Abstract：
:Fluorescence in situ hybridization (FISH) has been extensively used in the past decades for the detection and localization of microorganisms. However, a mechanistic approach of the whole FISH process is still missing, and the main limiting steps for the hybridization to occur remain unclear. In here, FISH is approached as a particular case of a diffusion-reaction kinetics, where molecular probes (MPs) move from the hybridization solution to the target RNA site within the cells. Based on literature models, the characteristic times taken by different MPs to diffuse across multiple cellular barriers, as well as the reaction time associated with the formation of the duplex molecular probe-RNA, were estimated. Structural and size differences at the membrane level of bacterial and animal cells were considered. For bacterial cells, the limiting step for diffusion is likely to be the peptidoglycan layer (characteristic time of 7.94 × 102 - 4.39 × 103 s), whereas for animal cells, the limiting step should be the diffusion of the probe through the bulk (1.8-5.0 s) followed by the diffusion through the lipid membrane (1 s). The information provided here may serve as a basis for a more rational development of FISH protocols in the future.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Lima JF,Maia P,T Magalhães B,Cerqueira L,Azevedo NFdoi
10.1002/bit.27462subject
Has Abstractpub_date
2020-10-01 00:00:00pages
3212-3223issue
10eissn
0006-3592issn
1097-0290journal_volume
117pub_type
杂志文章,评审abstract：:A two-member methane-utilizing mixed culture of bacteria, formed by combining two pure cultures isolated from a naturally occurring methane-utilizing mixed culture, was studied in continuous culture. From the nutritional requirements and substrate ranges of the pure cultures, a mechanism for the interspecific interact...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260221010
更新日期：1980-10-01 00:00:00
abstract：:Control of cell shape and behavior through the micropattern technique by spatial immobilization of adhesive proteins on a surface has provided novel insights in several aspects of cell biology, such as tissue morphogenesis, cell growth and cell differentiation, and apoptosis. In this work, we present the use of poly(e...
journal_title：Biotechnology and bioengineering
pub_type： 信件
doi：10.1002/bit.22997
更新日期：2011-04-01 00:00:00
abstract：:The analysis of a continuous, aerobic, fixed-film bioreactor is performed by simulating the behavior of penicillin production in a three-phase fluidized bed. Rigorous mathematical models are developed for a fluidized-bed fermentor in which bioparticles are fluidized by the liquid medium and air. The steady-state perfo...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260260509
更新日期：1984-05-01 00:00:00
abstract：:Extractive work-up of whole-cell biotransformations generally suffers from the formation of stable gels and slimes upon addition of the organic solvent to the cell suspension and the cell-free solution, respectively. This problem has been overcome by enzymatic lysis of emulsifying agents present in the medium through ...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20155
更新日期：2004-08-20 00:00:00
abstract：:A computer-controlled headspace gas chromatograph was used to monitor the progress of ethanol production from both aerobic batch and anaerobic continuous fermentations. Using an automatic, electropneumatic sampling system, aliquots of fermentation headspace gas were injected directly onto the column for quantitative e...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260251103
更新日期：1983-11-01 00:00:00
abstract：:A model that was used to describe toxicity from high concentrations of chlorinated aliphatic hydrocarbons (CAHs) on reductively dechlorinating cultures in batch reactors (Sabalowsky and Semprini (in press)) was extended here to simulate observations in continuous flow suspended and attached growth reactors. The reduct...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.22822
更新日期：2010-10-15 00:00:00
abstract：:A refolding strategy was described for on-column refolding of recombinant human interferon-gamma (rhIFN-gamma) inclusion bodies by expanded bed adsorption (EBA) chromatography. After the denatured rhIFN-gamma protein bound onto the cation exchanger of STREAMLINE SP, the refolding process was performed in expanded bed ...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20763
更新日期：2006-03-05 00:00:00
abstract：:Innovation in monoclonal antibody (mAb) production continues to be driven by cell engineering strategies to increase yield and improve product quality. In a previous study, to investigate the effectiveness of transporter overexpression strategies, we prepared a taurine transporter-overexpressing host cell line (DXB11/...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.24881
更新日期：2013-08-01 00:00:00
abstract：:A novel process has been developed which uses reversed micelles to isolate denatured protein molecules from each other and allows them to refold individually. These reversed micelles are aqueous phase droplets stabilized by the surfactant AOT and suspended in isooctane. By adjusting conditions such that only one prote...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260351002
更新日期：1990-04-25 00:00:00
abstract：:The simultaneous biodegradation of the nonionic surfactant Tween 20 (Tw20) and pentachlorophenol (PCP) by Sphingomonas chlorophenolica sp. Strain RA2 (RA2) was measured. As a sole substrate, Tw20 biodegradation was best described by the Contois kinetic model. During concurrent biodegradation of Tw20 and PCP, the biode...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20365
更新日期：2005-03-20 00:00:00
abstract：:It is shown that the mass transfer resistance can significantly distort the linearity of the Lineweaver-Burk plot of the kinetic data for a microbial culture which forms aggregates. For small flocs, the linearity of the Lineweaver-Burk plot is largely retained, but a different slope and intercept will be obtained comp...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260191204
更新日期：1977-12-01 00:00:00
abstract：:This paper describes construction of a nanosensory device for amplified detection of biologically important amines as chemical signals. The device was inspired by a biological signal transduction system, and was fabricated on an artificial cell membrane through self-organization of the molecular components, such as a ...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.22521
更新日期：2010-01-01 00:00:00
abstract：:Lipases are versatile biocatalists showing multiple applications in a wide range of biotechnological processes. The gene lipA coding for Lipase A from Bacillus subtilis was isolated by PCR amplification, cloned and expressed in Escherichia coli, Saccharomyces cerevisiae and Bacillus subtilis strains, using pBR322, YEp...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.10201
更新日期：2002-05-05 00:00:00
abstract：:Proteinaceous organic wastes are suitable substrates to produce high added-value products in anaerobic mixed-culture fermentations. In these processes, the stoichiometry of the biotransformation depends highly on operational conditions such as pH or feeding characteristics and there are still no tools that allow the p...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.27177
更新日期：2020-01-01 00:00:00
abstract：:Acetate accumulation is a common problem observed in aerobic high cell density Escherichia coli cultures. A previous report has hypothesized that the glyoxylate shunt is active in a low acetate producer, E. coli BL21, and inactive in a high acetate producer, JM109. To further investigate this hypothesis, we now develo...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：
更新日期：2000-05-05 00:00:00
abstract：:Rifamycin B is an important polyketide antibiotic used in the treatment of tuberculosis and leprosy. We present results on medium optimization for Rifamycin B production via a barbital insensitive mutant strain of Amycolatopsis mediterranei S699. Machine-learning approaches such as Genetic algorithm (GA), Neighborhood...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20056
更新日期：2004-04-20 00:00:00
abstract：:Although benzene can be biodegraded when dissolved oxygen is sufficient, delivering oxygen is energy intensive and can lead to air stripping the benzene. Anaerobes can biodegrade benzene by using electron acceptors other than O2 , and this may reduce costs and exposure risks; the drawback is a remarkably slower growth...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.26712
更新日期：2018-08-01 00:00:00
abstract：:In this paper an extensive batch experiment of endogenous process behavior in an aerobic biodegradation process is presented. From these experimental data, comprising measurements of MLVSS (mixed liquor volatile suspended solids) and respiration rate, in a first step the states and unknown parameters in a four-compart...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：
更新日期：2000-05-20 00:00:00
abstract：:An innovative type of biofilm model is derived by combining an individual description of microbial particles with a continuum representation of the biofilm matrix. This hybrid model retains the advantages of each approach, while providing a more realistic description of the temporal development of biofilm structure in...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20917
更新日期：2006-08-05 00:00:00
abstract：:The influence of biofilm areal cell density, species composition, and the presence of abiotic particles on the disinfection and removal of bacterial biofilms by monochloramine was investigated. Mono- and binary population biofilms of Pseudomonas aeruginosa and Klebsiella pneumoniae were grown on stainless-steel slides...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260460608
更新日期：1995-06-20 00:00:00
abstract：:Understanding of the fundamental mechanisms that govern unintentional differentiation of human induced pluripotent stem cells (hiPSCs) provides key strategies to maintain their undifferentiated state during cell expansion. This study focused on deviation from the undifferentiated state in hiPSC colonies during culture...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.25188
更新日期：2014-06-01 00:00:00
abstract：:We propose an in vitro selection strategy to identify bacteriophage variants that recognize metal ions in solution. In 6 M urea, phage T7 loses 99.9% of its activity in less than 5 min. Inactivation is accelerated by gold, but slowed by zinc and magnesium. Selection of phage over five generations in the presence of go...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260450512
更新日期：1995-03-05 00:00:00
abstract：:Animal cell cultivation in vitro has been studied for more than 40 years. However, the culture medium composition has not been designed on the basis of the stoichiometric nutritional demands for animal cell growth. In this article, a model was developed to study the stoichiometric demands for nutrients (including gluc...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260431122
更新日期：1994-05-01 00:00:00
abstract：:Whole cell pyruvate decarboxylase (PDC) from Candida utilis enhanced the enzymatic production of (R)-phenylacetylcarbinol (PAC) in an aqueous/octanol biotransformation compared to the partially purified PDC especially for a lower range of initial activities (0.3-2.5 U/mL). With an initial activity of 1.1 U/mL and at a...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20959
更新日期：2006-08-20 00:00:00
abstract：:Squalene is a lipophilic and non-volatile triterpene with many industrial applications for food, pharmaceuticals, and cosmetics. Metabolic engineering focused on optimization of the production pathway suffer from little success in improving titers because of a limited space of the cell membrane accommodating the lipop...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.27511
更新日期：2020-07-21 00:00:00
abstract：:The kinetics of cellular reproduction and the rate and extent of synthesis of extracellular polymeric substances (EPS) were investigated for P. aeruginosa growing in glucose-limited chemostats. mu(max) and K(s) estimates of 0.4 h(-1) and 2 mg glucose C/L, respectively, at 25 degrees C were obtained for this bacterium....
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260261203
更新日期：1984-12-01 00:00:00
abstract：:To build complex genetic networks with predictable behaviors, synthetic biologists use libraries of modular parts that can be characterized in isolation and assembled together to create programmable higher-order functions. Characterization experiments and computational models for gene regulatory parts operating in iso...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.26918
更新日期：2019-05-01 00:00:00
abstract：:The effect of organic solvents on the equilibrium position of lipase-catalyzed esterification of glycerol and decanoic acid has been investigated. The reaction is carried out in an aqueous-organic two-phase system. In polar solvents, high mole fractions of monoacylglycerol and low mole fractions of triacylglycerol and...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.260410113
更新日期：1993-01-05 00:00:00
abstract：:C-terminal lysine (C-K) variants are commonly observed in therapeutic monoclonal antibodies and recombinant proteins. Heterogeneity of C-K residues is believed to result from varying degree of proteolysis by endogenous carboxypeptidase(s) during cell culture production. The achievement of batch-to-batch culture perfor...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.24510
更新日期：2012-09-01 00:00:00
abstract：:A complete, integrated process for the production of an innovative formulation of penicillin G acylase from Providencia rettgeri(rPAC(P.rett))of industrial applicability is reported. In order to improve the yield of rPAC, the clone LN5.5, carrying four copies of pac gene integrated into the genome of Pichia pastoris, ...
journal_title：Biotechnology and bioengineering
pub_type： 杂志文章
doi：10.1002/bit.20728
更新日期：2006-02-05 00:00:00
