Abstract：
:The expression of recombinant yeast pyruvate carboxylase (PYC) in animal cell lines was shown in previous studies to reduce significantly the formation of waste metabolites, although it has translated into mixed results in terms of improved cellular growth and productivity. In this work, we demonstrate that the unique phenotype of PYC expressing cells can be exploited through the application of a dynamic fed-batch strategy and lead to significant process enhancements. Metabolically engineered HEK293 cells stably producing human recombinant IFNα2b and expressing the PYC enzyme were cultured in batch and fed-batch modes. Compared to parental cells, the maximum cell density in batch was increased 1.5-fold and the culture duration was extended by 2.5 days, but the product yield was only marginally increased. Further improvements were achieved by developing and implementing a dynamic fed-batch strategy using a concentrated feed solution. The feeding was based on an automatic control-loop to maintain a constant glucose concentration. This strategy led to a further 2-fold increase in maximum cell density (up to 10.7×10(6)cells/ml) and a final product titer of 160mg/l, representing nearly a 3-fold yield increase compared to the batch process with the parental cell clone.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Vallée C,Durocher Y,Henry Odoi
10.1016/j.jbiotec.2013.11.002subject
Has Abstractpub_date
2014-01-01 00:00:00pages
63-70eissn
0168-1656issn
1873-4863pii
S0168-1656(13)00485-9journal_volume
169pub_type
杂志文章abstract：:The aim of the present study was to investigate the effect of three routine drug excipients, as osmolytes, in three different concentrations, on structure, thermal stability and the activity of single-chain (sc-) tenecteplase. To see the influence of trehalose, mannitol, and sucrose on the structure, stability and func...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.05.014
更新日期：2018-08-20 00:00:00
abstract：:A transgenic tobacco plant (Nicotiana tabacum L.) expressing a fungal pectin methylesterase (PME; EC 3.1.1.11) gene derived from a black filamentous fungus, Aspergillus niger was created. Fungal PME should have a wider range of adaptability to substrate pectin compared with plant PME. As expected, the proportion of me...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.04.015
更新日期：2004-08-05 00:00:00
abstract：:Randomly amplified polymorphic DNA (RAPD) was used to analyzed 78 samples comprises of certified reference materials (soya and maize powder), raw seeds (soybean and maize), processed food and animal feed. Combination assay of two arbitrary primers in the RAPD analysis enable to distinguish genetically modified organis...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.06.001
更新日期：2006-12-15 00:00:00
abstract：:Increasing levels of antibiotic resistance in pathogens, including Staphylococcus aureus, remains a serious problem for public health, leading to the need for better alternative antimicrobial strategies. The antimicrobial proteins produced by Lactobacillus plantarum USM8613 attributed to its anti-staphylococcal activi...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.05.006
更新日期：2019-07-20 00:00:00
abstract：:The poly(beta-hydroxybutyrate) (PHB) biosynthetic genes of Ralstonia eutropha that are organized in a single operon (phaCAB) have been cloned in Escherichia coli, where the expression of the genes in the wild-type pha operon from plasmid pTZ18U-PHB leads to the formation of 50-80% PHB/celldry mass when the cells are g...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(98)00127-8
更新日期：1998-10-27 00:00:00
abstract：:The marine microalga Nannochloropsis oculata is a potential source of eicosapentaenoic acid (EPA, C20:5n3) and carotenoids for use in functional foods and nutraceuticals. Mixotrophic culture of N. oculata using glycerol was examined as a possible way of increasing the biomass and metabolite productivity relative to a ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.10.006
更新日期：2016-12-10 00:00:00
abstract：:CRISPR-Cpf1 is a type V CRISPR system that has recently been exploited for genome engineering purposes. Compared to the well-known Streptococcus pyogenes CRISPR-Cas9 system, the effector protein Cpf1 recognizes T-rich protospacer-adjacent motif (PAM) instead of G-rich PAM (used by CRISPR-Cas9), which could offer a sub...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.07.040
更新日期：2018-10-20 00:00:00
abstract：:Glutamate dehydrogenase is a central enzyme connecting nitrogen and carbon metabolism via its precursors ammonium and oxoglutarate and its product glutamate. In Corynebacterium glutamicum glutamate dehydrogenase is especially important, since it is a key enzyme for the biotechnological production of the flavour enhanc...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.04.007
更新日期：2009-06-15 00:00:00
abstract：:The pyranose 2-oxidases from Trametes ochracea and Trametes pubescens share markedly similar amino acid sequences with identity of 93.4%. When expressed from the recombinant plasmids based on the same vector in the Escherichia coli host strain BL21(DE3) at higher growth temperatures, they differ strikingly in the form...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.04.023
更新日期：2007-06-30 00:00:00
abstract：:Expression of cloned PhiX174 gene E in bacteria results in lysis of bacteria. It is unique among phage lysis systems as it introduces a transmembrane tunnel structure through the cell envelope complex of Gram-negative bacteria. The resulting bacterial ghosts have intact envelope structures devoid of cytoplasmic conten...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/0168-1656(95)00123-9
更新日期：1996-01-26 00:00:00
abstract：:Melanins are predominantly indolic polymers which are having extensive applications in cosmetics, agriculture and medicine. In the present study, optimization of nutritional parameters influencing melanin production by Mucuna monosperma callus cultures was attempted using the response surface methodology (RSM). Standa...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2013.11.012
更新日期：2014-01-20 00:00:00
abstract：:Oleaginous microalgae have received a considerable attention as potential biofuel feedstock. However, lack of industry-suitable strain with lipid rich biomass limits its commercial applications. Targeted engineering of lipogenic pathways represents a promising strategy to enhance the efficacy of microalgal oil product...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.05.005
更新日期：2016-07-10 00:00:00
abstract：:An alkalophilic bacterium was isolated based on the potential of extra-cellular enzymes for bioscouring. The bacterium was identified as a new strain of Bacillus pumilus BK2 producing an extra-cellular endo-pectate lyase PL (EC 4.2.2.2). PL was purified to homogeneity in three steps and has a molecular mass of 37.3+/-...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.07.019
更新日期：2006-02-10 00:00:00
abstract：:The aim of this work was the production of bioactive metabolites by submerged fermentation from the fungus Diaporthe schini, followed by their extraction, separation and characterization. Different solvents (methanol, dichloromethane and hexane) were used for the extraction of metabolites from the fermentation broth a...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.01.022
更新日期：2019-03-20 00:00:00
abstract：:During the course of a lifetime, it has been calculated that we may consume between 100 and 700 tons of food. For the average British citizen, this is likely to include some 550 poultry, 36 pigs, 36 sheep, eight oxen, 10000 eggs and dairy products (milk, butter cheese, etc.) equivalent to 18 tonnes of milk. As if that...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/s0168-1656(00)00299-6
更新日期：2000-09-29 00:00:00
abstract：:The invariant surface glycoprotein ISG75 is a transmembrane glycoprotein occurring on the surface of the bloodstream-form Trypanozoon. This study describes the expression and purification of the N-terminal extracellular domain of ISG75, a novel target for development of diagnostic tests for trypanosomosis. To facilita...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.04.012
更新日期：2008-06-30 00:00:00
abstract：:The first complete genome sequence of Bacillus glycinifermentans B-27 was determined by SMRT sequencing generating a genome sequence with a total length of 4,607,442 bases. Based on this sequence 4738 protein-coding sequences were predicted and used to identify gene clusters that are related to the production of secon...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.04.021
更新日期：2017-09-10 00:00:00
abstract：:Enteric infections associated with bacteria, viruses and parasites are a world health problem accounting for at least one billion episodes of diarrhoea and up to 10 million deaths in the Third World each year. The widely held conclusion that responses which are protective in the gastrointestinal tract are best stimula...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/0168-1656(95)00114-X
更新日期：1996-01-26 00:00:00
abstract：:Phycocyanin (PC) prepared from a cyanobacterium Spirulina platensis by the DEAE-DE52 cellulose column chromatography that was developed by gradient elution of 50-250 mM phosphate buffer (pH 7.0) was stabilized by its subunits cross-linked covalently with formaldehyde. The single blue band that the chemically stabilize...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.08.017
更新日期：2006-02-24 00:00:00
abstract：:Recombinant Escherichia coli clones encoding between 0 and 6 copies of the mammalian cytochrome b5 gene were subjected to pyrolysis mass spectrometry (PyMS). To deconvolute the pyrolysis mass spectra so as to obtain quantitative information on the amount of cytochrome b5 produced fully-interconnected feedforward artif...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(94)90088-4
更新日期：1994-05-15 00:00:00
abstract：:We cloned and characterized a preprochymosin cDNA from the abomasum of milk-fed kid goats. This cDNA contained an open reading frame that predicts a polypeptide of 381 amino acid residues, with a signal peptide and a proenzyme region of 16 and 42 amino acids, respectively. Comparison of the caprine preprochymosin sequ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.06.002
更新日期：2004-10-19 00:00:00
abstract：:The efficiency of existing combinatorial biological library methods has been moderate in terms of the success rates, the affinities of the ligands selected and the time and effort involved in trying to optimize the initial leads. Although mimicking natural evolution, existing strategies take little notice of the impor...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/s1389-0352(01)00019-8
更新日期：2001-06-01 00:00:00
abstract：:Paenibacillus riograndensis is a Gram-positive rhizobacterium which exhibits plant growth promoting activities. It was isolated from the rhizosphere of wheat grown in the state of Rio Grande do Sul, Brazil. Here we announce the complete genome sequence of P. riograndensis strain SBR5(T). The genome of P. riograndensis...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.04.025
更新日期：2015-08-10 00:00:00
abstract：:Protein engineering has been employed to investigate the effect of specific amino acid changes on the targeting of heterologous proteins to the outer cell surface of the Gram-positive bacterium Staphylococcus xylosus. Three different variants, corresponding to a 101 amino acid region of the major glycoprotein (G prote...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(95)00081-z
更新日期：1995-10-16 00:00:00
abstract：:RNF2 is a core component of Polycomb repressive complex 1 (PRC1), which binds to various chromatic regions to regulate their transcriptional activity. It was recently reported that EZH2, a Polycomb group complex member, helps mediate long-range chromosomal interactions in mammalian cells. The present study investigate...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2011.01.001
更新日期：2011-02-20 00:00:00
abstract：:The hematopoietic system represents an interesting model for gene transfer protocols. Here, we have evaluated the efficiency of a gene transfer system using the polycationic compound SuperFect (Qiagen) and the K562 hematopoietic cell line. Transient and stable vectors carrying the enhanced green fluorescent protein (E...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(01)00276-0
更新日期：2001-06-15 00:00:00
abstract：:Human interleukin (IL)-29 is the latest member of the class II cytokine family. However, as a result of lacking efficient method to generate relatively large quantity of IL-29, little is known of its functions in man. In the present study, an Escherichia coli expression system for the rapid expression of the human IL-...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.11.019
更新日期：2006-04-10 00:00:00
abstract：:Protein modifications by intricate cellular machineries often redesign the structure and function of existing proteins to impact biological networks. Disulfide bond formation between cysteine (Cys) pairs is one of the most common modifications found in extracellularly-destined proteins, key to maintaining protein stru...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.03.006
更新日期：2017-04-20 00:00:00
abstract：:Rebuilding enzymes with high activities, stability and reusability has been constantly pursued for application of enzymaticcatalysis in wide field. Rebuilding enzymes by chemical approaches, including immobilization and chemicalmodification, represents a simple but effective route. Here, a class of highly stable and a...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.12.014
更新日期：2019-02-10 00:00:00
abstract：:In this work it is describe for the first time, the use of an esterase with null activity (Tfu_0883 bacterial cutinase from Thermobifida fusca) on the removal of fat from the surface of a cotton substrate. Similar levels of fat removal were found for both null and wild-type proteins, despite that only wild type protei...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.06.019
更新日期：2012-10-31 00:00:00
