Biotechnology and Bioengineering

Cover image for Vol. 113 Issue 11

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. Discriminating typical and atypical cystic fibrosis-related bacteria by multiplex PNA-FISH

      Susana P. Lopes, Daniel T. Carvalho, Maria O. Pereira and Nuno F. Azevedo

      Version of Record online: 26 SEP 2016 | DOI: 10.1002/bit.26085

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      This study highlights the successful adaptation of a multiplex PNA-FISH assay to detect polymicrobial communities encompassing CF-typical and atypical bacteria. The method is therefore suitable to further describe species-dependent prevention strategies and deliver more effective target control therapeutics in cystic fibrosis disease.

  2. Reviews

    1. The future of protein scaffolds as affinity reagents for purification

      Ana M.G.C. Dias and Ana C.A. Roque

      Version of Record online: 26 SEP 2016 | DOI: 10.1002/bit.26090

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      In this review we highlight the developments of small protein scaffolds as reagents in affinity purification. We discuss the biophysical properties, protein evolution, and engineering strategies of non-immunoglobulin scaffolds; as well as, their targets, immobilization methodologies, solid supports, and purification conditions. We emphasize the strengths and weaknesses of the scaffolds versus current affinity reagents. Finally, we explore the challenges and future opportunities for these scaffolds to be implemented as affinity ligands in research and industrial applications.

  3. Communications to the Editor

    1. Fam60A plays a role for production stabilities of recombinant CHO cell lines

      Anett Ritter, Sandro Nuciforo, Axel Schulze, Mevion Oertli, Tatjana Rauschert, Bernd Voedisch, Sabine Geisse, Thomas Jostock and Holger Laux

      Version of Record online: 22 SEP 2016 | DOI: 10.1002/bit.26181

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      Ritter and coauthors provide primary evidence that the FAM60A protein plays a key role in production stability of CHO cell lines. Knock-out of the gene Fam60A, which is located within the telomeric region of the small chromosome 8 in CHO-K1a cells, increases the percentage of stable clones after long-term cultivation. This highlights that Fam60a is a relevant gene for cell line engineering approaches to increase production stability in recombinant cell lines.

  4. Articles

    1. Production of citramalate by metabolically engineered Escherichia coli

      Xianghao Wu and Mark A. Eiteman

      Version of Record online: 22 SEP 2016 | DOI: 10.1002/bit.26035

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      Escherichia coli with knockouts in gltA, ackA, and leuC genes expressing citramalate synthase was used to generate the methacrylic acid precursor, citramalate. A fed-batch process using a glucose feed resulted in 46.5 g/L citramalate in 132 h with a yield of 0.63 g/g, over 75% of theoretical maximum.

    2. Non-chromatographic bioprocess engineering of a recombinant mineralizing protein for the synthesis of silica nanocapsules

      David Wibowo, Guang-Ze Yang, Anton P.J. Middelberg and Chun-Xia Zhao

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26079

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      A chromatography-free and high-yield recovery of mineralizing protein, D4S2, from recombinant Escherichia coli for making silica nanocapsules was developed based on selective thermochemical precipitation method: (i) DNA removal using poly(ethyleneimine); (ii) protein-contaminant precipitation using 1 M Na2SO4 (90°C, 5 min) in which D4S2 protein remained soluble; and (iii) D4S2-protein isolation using 1.8 M Na2SO4 (30°C, 60 min). The final yield of D4S2 protein was ∼29 mg per 800 mL cells (OD600 ∼2) which accounts for ∼79% of D4S2-protein recovery.

  5. Reviews

    1. Stem cells for spinal cord injury: Strategies to inform differentiation and transplantation

      Nisha R. Iyer, Thomas S. Wilems and Shelly E. Sakiyama-Elbert

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26074

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      Advances in stem cell technologies have enabled their ethical and efficacious use in the context of spinal cord injury treatment. This review describes strategies to optimize the differentiation of stem cells into discrete spinal cell types, including culture formats, induction methodologies, and the directed differentiation of somatic cells. In addition to outcomes post-transplantation, the authors discuss potential combinatorial strategies with biomaterial scaffolds and controlled drug delivery systems to improve stem cell therapies for clinical use.

  6. Articles

    1. You have full text access to this OnlineOpen article
      Real-time monitoring and control of the load phase of a protein A capture step

      Matthias Rüdt, Nina Brestrich, Laura Rolinger and Jürgen Hubbuch

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26078

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      Protein A capture steps are central to the preparative purification of mAbs. However, capture steps are only partly controlled in realtime. In this study, partial least squares regression on UV/Vis absorption spectra was applied to quantify mAb in the effluent of a protein A capture step during the load phase.

    2. Comparative thermal inactivation analysis of Aspergillus oryzae and Thiellavia terrestris cutinase: Role of glycosylation

      Abhijit N. Shirke, An Su, J. Andrew Jones, Glenn L. Butterfoss, Mattheos A.G. Koffas, Jin Ryoun Kim and Richard A. Gross

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26052

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      An in-depth understanding of the thermal inactivation mechanism of proteins provides guidelines for their thermostabilization. Here, Shirke and coworkers proposed a unified model for the thermal inactivation of Cutinases (Polyester hydrolases) based on comparative analysis of homologues. Considering a prominent role aggregation in thermal inactivation of these enzymes, the authors have proposed glycosylation site engineering as a strategy for the inhibition of thermal aggregation which was found to be better than conventional use of trehalose under a typical condition.

    3. Optimization of an acetate reduction pathway for producing cellulosic ethanol by engineered yeast

      Guo-Chang Zhang, In Iok Kong, Na Wei, Dairong Peng, Timothy L. Turner, Bong Hyun Sung, Jung-Hoon Sohn and Yong-Su Jin

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26021

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      Acetate, a major fermentation inhibitor present in lignocellulosic hydrolysate, can be co-consumed with xylose synergistically to produce ethanol via an acetate reduction pathway, allowing both efficient xylose fermentation and in situ detoxification of acetate in lignocellulosic hydrolysate. Enzymatic activities of ACS and AADH were identified as limiting factors of the acetate reduction pathway, and optimization of the expression levels of ACS and AADH led to enhanced co-fermentation of xylose and acetate.

    4. Synthesis of bolaform biosurfactants by an engineered Starmerella bombicola yeast

      Inge N. A. Van Bogaert, Dieter Buyst, José C. Martins, Sophie L. K. W. Roelants and Wim K. Soetaert

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26032

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      Bolaform surfactants recently attract a lot of attention due to their unique configuration consisting of a long hydrophobic spacer connecting two instead of one hydrophilic entities. Currently, these molecules are produced by chemical means. We established an efficient microbial production process for the fully green synthesis of bolaform surfactants. The sophorolipid producing yeast Starmerella bombicola was disabled in its sophorolipid acetyltransferase and lactone esterase, resulting in synthesis of a mixture of classic amphiphilic structures and large amounts of novel bola-amphiphiles.

    5. Cellobiose dehydrogenase and chitosan-based lysozyme responsive materials for antimicrobial wound treatment

      Christoph Öhlknecht, Gregor Tegl, Bianca Beer, Christoph Sygmund, Roland Ludwig and Georg M. Guebitz

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26070

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      General scheme of the mode of action of the intelligent antimicrobial system. NAc-CTS is hydrolyzed to COS in the presence of lysozyme. The produced COSs act as substrate for CDH that generates the antimicrobial agent H2O2.

    6. Continuous multistep synthesis of perillic acid from limonene by catalytic biofilms under segmented flow

      Christian Willrodt, Babu Halan, Lisa Karthaus, Jessica Rehdorf, Mattijs K. Julsing, Katja Buehler and Andreas Schmid

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26071

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      Catalytic biofilms in segmented flow capillary micro-reactors enabled successful multi-step oxyfunctionalization of (R)-(+)-limonene to (R)-(+)-perillic acid with a high productivity of 34 g Ltube−1 d−1. We demonstrated the lab-scale production of a highly toxic chemical from a volatile substrate with a wild type Pseudomonas putida GS1 and as well a recombinant Pseudomonas taiwanensis VLB120 harboring p-cymene monooxygenase. The technical applicability of the concept was extended by purifying the product. The conceptual design can be transferred to other equally challenging biotransformations.

    7. Loop variants of the thermophile Rasamsonia emersonii Cel7A with improved activity against cellulose

      Trine Holst Sørensen, Michael Skovbo Windahl, Brett McBrayer, Jeppe Kari, Johan Pelck Olsen, Kim Borch and Peter Westh

      Version of Record online: 21 SEP 2016 | DOI: 10.1002/bit.26050

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      Engineering of one the loops covering the active site tunnel of Cellobiohydrolase I from Rasamsonia emersonii reveals an inverse affinity/activity relationship. The authors show that replacement of N194 and particularly N197 with alanine results in variants with significantly reduced substrate affinity and higher catalytic efficacy. By fitting a steady state model for processive enzyme to data, it is suggested that the mutations led to higher enzyme-substrate dissociation but did not affect the rate of association.

    8. A pumpless multi-organ-on-a-chip (MOC) combined with a pharmacokinetic–pharmacodynamic (PK–PD) model

      Hyuna Lee, Dae Shik Kim, Sang Keun Ha, Inwook Choi, Jong Min Lee and Jong Hwan Sung

      Version of Record online: 14 SEP 2016 | DOI: 10.1002/bit.26087

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      Multi-organ-on-a-chip recapitulates simultaneous metabolism and cell-killing effect of drugs, and its action inside the chip is simulated with a PK–PD model, providing insight into the action of the drug.

    9. LPMOs in cellulase mixtures affect fermentation strategies for lactic acid production from lignocellulosic biomass

      Gerdt Müller, Dayanand Chandrahas Kalyani and Svein Jarle Horn

      Version of Record online: 9 SEP 2016 | DOI: 10.1002/bit.26091

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      Bioconversion of birch wood to lactic acid by means of SHF and SSF using an LPMO-containing cellulase cocktail combined with either a mesophilic or thermophilic fermenting microbe.

  7. Communication to the Editors

    1. Application of an oxygen-inducible nar promoter system in metabolic engineering for production of biochemicals in Escherichia coli

      Hee J. Hwang, Jin W. Kim, Si Y. Ju, Jin H. Park and Pyung C. Lee

      Version of Record online: 7 SEP 2016 | DOI: 10.1002/bit.26082

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      A dissolved oxygen-dependent nar promoter has been utilized as an alternative to commonly-used promoters for engineering biochemical pathways because of its relatively simple and cost-effective induction mechanism. The three engineered biosynthetic pathways in which the expression of genes was controlled by the nar promoter showed reasonable performances [113.12 ± 2.37 g/L d-lactate, 48.0 ± 8.48 g/L (R, R)-2,3-butandiol, and 15.8 ± 0.62 g/L 1, 3-propandiol].

  8. Reviews

    1. Modulation of human mesenchymal and pluripotent stem cell behavior using biophysical and biochemical cues: A review

      Sheryl Ding, Peter Kingshott, Helmut Thissen, Martin Pera and Peng-Yuan Wang

      Version of Record online: 2 SEP 2016 | DOI: 10.1002/bit.26075

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      The ability to manipulate human stem cells in vitro is vital in regenerative medicine and cellular therapies. While biochemical cues have been the focus of this area for some time, biophysical cues are emerging as a major factor in stem cell control. In this review, the authors summarize how different forms of biophysical stimulation influence stem cell behavior, and compare their effects to those of conventional culture methods.

  9. Articles

    1. Mixing and mass transfer in a pilot scale U-loop bioreactor

      Leander A.H. Petersen, John Villadsen, Sten B. Jørgensen and Krist V. Gernaey

      Version of Record online: 2 SEP 2016 | DOI: 10.1002/bit.26084

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      Mixing time and mass transfer coefficients considerably higher than those obtainable in an STR and previous tubular loop reactors were determined in a forced flow U-loop fermenter of a novel construction. The system also proved capable of handling a large volumetric gas fraction, which was shown to have only minor effect on the power drawn from the pump impeller.

    2. Efficient gene knockdown in Clostridium acetobutylicum by synthetic small regulatory RNAs

      Changhee Cho and Sang Yup Lee

      Version of Record online: 30 AUG 2016 | DOI: 10.1002/bit.26077

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      Clostridium acetobutylicum is an industrially important bacterium, that has been known to be extremely difficult to genetically manipulate. Here we report development of synthetic sRNA-based gene knockdown system that allows efficient knockdown of inherent and heterologous genes in C. acetobutylicum. By knocking down the pta gene in buk-negative mutant C. acetobutylicum strain, butanol production could be enhanced both in batch culture and fed-batch culture coupled with in situ recovery.

    3. Boosted structured additive regression for Escherichia coli fed-batch fermentation modeling

      Michael Melcher, Theresa Scharl, Markus Luchner, Gerald Striedner and Friedrich Leisch

      Version of Record online: 30 AUG 2016 | DOI: 10.1002/bit.26073

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      In this study, the potential of boosted structured additive regression (STAR) models for monitoring key process parameters such as the cell dry mass or protein concentration in Escherichia coli fed-batch fermentations is investigated. Besides promising results—the responses can be predicted with an accuracy between 3% and 16%—boosted STAR models are more interpretable and facilitate the assessment of predictor importance.