Jenni Rahikainen, Kristiina Kruus and Anna Suurnäkki

      Article first published online: 20 NOV 2013 | DOI: 10.1002/bit.25144

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      Ionic liquids are interesting media for biomass pretreatment, but they inactivate cellulases. In this work, the effect of two cellulose-dissolving ionic liquids on cellulose hydrolysis and substrate binding was studied with Trichoderma reesei endoglucanase Cel5A and cellobiohydrolase Cel7A and their respective core domains. According to the results, cellulase binding is severely interfered by the presence of ionic liquids, but to different degrees depending on the cellulase structure and mode of function.

    10. Bioprocess Engineering and Supporting Technologies

      Model-based derivation, analysis and control of unstable microaerobic steady-states—Considering Rhodospirillum rubrum as an example (pages 734–747)

      Lisa Carius, Philipp Rumschinski, Timm Faulwasser, Dietrich Flockerzi, Hartmut Grammel and Rolf Findeisen

      Article first published online: 27 NOV 2013 | DOI: 10.1002/bit.25140

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      Setup of the automatized continuous cultivation system. The redox potential of the culture broth was measured and controlled as described previosuly [Carius et al., 2013] (red shaded area). The infrastructure of the model-based 2DOF control strategy, which employs the online monitored biomass concentration as feedback signal, is shown in the blue-shaded area. The green-shaded area depicts the control loop of the harvesting, which the authors employ to keep the liquid volume of the culture constant.

    11. Cell line profiling to improve monoclonal antibody production (pages 748–760)

      Sohye Kang, Da Ren, Gang Xiao, Kristi Daris, Lynette Buck, Atim A. Enyenihi, Roman Zubarev, Pavel V. Bondarenko and Rohini Deshpande

      Article first published online: 19 NOV 2013 | DOI: 10.1002/bit.25141

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      Inherent, genetic features of various monoclonal antibody-producing CHO cell lines were examined using microarray-based transcriptomics and LC-MS/MS shotgun proteomics approaches. Specific transcripts and proteins correlating with productivity, proliferation rate and cell size have been identified. Global structures of the expression profiles were also analyzed and compared in order to assess their contribution toward specific culture phenotype.

    12. Cellular and Metabolic Engineering

      Enhanced production of coenzyme Q10 by self-regulating the engineered MEP pathway in Rhodobacter sphaeroides (pages 761–769)

      Wenqiang Lu, Lidan Ye, Haoming Xu, Wenping Xie, Jiali Gu and Hongwei Yu

      Article first published online: 31 OCT 2013 | DOI: 10.1002/bit.25130

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      A self-regulating system has been developed to control the expression level of the engineered pathway via the co-expressed LacIq protein. This system could flexibly regulate the engineered pathway to balance with the host metabolic pathways. With this system, the MEP pathway in Rhodobacter sphaeroides was optimized and the production of coenzyme Q10 was improved two-folds.

    13. Exploring the transcriptome space of a recombinant BHK cell line through next generation sequencing (pages 770–781)

      Kathryn C. Johnson, Andrew Yongky, Nandita Vishwanathan, Nitya M. Jacob, Karthik P. Jayapal, Chetan T. Goudar, George Karypis and Wei-Shou Hu

      Article first published online: 19 NOV 2013 | DOI: 10.1002/bit.25135

    14. Engineering Science of Biological Systems

      Aggregates of α-chymotrypsinogen anneal to access more stable states (pages 782–791)

      Ronald W. Maurer, Alan K. Hunter, Anne S. Robinson and Christopher J. Roberts

      Article first published online: 18 NOV 2013 | DOI: 10.1002/bit.25129

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    15. Deceleration-stats save much time during phototrophic culture optimization (pages 792–802)

      Sebastiaan Hoekema, Arjen Rinzema, Johannes Tramper, René H. Wijffels and Marcel Janssen

      Article first published online: 26 NOV 2013 | DOI: 10.1002/bit.25131

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      Model simulations show that the Deceleration stat (D-stat) optimization method is an attractive alternative to a series of chemostats in order to assess the maximal biomass production rate of microalgae photobioreactors. In addition, also the accurate determination of the corresponding microalgae specific growth rate is possible with significant time gain.

    16. Systems Biotechnology

      A model-driven quantitative metabolomics analysis of aerobic and anaerobic metabolism in E. coli K-12 MG1655 that is biochemically and thermodynamically consistent (pages 803–815)

      Douglas McCloskey, Jon A. Gangoiti, Zachary A. King, Robert K. Naviaux, Bruce A. Barshop, Bernhard O. Palsson and Adam M. Feist

      Article first published online: 19 NOV 2013 | DOI: 10.1002/bit.25133

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      Using quantitative metabolomics and a novel rapid sampling apparatus that was validated for aerobic and anaerobic growth, the authors provide a snapshot of the differences between aerobic and anaerobic steady-state metabolomes. Subsequent statistical and thermodynamic analysis highlighted several important biochemical features including the promiscuity of the gene product of pykA under anaerobic growth.

    17. Metabolic profiling of insect cell lines: Unveiling cell line determinants behind system's productivity (pages 816–828)

      Francisca Monteiro, Vicente Bernal, Xavier Saelens, Ana B. Lozano, Cristina Bernal, Angel Sevilla, Manuel J.T. Carrondo and Paula M. Alves

      Article first published online: 20 NOV 2013 | DOI: 10.1002/bit.25142

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      Sf9 and High Five cell lines have significantly different productivities regarding virus and recombinant protein production; therefore their metabolic profile was subjected to cross-comparison in order to identify traits leveraging productivity and specific pathways of paramount importance to support it. The most relevant pathways involved in baculovirus infection were identified and correlated with recombinant protein production. In blue are depicted the pathways highlighted in both Sf9 and High Five cells and in orange pathways recruited only in High Five cells.

    18. Tissue Engineering and Delivery Systems

      The potential of encapsulating “raw materials” in 3D osteochondral gradient scaffolds (pages 829–841)

      Neethu Mohan, Vineet Gupta, BanuPriya Sridharan, Amanda Sutherland and Michael S. Detamore

      Article first published online: 30 NOV 2013 | DOI: 10.1002/bit.25145

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      Raw material encapsulated microspheres were used to fabricate 3D gradient scaffolds for osteochondral regeneration. Chondroitin sulfate and bioactive glass served as raw materials in the microspheres and enabled superior performance of tissue engineered constructs. They are promising candidates that can be used to either replace or be used in combination with growth factors for more rapid translation of regenerative medicine products to the clinic.

  6. Communication to the Editor

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

      The importance of pyroglutamate in cellulase Cel7A (pages 842–847)

      Craig M. Dana, Alexandra Dotson-Fagerstrom, Christine M. Roche, Sarala M. Kal, Harshal A. Chokhawala, Harvey W. Blanch and Douglas S. Clark

      Article first published online: 28 JAN 2014 | DOI: 10.1002/bit.25178

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      The authors discovered that recombinant Cel7A expressed in Saccharomyces cerevisiae can lack cyclization of its N-terminal glutamine to pyroglutamate, leaving the N-terminal residue as regular glutamine. Relative to Cel7A expressed in the filamentous fungus Neurospora crassa, the uncyclized version has a lower Tm (by 10°C) and half the specific activity at 65°C. Treating the uncyclized enzyme in vitro with glutaminyl cyclase increased these properties to match those of Cel7A expressed in N. crassa.

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