Biotechnology and Bioengineering

Cover image for Vol. 113 Issue 8

Early View (Online Version of Record published before inclusion in an issue)

Edited By: Douglas S. Clark

Impact Factor: 4.243

ISI Journal Citation Reports © Ranking: 2015: 24/161 (Biotechnology & Applied Microbiology)

Online ISSN: 1097-0290

  1. Articles

    1. Cell migration and organization in three-dimensional in vitro culture driven by stiffness gradient

      Danielle Joaquin, Michael Grigola, Gubeum Kwon, Christopher Blasius, Yutao Han, Daniel Perlitz, Jing Jiang, Yvonne Ziegler, Ann Nardulli and K. Jimmy Hsia

      Version of Record online: 3 JUN 2016 | DOI: 10.1002/bit.26010

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      Cells migrate in 3D culture with speed proportional to stiffness gradient.

  2. Communication To The Editor

    1. In vitro analysis of essential binding sites on the promoter of the Serratia marcescens spn operon with the quorum-sensing receptor SpnR

      Yuriko Takayama and Norihiro Kato

      Version of Record online: 3 JUN 2016 | DOI: 10.1002/bit.26013

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      Quartz crystal microbalance measurements suggest that a transcriptional regulator SpnR in Serratia marcescens AS-1 specifically interacts with guanine at position 5 on lux-box-like promoter spn box. A negative regulation of SpnR is involved with cell density dependent quorum sensing whose activation depends on the dissociation of SpnR from its binding site on DNA. The resonance frequency change shows that the SpnR binding affinity was reduced in the order of C16G and G5C substitution of the spn box.

  3. Communication to the Editor

    1. Improved production of N-acetylglucosamine in Saccharomyces cerevisiae by reducing glycolytic flux

      Sang-Woo Lee and Min-Kyu Oh

      Version of Record online: 3 JUN 2016 | DOI: 10.1002/bit.26014

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      Reducing glycolytic flux to improve GlcNAc production by disrupting PFK-2 in Saccharomyces cerevisiae was examined. Eliminating PFK-2 alone, which produces F26BP, allosteric activator of PFK-1, did not significantly change the GlcNAc production. When galactose was used for PFK-2 deleted strain, GlcNAc production was doubled, suggesting that significant reduction of glycolytic flux was needed for improving GlcNAc production.

  4. Articles

    1. Identifying the differences in mechanisms of mycophenolic acid controlling fucose content of glycoproteins expressed in different CHO cell lines

      An Zhang, Valerie Liu Tsang, Lam R. Markely, Lutfiye Kurt, Yao-Ming Huang, Shashi Prajapati and Rashmi Kshirsagar

      Version of Record online: 11 MAY 2016 | DOI: 10.1002/bit.25995

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      Understanding the fucosylation pathway and being able to control this process at the desired level are critical for bioproduction of highly efficient therapeutic glycoproteins. In this study, the authors explored several strategies and mycophenolic acid (MPA) was able to finely modulate the fucose content with the least undesired side effects. Significant differences in fucosylation levels of three tested recombinant CHO cell lines (CHO-DG44, CHO-DXB11, and CHO-S) were attributed to changes in the intracellular nucleotide involved in fucosylation pathway after MPA treatment due to differences in their MPA metabolism.

    2. Enhanced secretion of recombinant proteins via signal recognition particle (SRP)-dependent secretion pathway by deletion of rrsE in Escherichia coli

      Yong Jae Lee, Roojin Lee, Se Hwa Lee, Sung Sun Yim and Ki Jun Jeong

      Version of Record online: 11 MAY 2016 | DOI: 10.1002/bit.25997

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      The authors developed novel E. coli mutant of which secretion capacity of SRP pathway was fairly improved. Using this mutant, several useful therapeutic proteins including scFv, IgG, and GPCR, were successfully produced with high production yield.

    3. Chemical and morphological gradient scaffolds to mimic hierarchically complex tissues: From theoretical modeling to their fabrication

      Alessandra Marrella, Maurizio Aiello, Rodolfo Quarto and Silvia Scaglione

      Version of Record online: 8 MAY 2016 | DOI: 10.1002/bit.25994

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      A theoretical model to design and fabricate functional and structural complex graded scaffolds with predicted morphological and chemical features has been developed. The efficacy of the model has been tested by realizing engineered graded grafts for osteochondral tissue engineering applications. The procedure has been applied on both polycaprolactone (PCL) and collagen-type-I (COL) and a functional gradient was combined to the morphological one by adding hydroxyapatite (HA) powders, to mimic the bone mineral phase.

    4. Design and demonstration of a pumpless 14 compartment microphysiological system

      Paula G. Miller and Michael L. Shuler

      Version of Record online: 29 APR 2016 | DOI: 10.1002/bit.25989

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      The design and operation of a bidirectional flow, pumpless 14 chamber system representing 13 tissues/organs with both barrier and nonbarrier tissue chambers is described. Viability across all compartments was initially demonstrated with one cell line. Then viability and functionality in the device was retained for 7 days using five representative cell lines and compared to static controls.

    5. You have full text access to this OnlineOpen article
      Comprehensive genome and epigenome characterization of CHO cells in response to evolutionary pressures and over time

      Julia Feichtinger, Inmaculada Hernández, Christoph Fischer, Michael Hanscho, Norbert Auer, Matthias Hackl, Vaibhav Jadhav, Martina Baumann, Peter M. Krempl, Christian Schmidl, Matthias Farlik, Michael Schuster, Angelika Merkel, Andreas Sommer, Simon Heath, Daniel Rico, Christoph Bock, Gerhard G. Thallinger and Nicole Borth

      Version of Record online: 29 APR 2016 | DOI: 10.1002/bit.25990

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      Six closely related CHO cell lines were studied for their genome and epigenome modifications in response to evolutionary pressures and over time. Full genome and DNA methylation data including analyses for SNPs, InDels, translocations and other structural variants, full genome DNA methylation patterns, and detailed data on histone modifications during a batch culture are presented in a web database at cho-epigenome.boku.ac.at/.

    6. Highly selective production of succinic acid by metabolically engineered Mannheimia succiniciproducens and its efficient purification

      Sol Choi, Hyohak Song, Sung Won Lim, Tae Yong Kim, Jung Ho Ahn, Jeong Wook Lee, Moon-Hee Lee and Sang Yup Lee

      Version of Record online: 29 APR 2016 | DOI: 10.1002/bit.25988

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      Mannheimia succiniciproducens PALK strain (ΔldhA, Δpta-ackA) was constructed based on constraint-based flux analysis to produce nearly homo-succinic acid with a productivity high enough for economic industrial bio-based succinic acid production.

    7. Quantitative analysis of the supernatant from host and transfected CHO cells using iTRAQ 8-plex technique

      Guijie Zhu, Liangliang Sun, Thomas Albanetti, Travis Linkous, Christopher Larkin, Ronald Schoner, James B. McGivney IV and Norman J. Dovichi

      Version of Record online: 28 APR 2016 | DOI: 10.1002/bit.25991

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      Experimental design for 8-plex iTRAQ quantitative proteomic analysis of the expression difference of supernatant proteins from transfected clones and untransfected host cells. Experiments employed the CHO K1SV and CHO CAT-S cell lines. Three clones and the parent cell line were analyzed for both cell lines.

    8. Optimizing cell seeding and retention in a three-dimensional bioengineered cardiac ventricle: The two-stage cellularization model

      Nikita M. Patel, Iman K. Yazdi, Ennio Tasciotti and Ravi K. Birla

      Version of Record online: 28 APR 2016 | DOI: 10.1002/bit.25992

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      Cell seeding is pivotal in tissue engineering. This study develops the novel method of two-stage cellularization, combining direct cell injection of cardiac cells and the attachment of a fibrin gel artificial heart muscle patch, on a 3D bioengineered left ventricle scaffold. This strategy shows an increase in cell retention and biopotential function in the scaffold, compared to singular methods of cellularization. The presented concept is novel, with the potential to have a significant impact on the field of tissue engineering.

    9. Evolutionary engineering of Geobacillus thermoglucosidasius for improved ethanol production

      Jiewen Zhou, Kang Wu and Christopher V. Rao

      Version of Record online: 22 APR 2016 | DOI: 10.1002/bit.25983

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      The ability to grow at high temperatures makes thermophiles attractive for many fermentation processes. Here, the authors engineered a thermophilic bacterium, Geobacillus thermoglucosidasius, to more efficiently produce ethanol from glucose or cellobiose by using a combination of metabolic engineering and evolutionary engineering.

    10. Cell-free production and streamlined assay of cytosol-penetrating antibodies

      Seung Eui Min, Kyung-Ho Lee, Seong-Wook Park, Tae Hyeon Yoo, Chan Hee Oh, Ji-Ho Park, Sung Yun Yang, Yong-Sung Kim and Dong-Myung Kim

      Version of Record online: 21 APR 2016 | DOI: 10.1002/bit.25985

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      Antibodies that target intracellular proteins hold great promise in the development of novel therapeutic interventions for various diseases. In this study, cell-free protein synthesis technology was successfully applied for the generation of a cytosol-penetrating scFv. Compared to Escherichia coli production, a smaller reaction volume, minimum laboratory setup, and high-speed generation of more functional antibodies justify the use of cell-free protein synthesis as an advantageous method for developing cell-penetrating antibodies.

    11. Calorespirometric feeding control enhances bioproduction from toxic feedstocks—Demonstration for biopolymer production out of methanol

      Maria-Teresa Rohde, Sven Paufler, Hauke Harms and Thomas Maskow

      Version of Record online: 18 APR 2016 | DOI: 10.1002/bit.25986

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      A calorespirometric process control strategy for the conversion of toxic substrates into industry-relevant chemicals was developed and demonstrated exemplarily for growth of the methylotroph Methylobacterium extorquens on methanol. Feeding control is based on biocalorimetry for an online estimation of growth parameters such as the specific growth rate and uses calorespirometric data to distinguish between different microbial growth phases. Applying this approach, cultures reached considerably higher growth rates and product yields on methanol compared to different control strategies.

    12. Time-dependent combinatory effects of active mechanical loading and passive topographical cues on cell orientation

      Qian Wang, Hanyang Huang, Kang Wei and Yi Zhao

      Version of Record online: 14 APR 2016 | DOI: 10.1002/bit.25981

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      The combinatory effect of active (cyclic strain) and passive (linear surface wrinkles) mechanical stimulations on cell orientation was investigated. The results demonstrate that cyclic strain can reorient cells regardless of wrinkle size, while the reorienting behavior varies with strain magnitude and wrinkle size. Surface wrinkles can reorient the cells after strain removal, while the reorientation capability likewise varies with wrinkle size. The observation time point is thus critical for accurate interpretation of cell behavior under the combinatory mechanical stimulations.

    13. Plasmonic activation of gold nanorods for remote stimulation of calcium signaling and protein expression in HEK 293T cells

      Sandra P. Sanchez-Rodriguez, Jeremy P. Sauer, Sarah A. Stanley, Xi Qian, Andrew Gottesdiener, Jeffrey M. Friedman and Jonathan S. Dordick

      Version of Record online: 8 APR 2016 | DOI: 10.1002/bit.25984

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      The external and remote control of cellular behavior provides a powerful tool to manipulate biological process. Sanchez-Rodriguez et al. demonstrated that NIR-induced gold nanorod heating, through the selective targeting of membrane proteins (TRPV1 and integrins), can induce selective and temporal internalization of calcium in HEK 293T cells and controlled protein expression without causing cell damage.

    14. GroE chaperonins assisted functional expression of bacterial enzymes in Saccharomyces cerevisiae

      Peng-Fei Xia, Guo-Chang Zhang, Jing-Jing Liu, Suryang Kwak, Ching-Sung Tsai, In Iok Kong, Bong Hyun Sung, Jung-Hoon Sohn, Shu-Guang Wang and Yong-Su Jin

      Version of Record online: 6 APR 2016 | DOI: 10.1002/bit.25980

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      This study demonstrated that the mismatching of HSP60 chaperonins between bacteria and yeast is the reason that some bacterial enzymes cannot be functionally expressed in Saccharomyces cerevisiae. A post-translational tool is further developed to facilitating the functional expression of bacterial enzymes in S. cerevisiae via co-expression of bacterial groE chaperonins.

    15. Carboxypeptidase D is the only enzyme responsible for antibody C-terminal lysine cleavage in Chinese hamster ovary (CHO) cells

      Zhilan Hu, Henry Zhang, Benjamin Haley, Frank Macchi, Feng Yang, Shahram Misaghi, Joseph Elich, Renee Yang, Yun Tang, John C. Joly, Bradley R. Snedecor and Amy Shen

      Version of Record online: 6 APR 2016 | DOI: 10.1002/bit.25977

    16. Optimization of biomolecule separation by combining microscale filtration and design-of-experiment methods

      Amir S. Kazemi, Karina Kawka and David R. Latulippe

      Version of Record online: 31 MAR 2016 | DOI: 10.1002/bit.25975

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      In this work, a microscale filtration technique known as “stirred-well filtration” was combined with design-of-experiments (DOE) methods to enable the rapid determination of the effect of filtration process variables (e.g., permeate flux and solution pH) on the separation of biomolecules. The usefulness of this approach was demonstrated for three model mixtures of biomolecules. The filtration performance metrics are presented in a unique format to match the factorial design of the DOE method.

    17. Co-production of acetone and ethanol with molar ratio control enables production of improved gasoline or jet fuel blends

      Zachary C. Baer, Sebastian Bormann, Sanil Sreekumar, Adam Grippo, F. Dean Toste, Harvey W. Blanch and Douglas S. Clark

      Version of Record online: 31 MAR 2016 | DOI: 10.1002/bit.25978

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      Escherichia coli was metabolically engineered for the co-production of acetone and ethanol. Coupling of these fermentation products via catalytic alkylation affords a low-toxicity route for the production of advanced bio-gasoline and jet fuel blendstocks of carbon length C5–C15. The authors demonstrate how to alter the ethanol:acetone molar ratio by engineering the metabolic pathway and environmental redox state to control the end-product distribution after chemical catalysis.

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