Abstract:
:Gene deletion strategies using flux balance analysis (FBA) have improved the growth-coupled production of various compounds. However, the productivities were often below the expectation because the cells failed to adapt to these genetic perturbations. Here, we demonstrate the productivity of the succinate of the designed gene deletion strain was improved by adaptive laboratory evolution (ALE). Although FBA predicted deletions of adhE-pykAF-gldA-pflB lead to produce succinate from glycerol with a yield of 0.45 C-mol/C-mol, the knockout mutant did not produce only 0.08 C-mol/Cmol, experimentally. After the ALE experiments, the highest succinate yield of an evolved strain reached to the expected value. Genome sequencing analysis revealed all evolved strains possessed novel mutations in ppc of I829S or R849S. In vitro enzymatic assay and metabolic profiling analysis revealed that these mutations desensitizing an allosteric inhibition by L-aspartate and improved the flux through Ppc, while the activity of Ppc in the unevolved strain was tightly regulated by L-aspartate. These result demonstrated that the evolved strains achieved the improvement of succinate production by expanding the flux space of Ppc, realizing the predicted metabolic state by FBA.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Tokuyama K,Toya Y,Horinouchi T,Furusawa C,Matsuda F,Shimizu Hdoi
10.1002/bit.26568subject
Has Abstractpub_date
2018-06-01 00:00:00pages
1542-1551issue
6eissn
0006-3592issn
1097-0290journal_volume
115pub_type
杂志文章abstract::A mathematical model is formulated to describe trends in biomass and penicillin formation as well as substrate consumption for fed-batch cultivations. The biomass is structured into three morphological compartments, and glucose and corn steep liquor are considered as substrates for growth. Penicillin formation is assu...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(SICI)1097-0290(19971220)56:6<593::AID-BIT
更新日期:1997-12-20 00:00:00
abstract::An integrated approach is used to develop a rapid sampling strategy for the quantitative analysis of in vivo kinetic behavior based on measured concentrations of intracellular metabolites in Saccharomyces cerevisiae. Emphasis is laid on small sample sizes during sampling and analysis. Subsecond residence times are acc...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10048
更新日期:2001-11-20 00:00:00
abstract::The fermentation kinetics of methane production from whey permeate in a packed bed immobilized cell bioreactor at mesophilic temperatures and pHs around neutral was studied. Propionate and acetate were the only two major organic intermediates found in the methanogenic fermentation of lactose. Based on this finding, a ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260370412
更新日期:1991-02-20 00:00:00
abstract::Being biosurfactants, rhamnolipids create severe foaming when produced in aerobic Pseudomonas aeruginosa fermentation. The necessary reduction of aeration causes oxygen limitation and restricts cell and product concentrations. In this study, we evaluate the new strategy of rhamnolipid production under denitrification ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/1097-0290(20010105)72:1<25::aid-bit4>3.0.c
更新日期:2001-01-05 00:00:00
abstract::The microfluidic environment provided by implanted prostheses has a decisive influence on the viability, proliferation and differentiation of cells. In bone tissue engineering, for instance, experiments have confirmed that a certain level of wall shear stress (WSS) is more advantageous to osteoblastic differentiation....
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22842
更新日期:2010-11-01 00:00:00
abstract::Anerobic biofilms with dominantly acidogenic bacteria were grown in fixed-bed recycle reactors. The influence of calcium concentration in the culture medium on biofilm mass accumulation, immobilized calcium concentration, and biofilm-specific activity was investigated. The results indicate that the biofilm mass accumu...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260450305
更新日期:1995-02-05 00:00:00
abstract::The influence of solution Eh on the rate of ferrous iron oxidation by Acidithiobacillus ferrooxidans is characterized. The experimental approach was based on the use of a two-chamber bioelectrochemical cell, which can determine the ferrous iron oxidation rate at controlled potential. Results enabled the formulation of...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10371
更新日期:2002-11-05 00:00:00
abstract::10-Deacetylabaccatin III (10 DAB), an important precursor for paclitaxel semisynthesis, is enhanced in yew extracts using C10-deacetylase and C13-deacylase enzymes.(4) C10-deacetylase is an intracellular enzyme produced by the fermentation of a soil microorganism, Nocardioides luteus (SC 13912). During the fermentatio...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260480518
更新日期:1995-12-05 00:00:00
abstract::Modern large-scale agricultural practices that incorporate high density farming with subtherapeutic antibiotic dosing are considered a major contributor to the rise of antibiotic-resistant bacterial infections of humans with species of Salmonella being a leading agriculture-based bacterial infection. Microcin J25, a p...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.26772
更新日期:2018-10-01 00:00:00
abstract::In view of the biochemical reaction catalyzed by enzyme powder suspended in a water-insoluble organic solvent, an equation was derived to estimate the amount of water bound to the enzyme powder. With this equation, an apparent adsorption isotherm between free water (water freely dissolved in benzene) and bound water (...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260340613
更新日期:1989-09-01 00:00:00
abstract::Product inhibition of biological production systems is a widely observed phenomenon with prominent implications for the design and ultimately the success of biotechnological processes. In order to investigate whether such effects could limit the maximal concentrations in the production of the beta-lactamase inhibitor ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21783
更新日期:2008-06-15 00:00:00
abstract::Saccharomyces cerevisiae (yeast) cells were employed as a source of alcohol dehydrogenase in the NAD(+)-to-NADH reaction. The cells were immobilized in calcium alginate monofilament fibers and used in a biological reactor. The alginate could not be heat sterilized since temperatures above 80 degrees C caused the polym...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260360706
更新日期:1990-10-05 00:00:00
abstract::Protein phosphorylation is an important post-translational modification for intracellular signaling molecules, mostly found in serine and threonine residues. Tyrosine phosphorylations are very few events (less than 0.1% to phosphorylated serine/threonine residues), but capable of governing cell fate decisions involved...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.25948
更新日期:2016-08-01 00:00:00
abstract::This research investigates the effect of solids retention time (SRT) on the acid-phase anaerobic digestion of primary sludge. A series of experiments were conducted using two continuous-flow 3-L units with the following configuration: a completely mixed reactor (CMR) with clarifier and solids recycle and an upflow ana...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260440103
更新日期:1994-06-05 00:00:00
abstract::Advanced cell and gene therapies such as chimeric antigen receptor T-cell immunotherapies (CAR-T), present a novel therapeutic modality for the treatment of acute and chronic conditions including acute lymphoblastic leukemia and non-Hodgkin lymphoma. However, the development of such immunotherapies requires the manufa...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.27088
更新日期:2019-10-01 00:00:00
abstract::An immobilized enzyme reactor has been developed for the degradation of bilirubin as a potential treatment for neonatal jaundice. It utilizes the enzyme bilirubin oxidase from Myrothecium verrucaria, which in the presence of molecular oxygen converts bilirubin to biliverdin and other products that are much less toxic ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260281011
更新日期:1986-10-01 00:00:00
abstract::The production of acetate by aerobically growing E. coli is examined. The problem is formulated in terms of a flow network that has as its objective maximal ATP synthesis. It is found that when loads are imposed and flux constraints exist either at the level of NADH turnover rate or the activity of a key Krebs cycle e...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260350711
更新日期:1990-03-25 00:00:00
abstract::Pronounced spatial nonuniformities in cell density, physiology, and activity frequently arise within densely packed immobilized cell supports. For a more fundamental understanding of immobilized cell phenomena, we have developed high-resolution microfluorimetric procedures to analyze local variations in both immobiliz...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260380404
更新日期:1991-08-05 00:00:00
abstract::The high hydrophobicity of polycyclic aromatic hydrocarbons (PAHs) strongly reduces their bioavailability in aged contaminated soils, thus limiting their bioremediation. The biodegradation of PAHs in soils can be enhanced by employing surface-active agents. However, chemical surfactants are often recalcitrant and exer...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.20225
更新日期:2004-10-20 00:00:00
abstract::Alginate microparticles produced by emulsification/internal gelation were investigated as a promising carrier for insulin delivery. The procedure involves the dispersion of alginate solution containing insulin protein, into a water immiscible phase. Gelation is triggered in situ by instantaneous release of ionic calci...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21164
更新日期:2007-04-01 00:00:00
abstract::A mathematical model is developed to analyze the influence of chemical reaction and diffusion processes on the intracellular organization of mitochondria in skeletal muscle cells. The mathematical modeling approach uses a reaction-diffusion analysis of oxygen, ATP, and ADP involved in energy metabolism and mitochondri...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.23112
更新日期:2011-08-01 00:00:00
abstract::A simple methodology has been successfully employed to explain the solvent dependence of the substrate specificity of enzymes in organic media. This methodology, which does not require the knowledge of the enzyme structure and is thus applicable to lyophilized and other noncrystalline enzyme preparations, predicts tha...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(SICI)1097-0290(19971105)56:3<340::AID-BIT
更新日期:1997-11-05 00:00:00
abstract::The biotransformation of benzaldehyde and glucose to L-phenylacetylcarbinol (PAC) using Candida utilis was demonstrated in a solid-liquid two-phase partitioning bioreactor (TPPB) with the aim of reducing substrate, product, and by-product toxicity via sequestration. Previous work in the field had used octanol as the s...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22839
更新日期:2010-11-01 00:00:00
abstract::Fungal morphology in many filamentous fungal fermentations leads to high broth viscosity which limits oxygen mass transfer, and often results in reduced productivity. The objective in this study was to determine if a simple, fed-batch, process strategy-pulsed addition of limiting-carbon source-could be used to reduce ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10548
更新日期:2003-04-05 00:00:00
abstract::A method was developed for obtaining high signal-to-noise 13C NMR spectra of intracellular compounds in metabolically active cultured cells. The method allows TCA cycle labeling kinetics to be determined in real time without significant oxygen transport limitations. Cells were immobilized on the surface of nonporous m...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.20191
更新日期:2004-09-30 00:00:00
abstract::The diffusion of viruses toward cells is a limiting step of the infection process. To be modeled correctly, this step must be evaluated in combination with the adsorption of the virus to the cell surface, which is a rapid but reversible step. In this paper, the recombinant adenovirus (rAd) diffusion and its adsorption...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.21340
更新日期:2007-09-01 00:00:00
abstract::The emissions of nitrous oxide (N(2)O) and nitric oxide (NO) from biological nitrogen removal (BNR) operations via nitrification and denitrification is gaining increased prominence. While many factors relevant to the operation of denitrifying reactors can influence N(2)O and NO emissions from them, the role of differe...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22704
更新日期:2010-06-15 00:00:00
abstract::Alcoholic fermentation of cheese whey permeate was investigated using a recombinant flocculating Saccharomyces cerevisiae, expressing the LAC4 (coding for beta-galactosidase) and LAC12 (coding for lactose permease) genes of Kluyveromyces marxianus enabling for lactose metabolization. Data on yeast fermentation and gro...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/1097-0290(20010305)72:5<507::aid-bit1014>3
更新日期:2001-03-05 00:00:00
abstract::The copolymerization method of immobilization was used to obtain preparations of enzymes covalently incorporated in polyacrylamide gel. They possess properties making them suitable for practical use. First, the preparations are hundreds of times more stable against irreversible thermoinactivation than native enzymes. ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260250804
更新日期:1983-08-01 00:00:00
abstract::This article studies the operation of a new process for the production of biopolymers (polyhydroxyalkanoates, PHAs) at different applied organic load rates (OLRs). The process is based on the aerobic enrichment of activated sludge to obtain mixed cultures able to store PHAs at high rates and yields. A mixture of aceti...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.20683
更新日期:2006-01-05 00:00:00