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Biotechnology Journal

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https://www.readbyqxmd.com/read/28734022/3d-printed-vascularized-device-for-subcutaneous-transplantation-of-human-islets
#1
Marco Farina, Andrea Ballerini, Daniel Fraga, Eugenia Nicolov Matthew Hogan, Danilo Demarchi, Francesco Scaglione, Omaima Sabek, Philip Horner, Usha Thekkedath, Osama Gaber, Alessandro Grattoni
Transplantation of pancreatic islets or stem cell derived insulin secreting cells is an attractive treatment strategy for diabetes. However, islet transplantation is associated with several challenges including function-loss associated with dispersion and limited vascularization as well as the need for continuous immunosuppression. To overcome these limitations, here we present a novel 3D printed and functionalized encapsulation system for subcutaneous engraftment of islets or islet like cells. The devices were 3D printed with polylactic acid and the surfaces treated and patterned to increase the hydrophilicity, cell attachment and proliferation...
July 22, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731643/transcriptome-based-identification-of-the-optimal-reference-cho-genes-for-normalisation-of-qpcr-data
#2
Adam J Brown, Suzanne Gibson, Diane Hatton, David C James
Real-time quantitative PCR (qPCR) is the standard method for determination of relative changes in mRNA transcript abundance. Analytical accuracy, precision, and reliability are critically dependent on the selection of internal control reference genes. In this study we have identified optimal reference genes that can be utilized universally for qPCR analysis of CHO cell mRNAs. Initially, transcriptomic datasets were analysed to identify eight endogenous genes that exhibited high expression stability across four distinct CHO cell lines sampled in different culture phases...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731632/macrophages-of-different-phenotypes-influence-the-migration-of-bmscs-in-plga-scaffolds-with-different-pore-size
#3
Yuankun Dai, Xuguang Li, Ruihan Wu, Ying Jin, Changyou Gao
Immune modulation of the scaffolds not only reduces the host immunological rejection response, but also improves the regenerative cell migration into the scaffolds. Herein a convenient immune modulation of poly(lactide-co-glycolide) (PLGA) scaffold was applied with macrophages of different phenotypes to evaluate its influence on the migration behavior of bone marrow mesenchymal stem cells (BMSCs). With pro-inflammatory macrophages (M1) pre-loading, BMSCs migrated significantly faster into the PLGA scaffold, compared with those in the control scaffold or pre-seeded with inactivated macrophages (M0)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731631/substrate-curvature-restricts-spreading-and-induces-differentiation-of-human-mesenchymal-stem-cells
#4
Sang Joo Lee, Shengyuan Yang
While cells attach, spread, migrate, proliferate, and differentiate in three-dimensional (3D) micromechanical environments, the mechanical factors of these environments influence the shapes, sizes, and adhesion forces of the cells. Here, we culture human mesenchymal stem cells (hMSCs) on a unique class of curvature-defined substrates, micro glass ball embedded polyacrylamide gels, prepared with an improved protocol, and we investigate the spreading responses of the hMSCs on the glass balls to study the effects of substrate curvature on the spreading of hMSCs...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731630/improved-pks-gene-expression-with-strong-endogenous-promoter-resulted-in-geldanamycin-yield-increase
#5
Xinran Wang, Xinjuan Ning, Qinqin Zhao, Qianjin Kang, Linquan Bai
The type I polyketide geldanamycin is a potent anti-tumor reagent. Its biosynthesis includes three steps: the biosynthesis of precursors, such as 3-amino-5-hydroxybenzoic acid (AHBA), the PKS chain extension, and the post-PKS modifications. According to the genomic and transcriptomic analysis, the PKS chain extension was deduced to be the rate-limiting step for geldanamycin production in Streptomyces hygroscopicus XM201. in order to improve the expression of PKS genes, a strong endogenous promoter 5063p was obtained based on the transcriptomic analysis and XylE enzymatic assay...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731627/his-tag-immobilization-of-cutinase-1-from-thermobifida-cellulosilytica-for-solvent-free-synthesis-of-polyesters
#6
Alessandro Pellis, Marco Vastano, Felice Quartinello, Enrique Herrero Acero, Georg M Guebitz
For many years, lipase B from Candida antarctica (CaLB) was the primary biocatalyst used for enzymatic esterification and polycondensation reactions. More recently, the need for novel biocatalysts with different selectivity has arisen in the biotechnology and biocatalysis fields. The present work describes how the catalytic potential of Thermobifida cellulosilytica cutinase 1 (Thc_Cut1) was exploited for polyester synthesis. in a first step, Thc_Cut1 was immobilized on three different carriers, namely Opal, Coral and Amber, using a novel non-toxic His-tag method based on chelated Fe(iii) ions (>99% protein bounded)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731622/mechanistic-modeling-based-pat-implementation-for-ion-exchange-process-chromatography-of-charge-variants-of-monoclonal-antibody-products
#7
Vijesh Kumar, Anurag S Rathore
Process chromatography is typically used to remove product related impurities and variants that have very similar physicochemical properties to the product. Baseline separation may not be achieved in most cases due to high protein loading and thus, pooling of the elution peak can be challenging for maximizing yield and achieving consistency in product quality. Batch-to-batch variability in quality of the feed material also occurs in commercial manufacturing. Mechanistic modeling of process chromatography, though non-trivial, can be an enabler for implementation of Process Analytical Technology...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731621/metabolic-engineering-for-advanced-biofuels-production-and-recent-advances-towards-commercialization
#8
Corey W Meadows, Aram Kang, Taek Soon Lee
Research on renewable biofuels produced by microorganisms has enjoyed considerable advances in academic and industrial settings. As the renewable ethanol market approaches maturity, the demand is rising for the commercialization of more energy-dense fuel targets. Many strategies implemented in recent years have considerably increased the diversity and number of fuel targets that can be produced by microorganisms. Moreover, strain optimization for some of these fuel targets has ultimately led to their production at industrial scale...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731613/polyol-structure-influences-enzymatic-hydrolysis-of-bio-based-2-5-furandicarboxylic-acid-fdca-polyesters
#9
Karolina Haernvall, Sabine Zitzenbacher, Hassan Amer, Michael Thomas Zumstein, Michael Sander, Kristopher McNeill, Motonori Yamamoto, Michael Bernhard Schick, Doris Ribitsch, Georg M Guebitz
Polyesters of 2,5-furan dicarboxylic acid (FDCA) have gained attention since they can be regarded as the bio-based alternatives to the petroleum based polyesters of terephthalic acid. However, only little is known about the biodegradation and enzymatic hydrolysis of FDCA-based polyesters. This work aimed to investigate the influence of different polyols on enzymatic hydrolysis of FDCA-based polyesters. A series of polyesters containing various polyols were synthetized and analysed regarding susceptibility to enzymatic hydrolysis by cutinase 1 from Thermobifida cellulosilytica (Thc_Cut1)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731612/mesoderm-lineage-3d-tissue-constructs-are-produced-at-large-scale-in-a-3d-stem-cell-bioprocess
#10
Jae Min Cha, Athanasios Mantalaris, Sunyoung Jung, Yurim Ji, Oh Young Bang, Hojae Bae
Various studies have presented different approaches to direct pluripotent stem cell differentiation such as applying defined sets of exogenous biochemical signals and genetic/epigenetic modifications. Although differentiation to target lineages was successfully regulated, such conventional methods are often complicated, laborious, and not cost-effective to be employed to the large-scale production of 3D stem cell-based tissue constructs. In this study, we developed a 3D-culture platform that could realize the large-scale production of mesoderm lineage tissue constructs from embryonic stem cells (ESCs)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731580/rewiring-the-glucose-transportation-and-central-metabolic-pathways-for-overproduction-of-n-acetylglucosamine-in-bacillus-subtilis
#11
Yang Gu, Jieying Deng, Yanfeng Liu, Jianghua Li, Hyun-Dong Shin, Guocheng Du, Jian Chen, Long Liu
N-acetylglucosamine (GlcNAc) is an important amino sugar extensively used in the healthcare field. in a previous study, we constructed the recombinant Bacillus subtilis strain BSGN6-PxylA -glmS-pP43NMK-GNA1 (BN0-GNA1) for microbial production of GlcNAc by pathway design and modular optimization. Here, we further improved the production of GlcNAc by rewiring both the glucose transportation and central metabolic pathways. First, the phosphotransferase system (PTS) was blocked by deletion of three genes, yyzE (encoding the PTS system transporter subunit iiA YyzE), ypqE (encoding the PTS system transporter subunit iiA YpqE), and ptsG (encoding the PTS system glucose-specific EiiCBA component), resulting in 47...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731574/rapid-buffer-and-ligand-screening-for-affinity-chromatography-by-multiplexed-surface-plasmon-resonance-imaging
#12
Karin P M Geuijen, Daniëlle E J W van Wijk-Basten, David F Egging, Richard B M Schasfoort, Michel H Eppink
Protein purifications are often based on the principle of affinity chromatography, where the protein of interest selectively binds to an immobilized ligand. The development of affinity purification requires selecting proper wash and elution conditions. In recent years, miniaturization of the purification process is applied to speed up the development (e.g. microtiterplates, robocolumns). We have studied the application of surface plasmon resonance imaging (SPRi) as a tool to simultaneously screen many buffer conditions for wash and elution steps in an affinity-based purification process...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731571/model-based-investigation-on-the-mass-transfer-and-adsorption-mechanisms-of-mono-pegylated-lysozyme-in-ion-exchange-chromatography
#13
Josefine Morgenstern, Gang Wang, Pascal Baumann, Jürgen Hubbuch
Recent studies highlighted the potential of PEGylated proteins to improve stabilities and pharmacokinetics of protein drugs. Ion-exchange chromatography (IEX) is among the most frequently used purification methods for PEGylated proteins. However, the underlying physical mechanisms allowing for a separation of different PEGamers (proteins with a varying number of attached PEG molecules) are not yet fully understood. In this work, mechanistic chromatography modeling was applied to gain a deeper understanding of the mass transfer and adsorption/desorption mechanisms of mono-PEGylated proteins in IEX...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731568/a-strong-hybrid-fatty-acid-inducible-transcriptional-sensor-built-from-yarrowia-lipolytica-upstream-activating-and-regulatory-sequences
#14
Murtaza Shabbir-Hussain, Ian Wheeldon, Mark A Blenner
The engineering of Yarrowia lipolytica to accumulate lipids with high titers and productivities has been enabled with a handful of constitutive promoters for pathway engineering. However, the development of promoters that are both strong and lipid responsive could greatly benefit the bioproduction efficiency of lipid-derived oleochemicals in oleaginous yeast. in this study, we sought to engineer a fatty acid regulated hybrid promoter for use in Y. lipolytica. We identified a 200 bp upstream regulatory sequence in the peroxisomal acyl CoA oxidase 2 (POX2) promoter...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731563/improving-fructose-utilization-and-butanol-production-by-clostridium-acetobutylicum-via-extracellular-redox-potential-regulation-and-intracellular-metabolite-analysis
#15
Li-Jie Chen, You-Duo Wu, Chuang Xue, Feng-Wu Bai
Jerusalem artichoke (JA) can grow well in marginal lands with high biomass yield, and thus is a potential energy crop for biorefinery. The major biomass of JA is from tubers, which contain inulin that can be easily hydrolyzed into a mixture of fructose and glucose, but fructose utilization for producing butanol as an advanced biofuel is poor compared to glucose-based ABE fermentation by Clostridium acetobutylicum. In this article, the impact of extracellular redox potential (ORP) on the process was studied using a mixture of fructose and glucose to simulate the hydrolysate of JA tubers...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731533/improvement-of-d-lactic-acid-production-in-saccharomyces-cerevisiae-under-acidic-conditions-by-evolutionary-and-rational-metabolic-engineering
#16
Seung-Ho Baek, Eunice Y Kwon, Sang-Jeong Bae, Bo-Ram Cho, Seon-Young Kim, Ji-Sook Hahn
Microbial lactic acid (LA) production under acidic fermentation conditions is favorable to reduce the production cost, but circumventing LA toxicity is a major challenge. We generated D-LA-producing Saccharomyces cerevisiae strain JHY5610 by expressing D-lactate dehydrogenase gene (Lm. ldhA) from Leuconostoc mesenteroides, while deleting genes involved in ethanol production (ADH1, ADH2, ADH3, ADH4, and ADH5), glycerol production (GPD1 and GPD2), and degradation of D-LA (DLD1). Adaptive laboratory evolution of JHY5610 led to a strain JHY5710 having higher LA tolerance and D-LA-production capability...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731532/formate-and-nitrate-utilization-in-enterobacter-aerogenes-for-semi-anaerobic-production-of-isobutanol
#17
Hwi-Min Jung, Yong Hwan Kim, Min-Kyu Oh
Anaerobic bioprocessing is preferred because of its economic advantages. However, low productivity and decreased growth of the host strain have limited the use of the anaerobic process. Anaerobic respiration can be applied to anoxic processing using formate and nitrate metabolism to improve the productivity of value-added metabolites. We constructed isobutanol-producing strains using Enterobacter aerogenes as a host strain by metabolic engineering approaches. The byproduct pathway (ldhA, budA, and pflB) was knocked out, and heterologous keto-acid decarboxylase (kivD) and alcohol dehydrogenase (adhA) were expressed along with the L-valine synthesis pathway (ilvCD and budB)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731528/improving-succinate-productivity-by-engineering-a-cyanobacterial-co2-concentrating-system-ccm-in-escherichia-coli
#18
Mengyong Xiao, Xinna Zhu, Changhao Bi, Yanhe Ma, Xueli Zhang
Biologically fixation of CO2 has great potential as a significant carbon source for biosynthesis, which is also a major way to reduce CO2 accumulation in atmosphere. Phosphoenolpyruvate (PEP) carboxylation is the key step of anaerobic succinate production in Escherichia coli. In this reaction, one mole CO2 is assimilated with PEP to form oxaloacetate by PEP carboxykinase (PCK). The preferred substrate of PCK is CO2 , which is very limited in cytoplasm. In this study, the carbon concentration mechanism (CCM) of cyanobacteria was introduced into E...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731527/mechanotransduction-effects-on-endothelial-cell-proliferation-via-cd31-and-vegfr2-implications-for-immunomagnetic-separation
#19
Kalpesh D Mahajan, Gauri M Nabar, Wei Xue, Mirela Anghelina, Nicanor Moldovan, Jeffrey Chalmers, Jessica Winter
Immunomagnetic separation is used to isolate circulating endothelial cells (ECs) and endothelial progenitor cells (EPCs) for diagnostics and tissue engineering. However, potentially detrimental changes in cell properties have been observed post-separation. Here, we studied the effect of mechanical force, which is naturally applied during immunomagnetic separation, on proliferation of human umbilical vein endothelial cells (HUVEC), kinase insert domain-positive receptor (KDR) cells, and peripheral blood mononuclear cells (PBMCs)...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28731525/differences-in-stem-cell-processing-lead-to-distinct-secretomes-secretion-implications-for-differential-results-of-previous-clinical-trials-of-stem-cell-therapy-for-myocardial-infarction
#20
Bernhard Wernly, Inês Gonçalves, Attila Kiss, Vera Paar, Tobias Mösenlechner, Michael Leisch, David Santer, Lucas Motloch, Klaus Ulrich Klein, Eva Verena Tretter, Daniel Kretzschmar, Bruno Podesser, Christian Jung, Uta C Hoppe, Michael Lichtenauer
INTRODUCTION: Stem cell therapy for acute myocardial infarction (AMI) seemed to be a promising therapy, however large clinical trials brought differential outcome. It has been shown that paracrine effects of secretomes of stem cells rather than cell therapy might play a fundamental role. The present study sought to compare cell processing protocols of clinical trials and investigated effects of differential cell culture conditions on chemokine secretion and functional effects. METHODS: Different secretomes were compared regarding IL-8, VEGF, MCP-1 and TNF-alpha secretion...
July 21, 2017: Biotechnology Journal
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