Abstract：
:This study addresses the problem of poor expression of somatotropin (ST) gene in E. coli and describes expression enhancement through silent and non-silent gene modifications. A series of constructs with codon optimization, substitution, deletion or addition in the 5'-region of the sequence encoding bubaline ST (BbST) were prepared. In the native form, the BbST expression was barely discernible on SDS-gel of the total E. coli cellular proteins (TCP). Introduction of silent and non-silent mutations in +2 to +8 codons, however, raised the expression levels to varying extents. In some constructs, a single base variation, i.e., G-->A or G-->C led to a remarkable increase in BbST expression (up to 28% of the TCP) whereas in the case of G-->T substitution the expression dropped to undetectable levels. Deletion of native GCC codon and addition of CAUCAC repeat thrice at +2 position enhanced the expression up to 48%, while insertion of NGG codons at the same position caused just a modest increase in expression. Differences in expression appeared as if related to the nature of early downstream codons (especially +2) and the stability of mRNA secondary structure although the levels of intracellular mRNA pools, as analyzed by real-time RT-PCR were quite similar. Overall, the study highlights the importance of 5'-end codon adaptations in solving the problems encountered in expressing the eukaryotic genes in E. coli.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Sadaf S,Khan MA,Akhtar MWdoi
10.1016/j.jbiotec.2008.03.010subject
Has Abstractpub_date
2008-06-01 00:00:00pages
134-9issue
2eissn
0168-1656issn
1873-4863pii
S0168-1656(08)00104-1journal_volume
135pub_type
杂志文章abstract：:Myxobacteria belonging to the genus Sorangium are known to produce a variety of biologically active secondary metabolites. Chivosazol is a macrocyclic antibiotic active against yeast, filamentous fungi and especially against mammalian cells. The compound specifically destroys the actin skeleton of eucaryotic cells and...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.10.011
更新日期：2006-01-24 00:00:00
abstract：:Aliphatic thiols are effective as redox buffers for folding non-native disulfide-containing proteins into their native state at high pH values (8.0-8.5) but not at neutral pH values (6-7.5). In developing more efficient and flexible redox buffers, a series of aromatic thiols was analyzed for its ability to fold scramb...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2004.09.005
更新日期：2005-02-09 00:00:00
abstract：:The present work describes the development of a novel direct screening method, assayed in 96-well format, for evaluation of enzymatic alkyl glycoside production in a hexanol-water two-phase system. Alkyl glycosides are surfactants with a range of applications and with good biodegradability and low toxicity. Enzymatic ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.11.005
更新日期：2010-01-15 00:00:00
abstract：:Structural composition and gene content of a biogas-producing microbial community from a production-scale biogas plant fed with renewable primary products was recently analyzed by means of a metagenome sequencing approach. To determine the transcriptionally active part of the same biogas community and to identify key ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.01.020
更新日期：2012-04-30 00:00:00
abstract：:An expression vector has been generated using a gene highly expressed under conditions found in a typical fed-batch bioreactor process. The ferritin heavy chain (HC) gene exhibits higher levels of expression in the late stages of a fed-batch bioreactor than in the early stages. This property was considered advantageou...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.09.021
更新日期：2007-01-30 00:00:00
abstract：:Terminal sialic acid on oligosaccharides of glycoproteins shows several biological functions of the glycoproteins. The yeast Pichia pastoris normally does not contain sialic acid on the oligosaccharides of glycoproteins. A sialyltransferase (ST) gene was transfected into P. pastoris to assess the possibility of using ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(00)00268-6
更新日期：2000-07-28 00:00:00
abstract：:Monitoring cell growth is crucial to the success of an animal cell culture process that can be accomplished by a variety of direct or indirect methodologies. Glucose is a major carbon and energy source for cultured mammalian cells in most cases, but glycolytic metabolism often results in the accumulation of lactate. G...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.05.016
更新日期：2005-08-22 00:00:00
abstract：:Streptomyces globisporus C-1027 produces a nine-membered enediyne antitumor antibiotic lidamycin. Here we report the complete genome sequence of S. globisporus C-1027, which consists of a 7,608,611 bp linear chromosome, a 167,754 bp linear plasmid SGLP1 and a 7,234 bp circular plasmid pSGL1. The biosynthetic gene clus...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.02.004
更新日期：2016-03-20 00:00:00
abstract：:The methylotropic yeast Pichia pastoris has demonstrated its suitability for large-scale production of recombinant proteins. As an eukaryotic organism P. pastoris presents a series of advantages at expression and processing of heterologous proteins when compared with Escherichia coli. In this work, P. pastoris has bee...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.04.024
更新日期：2016-06-10 00:00:00
abstract：:Zeolitic imidazole frameworks (ZIFs) with tunable pore sizes and high surface areas have recently used as an effective support for immobilizing enzymes. However, the instability in the aqueous acidic environment has limited their practical applications in some cases. In this work, we develop a novel catalase/ZIFs comp...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.09.010
更新日期：2019-12-20 00:00:00
abstract：:Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in ext...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.10.015
更新日期：2010-01-15 00:00:00
abstract：:There has been little research reported on the synergism of plant expansins in cellulose hydrolysis because of the difficulty of heterologous expression. In this study, the expansin gene LeExp2 of Lycopersicum esculentum was expressed for the first time in Pichia pastoris to identify its function in vitro. The optimal...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.07.013
更新日期：2014-09-30 00:00:00
abstract：:We introduce an easy, fast and effective method to analyze the influence of genetically modified (GM) plants on soil and model organisms in the laboratory to substitute laborious and time consuming field trials. For the studies described here we focused on two GM plants of the so-called 3rd generation: GM plants produ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.07.012
更新日期：2017-09-10 00:00:00
abstract：:To develop a receptor-mediated intracellular delivery system that can transport therapeutic proteins to specific tumor cells, folate-poly(ethylene glycol)-grafted-trimethylchitosan (folate-PEG-g-TMC) was synthesized. Nano-scaled spherical polyelectrolyte complexes between the folate-PEG-g-TMC and fluorescein isothiocy...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.09.007
更新日期：2010-01-01 00:00:00
abstract：:Laccase from the white rot fungus Coriolopsis polyzona was immobilized for the first time through the formation of cross-linked enzyme aggregates (CLEAs). Laccase CLEAs were produced by using 1000g of polyethylene glycol per liter of enzyme solution as precipitant and 200muM of glutaraldehyde as a cross-linking agent....
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.07.948
更新日期：2007-10-15 00:00:00
abstract：:The catalytic efficiency and pH optimum of Trichoderma reesei endo-beta-1,4-glucanase II were improved by protein engineering. We subjected residue 342 to saturation mutagenesis, and further changed the enzyme by random mutagenesis and two rounds of DNA shuffling. Enzyme variants were purified and characterized. Varia...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2008.03.016
更新日期：2008-06-01 00:00:00
abstract：:An enzyme-responsive artificial chaperone system which employs an amphiphilic amylose primer (dodecyl maltopentaose, C12-MP) as a surfactant and phosphorylase b was designed to enable protein refolding. Effective refolding of carbonic anhydrase B after both heat denaturation (70 degrees C for 10min) and guanidine hydr...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.01.013
更新日期：2009-03-25 00:00:00
abstract：:Exploring novel biological anti-quorum sensing (QS) agents to control membrane biofouling is of great worth in order to allow sustainable performance of membrane bioreactors (MBRs) for wastewater treatment. In recent studies, QS inhibitors have provided evidence of alternative route to control membrane biofouling. Thi...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.06.029
更新日期：2012-10-31 00:00:00
abstract：:The study describes extraction of extracellular polymeric substances (EPS) from sewage sludge by applying enzymes and enzymes combined with sodium tripolyphosphate (STPP). Additionally, a systematic study of two non-enzymatic extraction agents is described. The assessment of the released products is made by colorimetr...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.04.005
更新日期：2007-06-30 00:00:00
abstract：:Streptomyces lividans TK24 is the standard host for the heterologous expression of a number of different proteins and antibiotic-synthesizing enzymes. As such, it is often used as an experimental microbial cell factory for the production of secreted heterologous proteins including human cytokines and industrial enzyme...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.02.004
更新日期：2015-04-10 00:00:00
abstract：:A structured kinetic model that accounts for proteolytic degradation due to recombinant protein overexpression is introduced and its performance evaluated by comparison with previously reported fed-batch experimental data. This mathematical model contains an additional pool for a generic key precursor (in our case phe...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(99)00014-0
更新日期：1999-05-28 00:00:00
abstract：:4-vinylguaiacol, a kind of volatile phenolic compound with tobacco flavor, is widely used as a component of edible flavor and intermediate of medicine. Ferulic acid is usually used as substrate for the biosynthesis of 4-vinylguaiacol. However, the price of ferulic acid is high, leading to high production cost. In this...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2018.07.021
更新日期：2018-09-10 00:00:00
abstract：:In this article, we review how molecular modeling techniques can be used to shed light on how water and organic solvents influence the reactivity of enzymes. The application of thermodynamics-based models allowed the first qualitative predictions on the selectivity of many reaction types. However, it was with the appl...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/s0168-1656(02)00034-2
更新日期：2002-06-13 00:00:00
abstract：:Candida antarctica lipase B (CALB) has been employed as an efficient catalyst in the preparation of many flavor esters. A CALB-displaying yeast whole-cell biocatalyst could be an attractive alternative to commercial immobilized CALB because of its low-cost preparation and high enzymatic activity. We investigated the p...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2012.02.013
更新日期：2012-05-31 00:00:00
abstract：:Biosurfactants are surface-active agents produced by microorganisms and show increasing significance in various industrial applications. A great variety of these secondary metabolites are described to occur within actinomycetes, amongst trehalose lipids and oligosaccharide lipids produced by the family Tsukamurellacea...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.06.424
更新日期：2015-10-10 00:00:00
abstract：:A simple and accurate analysis system for the estimation of recombination efficiency in vivo using fluorescence-activated cell sorting (FACS) was designed and was subsequently used to compare the efficiency of recombination related to different spacer mutants. F(3) and F(5) mutant sequences were used for Flpe-mediated...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.07.027
更新日期：2007-01-10 00:00:00
abstract：:Anaerobic hydrocarbon biodegradation not only diminishes fuel quality, but also exacerbates the biocorrosion of the metallic infrastructure. While successional events in marine microbial ecosystems impacted by petroleum are well documented, far less is known about the response of communities chronically exposed to hyd...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.02.021
更新日期：2017-08-20 00:00:00
abstract：:Lentinula edodes, the popular shiitake mushroom, is one of the most important cultivated edible mushrooms. It is used as a food and for medicinal purposes. Here, we present the 46.1 Mb draft genome of L. edodes, comprising 13,028 predicted gene models. The genome assembly consists of 31 scaffolds. Gene annotation prov...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.02.032
更新日期：2016-04-10 00:00:00
abstract：:Several green fluorescent protein (Gfp) mutants with increased cellular fluorescence compared to the wildtype protein have recently been generated. We have expressed and compared wildtype Gfp and mutants S65T, F100S/ M154T/V164A, F64L/S65T, and S65A/V68L/S72A under identical growth conditions in CHO and Saccharomyces ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(98)00040-6
更新日期：1998-06-11 00:00:00
abstract：:We have reported that nanowell array (NWA) can enhance electrochemical detection of molecular binding events by controlling the binding sites of the captured molecules. Using NWA biosensor based amperometric analysis, we have detected biological macromolecules such as DNA, protein or aptamers at low concentrations. In...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2013.08.029
更新日期：2013-12-01 00:00:00
