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Biotechnology Journal

Zelun Zhi, Imad Younus Hasan, Adam Mechler
A phospholipid monolayer supported on an alkanethiol self-assembled monolayer (SAM) constitutes a supported hybrid membrane, a model of biological membranes optimized for electronic access through the underlying metal support surface. It is believed that phospholipids, when deposited from aqueous liposome suspension, spontaneously cover the alkanethiol-modified surface, owing to the reduction of surface free energy of the hydrophobic alkane surface exposed to the solution. However, the formation of the hybrid layer has to overcome significant energy barriers in rupturing the vesicle and "unzipping" the membrane leaflets; hence drivers of the spontaneous hybrid membrane formation are unclear...
July 14, 2018: Biotechnology Journal
Patrick C F Buchholz, Ruediger Ohs, Antje C Spiess, Juergen Pleiss
The estimation of kinetic parameters provides valuable insights into the function of biocatalysts and is indispensable in optimizing process conditions. Frequently, kinetic analysis relies on the Michaelis-Menten model derived from initial reaction rates at different initial substrate concentrations. However, by analysis of complete progress curves, more complex kinetic models can be identified. This case study compares two previously published experiments on benzaldehyde lyasecatalyzed self-ligation for the substrates benzaldehyde and 3,5-dimethoxybenzaldehyde to investigate (1) the effect of using different kinetic model equations on the kinetic parameter values, and (2) the effect of using models with and without enzyme inactivation on the kinetic parameter values...
July 12, 2018: Biotechnology Journal
Alan Costello, Nga T Lao, Clair Gallagher, Berta Capella-Roca, Lourdes A N Julius, Srinivas Suda, Jens Ducrée, Damien King, Roland Wagner, Niall Barron, Martin Clynes
With the ability to affect multiple genes and fundamental pathways simultaneously, miRNA engineering of Chinese Hamster Ovary (CHO) cells has significant advantages over single gene expression or repression. Tight control of these molecular triggers is desirable as it could in theory allow on/ off or even tunable regulation of desirable cellular phenotypes. The present study investigated the potential of employing a tetracycline inducible (TET-On) system for conditional knockdown of specific miRNAs but encountered several challenges...
July 10, 2018: Biotechnology Journal
Lyne Jossé, Lin Zhang, C Mark Smales
The DHFR system is used for the selection of recombinant CHO cell lines using the inhibitor MTX. During clonal selection, endogenous DHFR expression and resistance to MTX allows the selection of cells expressing sufficient DHFR to survive. Here we describe a novel vector platform for the DHFR system, whereby addition of a synthetic 3'UTR destabilizes DHFR expression. We harnessed miRs ability to negatively regulate gene expression by their near-complementary binding to the 3'UTR region of transcripts. From the literature we identified let-7f as a highly abundant, invariant miR in CHO cells...
July 7, 2018: Biotechnology Journal
Musashi Takenaka, Jae Min Lee, Prihardi Kahar, Chiaki Ogino, Akihiko Kondo
Actinobacteria plays a key role in the cycling of organic matter in soils. They secret biomass-degrading enzymes that allow it to produce the unique metabolites that originate in plant biomass. Although past studies have focused on these unique metabolites, a large-scale screening of Actinobacteria is yet to be reported to focus on their biomass-degrading ability. In the present study, a rapid and simple method was constructed for a large-scale screening, and the novel resources that form the plant biomass-degrading enzyme cocktail were identified from 850 isolates of Actinobacteria...
July 6, 2018: Biotechnology Journal
Jia Wang, Thomas Scheibel
Mussel byssus, the holdfast of mussels, has attracted much attention due to the mechanical gradient properties of its threads and the robust and flexible adhesion to various substrates, features that originating mainly from mussel byssus proteins and could be used in potential applications in biomedicine. However, the applications have been impeded owing to the difficulties concerning the extraction of the natural materials, i.e. proteins. Recent advances in recombinant production provide a novel approach to generate mussel byssus inspired proteins, even with the opportunities for scale-up production...
July 5, 2018: Biotechnology Journal
Chiara Lonoce, Carla Marusic, Elena Morrocchi, Anna Maria Salzano, Andrea Scaloni, Flavia Novelli, Claudio Pioli, Mistianne Feeney, Lorenzo Frigerio, Marcello Donini
Hairy root (HR) cultures represent an attractive platform for the production of heterologous proteins, due to the possibility of secreting the molecule of interest in the culture medium. The main limitation is the low accumulation yields of heterologous proteins. The aim of this study was to enhance the accumulation of a tumor-targeting antibody with a human-compatible glycosylation profile in HR culture medium. To this aim, we produced Nicotiana benthamiana HR cultures expressing the red fluorescent protein (RFP) to easily screen for different auxins able to induce heterologous protein secretion in the medium...
July 5, 2018: Biotechnology Journal
Leila Zamani, Magnus Lundqvist, Ye Zhang, Magnus Aberg, Fredrik Edfors, Gholamreza Bidkhori, Anna Lindahl, Axel Mie, Adil Mardinoglu, Raymond Field, Richard Turner, Johan Rockberg, Veronique Chotteau
The optimization of bioprocesses for biopharmaceutical manufacturing by Chinese hamster ovary (CHO) cells can be a challenging endeavor and, today, heavily relies on empirical methods treating the bioreactor process and the cells as black boxes. Multi-omics approaches have the potential to reveal otherwise unknown characteristics of these systems and identify culture parameters to more rationally optimize the cultivation process. Here we have applied both metabolomic and proteomic profiling to a perfusion process, using CHO cells for antibody production, to explore how cell biology and reactor environment change as the cell density reaches ≥ 200 x 106 cells/mL...
June 29, 2018: Biotechnology Journal
Ningning Yang, Wenwen Zhang, Chao Ye, Xue Chen, Shengjie Ling
Living organisms produced nanopolymers (nanobiopolymers for short), such as nanocellulose, nanochitin, nanosilk, nanobiotch, and microbial nanobiopolymers, having received widely scientific and engineering interests in recent years. Compare with petroleum-based polymers, biopolymers are sustainable and biodegradable. The unique structural features that stem from nanosized effects, such as ultrahigh aspect ratio and length-diameter ratio, further endow nanobiopolymers with high transparence and versatile processability...
June 28, 2018: Biotechnology Journal
Fang Yang, Ying Zhang, Xi Cui, Yutong Fan, Ying Xue, Haipeng Miao, Guiying Li
Blood is the most valuable and convenient source of disease diagnostic biomarkers, but plasma must be extracted from blood prior to further biomarker detection and analysis. Traditional plasma extraction methods depend greatly on benchtop equipment and are often cumbersome, time-consuming, and unsuitable for point-of-care (POC) applications. On the other hand, highly diluted blood plasma extraction methods potentially introduce greater test result uncertainty and lower detection sensitivity. Because simpler integrated and more efficient plasma extraction methods that avoid sample dilution are highly desired, we developed a simple dielectrophoresis (DEP)-based microfluidic device for plasma extraction from whole blood...
June 28, 2018: Biotechnology Journal
Christian Lieven, Markus J Herrgård, Nikolaus Sonnenschein
Developing methylotrophic bacteria into cell factories that meet the chemical demand of the future could be both economical and environmentally friendly. Methane is not only an abundant, low-cost resource but also a potent greenhouse gas, the capture of which could help to reduce greenhouse gas emissions. Rational strain design workflows rely on the availability of carefully combined knowledge often in the form of genome-scale metabolic models to construct high-producer organisms. In this review, we present the most recent genome-scale metabolic models in aerobic methylotrophy and their applications...
June 19, 2018: Biotechnology Journal
Аnatoly A Abalymov, Roman A Verhovskiy, Marina V Novoselova, Bogdan V Parakhonskiy, Dmitry A Gorin, Alexey M Yashchenok, Gleb B Sukhorukov
Porous calcium carbonate (CaCO3 ) vaterite particles are very attractive templates for the encapsulation of pharmaceuticals and for the construction of hollow polyelectrolyte capsules, sensors, and enzyme-catalyzed reactors. Although CaCO3 is biocompatible and biodegradable, little is known about the intercellular behavior and properties of vaterite particles in the cytoplasm of cells. In this work, we combined confocal Raman and fluorescent microscopy for the imaging of porous CaCO3 vaterite particles in HeLa cells to study the uptake and status of the particles inside the cells in real time...
June 19, 2018: Biotechnology Journal
Mingfeng Cao, Meirong Gao, Deon Ploessl, Cunjiang Song, Zengyi Shao
Scheffersomyces stipitis, renowned for its native xylose-utilizing capacity, has recently demonstrated its potential in producing health-promoting shikimate pathway derivatives. However, its broader application is hampered by the low transformation efficiency and the lack of genetic engineering tools to enable sophisticated genomic manipulations. S. stipitis employees the predominant non-homologous end joining (NHEJ) mechanism for repairing DNA double-strand breaks (DSB), which is less desired due to its incompetence in achieving precise genome editing...
June 19, 2018: Biotechnology Journal
Katia Tarasava, Eun Joong Oh, Carrie A Eckert, Ryan T Gill
In recent years CRISPR-Cas technologies have revolutionized microbial engineering approaches. Genome editing and non-editing applications of various CRISPR-Cas systems have expanded the throughput and scale of engineering efforts, as well as opened up new avenues for manipulating genomes of non-model organisms. As we expand the range of organisms used for biotechnological applications, we need to develop better, more versatile tools for manipulation of these systems. Here we summarize the current advances in microbial gene editing using CRISPR-Cas based tools, and highlight state-of-the-art methods for high-throughput, efficient genome-scale engineering in model organisms Escherichia coli and Saccharomyces cerevisiae...
June 19, 2018: Biotechnology Journal
Linas Tamošaitis, C Mark Smales
Transcriptomics has been extensively applied to the investigation of the CHO cell platform for the production of recombinant biotherapeutic proteins to identify transcripts whose expression is regulated and correlated to (non)desirable CHO cell attributes. However, there have been few attempts to analyse the findings across these studies to identify conserved changes and generic targets for CHO cell platform engineering. Here we have undertaken a meta-analysis of CHO cell transcriptomic data and report on those genes most frequently identified as differentially expressed with regard to cell growth (µ) and productivity (Qp)...
June 19, 2018: Biotechnology Journal
Hui Tao, Yuchen Zhang, Zixin Deng, Tiangang Liu
Spinosad is a potent insecticide that exhibits an excellent environmental and mammalian profile. However, spinosad production in the original producer, Saccharopolyspora spinosa, is insufficient for the huge global demand. Great efforts have been exerted to improve the production of spinosad. Strategies for spinosad overproduction in actinomycetes were reviewed in this article, including metabolic engineering of the precursor and spinosyn biosynthetic pathway, introduction of regulatory genes, genome-scale metabolic model-guided engineering, mutagenesis, genome shuffling, fermentation process optimization, omics analysis, and the heterologous biosynthesis of spinosad in other actinomycetes...
June 13, 2018: Biotechnology Journal
Greta Nölke, Mirna Barsoum, Marcel Houdelet, Elsa Arcalís, Fritz Kreuzaler, Rainer Fischer, Stefan Schillberg
Increasing the productivity of crops is a major challenge in agricultural research. Given that photosynthetic carbon assimilation is necessary for plant growth, enhancing the efficiency of photosynthesis is one strategy to boost agricultural productivity. We attempted to increase the photosynthetic efficiency and biomass of tobacco plants by expressing individual components of the Chlamydomonas reinhardtii carbon concentration mechanism (CCM) and integrating them into the chloroplast. Independent transgenic varieties were generated accumulating the carbonic anhydrase CAH3 in the thylakoid lumen or the bicarbonate transporter LCIA in the inner chloroplast membrane...
June 11, 2018: Biotechnology Journal
Mario A Torres-Acosta, Karla Mayolo-Deloisa, José González-Valdez, Marco Rito-Palomares
Aqueous two-phase systems (ATPS) have proved to be an efficient and integrative operation to enhance recovery of industrially-relevant bioproducts. After ATPS discovery, a variety of works have been published regarding their scaling from 10 to 1,000 liters. Although ATPS have achieved high recovery and purity yields, there is still a gap between their bench-scale use and potential industrial applications. In this context, this review paper, critically analyzed ATPS scale-up strategies to enhance the potential industrial adoption...
June 7, 2018: Biotechnology Journal
Liqiong Liu, Yizhou Zhang, Dongxu Jiang, Shiming Du, Zixin Deng, Lianrong Wang, Shi Chen
Bacterial epigenetic modifications play key roles in cellular processes such as stress responses, DNA replication, segregation, antimicrobial resistance, etc. In recent years, emerging new sequencing technologies, including single-molecule real-time (SMRT) sequencing and nanopore sequencing, have enabled the directed reading of epigenetic modifications without pre-treatment of DNA or DNA amplification. The applications of SMRT and nanopore sequencing open the door for the identification of more diverse epigenetic modifications of DNA bases and backbones and potentially facilitate the understanding of the novel functions of these epigenetic markers in cell physiology...
June 7, 2018: Biotechnology Journal
Donatella Lucchetti, Andrea Fattorossi, Alessandro Sgambato
Background The possibility to study solid tumors through the analysis of extracellular vesicles in biological fluids is one of the most exciting and rapidly advancing field in cancer research. The extracellular vesicles are tiny sacs released in both physiological and pathological conditions and could be used to monitor the evolution of several pathological states, including neoplastic diseases. Indeed, these vesicles carry biological informations and can affect the behavior of recipient cells by transferring proteins, DNA, RNA and microRNA...
June 7, 2018: Biotechnology Journal
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