Biomedical subjects

R M Worden

Publications and source records attributed to R M Worden.

7 recordsLinked to original sources

PubMed2000

Enhancement of mass transfer using colloidal liquid aphrons: measurement of mass transfer coefficients in liquid-liquid extraction.

Interphase mass transfer of a sparingly soluble solute is often the rate-limiting step in multiphase biocatalytic processes. Colloidal liquid aphrons (CLA) provide very large interfacial areas, and thus could enhance mass transfer in such processes. The aim of this study was to characterize mass transfer properties of CLA dispersions during transfer of heptanoic acid from water to limonene. The interfacial area per unit volume (a), film mass transfer coefficient (K(L)), and volumetric mass transfer coefficient (K(L)a) values were determined in a stirred-tank reactor. These results were used, along with a literature correlation, to estimate the mass transfer coefficient of the surfactant-stabilized shell surrounding the CLA. The very large increase in a provided by the CLA was only partially offset by a slight increase in the mass transfer resistance of the shell. As a result, the range of K(L)a values obtained using CLA was about an order of magnitude greater than that obtained using a conventional dispersion. The concentration of the aqueous-phase surfactant used to form the CLA strongly affected the Sauter mean diameter of the CLA; however, the concentration of the nonpolar-phase surfactant had little effect. These results suggest that CLA have considerable potential for multiphase biocatalytic applications.

Biotechnology↗

PubMed1999

Fed-batch fermentor synthesis of 3-dehydroshikimic acid using recombinant Escherichia coli.

3-Dehydroshikimic acid (DHS), in addition to being a potent antioxidant, is the key hydroaromatic intermediate in the biocatalytic conversion of glucose into aromatic bioproducts and a variety of industrial chemicals. Microbial synthesis of DHS, like other intermediates in the common pathway of aromatic amino acid biosynthesis, has previously been examined only under shake flask conditions. In this account, synthesis of DHS using recombinant Escherichia coli constructs is examined in a fed-batch fermentor where glucose availability, oxygenation levels, and solution pH are controlled. DHS yields and titers are also determined by the activity of 3-deoxy-D-arabino-heptulosonic acid 7-phosphate (DAHP) synthase. This enzyme's expression levels, sensitivity to feedback inhibition, and the availability of its substrates, phosphoenolpyruvate (PEP) and D-erythrose 4-phosphate (E4P), dictate its in vivo activity. By combining fed-batch fermentor control with amplified expression of a feedback-insensitive isozyme of DAHP synthase and amplified expression of transketolase, DHS titers of 69 g/L were synthesized in 30% yield (mol/mol) from D-glucose. Significant concentrations of 3-dehydroquinic acid (6.8 g/L) and gallic acid (6.6 g/L) were synthesized in addition to DHS. The pronounced impact of transketolase overexpression, which increases E4P availability, on DHS titers and yields indicates that PEP availability is not a limiting factor under the fed-batch fermentor conditions employed.

3-Deoxy-7-Phosphoheptulonate Synthase↗

PubMed1998

In situ mapping of community-level cellular response with catalytic microbiosensors.

Chemotaxis, the migration of cells in the direction of a spatial chemical gradient, is important in disease progression, microbial ecology, and bioremediation. The ability to map chemoattractant gradients and the corresponding cellular growth and motility patterns is essential to the study of chemotaxis. Microelectrodes and microbiosensors have the potential to measure chemoattractant gradients with high spatial resolution. In this study, Clark-type amperometric microelectrodes and microbiosensors were used to measure solute concentrations gradients generated by a chemotactic band of Escherichia coli in a semi-solid gel. A computerized image analysis system was used to simultaneously measure the cellular concentration profile across the chemotactic band. The experimental results compared favorably with a mathematical model of solute and cell transport in the gel. Scanning electron micrographs (SEM) of micro(bio)sensor tips taken after 6 months of use showed evidence of degradation, including adhesion of foreign particles to the glass body, the adhesion of a small gel capsule to the sensor tip, and separation of the bio-interface from the tip. A needle-type microbiosensor was constructed to better protect the tip and hence increase the ruggedness of the microbiosensors.

Biosensing Techniques↗

PubMed1998

In situ mutagenesis and chemotactic selection of microorganisms in a diffusion gradient chamber.

A new method has been developed to rapidly generate and select microbial strains having increased resistance to an inhibitory compound. The method combines in situ mutagenesis with use of a continuous gradient of the inhibitor to sort cells according to their resistance levels. Microbial chemotaxis is induced to accelerate the selection process. The method was used to develop a strain of E. coli having a feedback-resistant DAHP synthase enzyme. An unsteady-state mathematical model of the process has been developed. The model, that can reproduce key trends observed experimentally, was used to explore the effects of chemotaxis on the efficiency of the selection process.

3-Deoxy-7-Phosphoheptulonate Synthase↗

PubMed1994

A diffusion gradient chamber for studying microbial behavior and separating microorganisms.

The natural habitats of most microbes are dynamic and include spatial gradients of growth substrates, electron acceptors, pH, salts, and inhibitory compounds. To mimic this diffusive aspect of nature, we developed an analytical diffusion gradient chamber (DGC) that can be used to separate, enrich for, isolate, and study the behavior of microorganisms. The chamber is a polycarbonate box containing an arena (5 by 5 by 2 cm) into which is cast a semisolid growth medium. Continuously replenished solute reservoirs positioned on each side of the arena but separated from it by a porous membrane enable the formation throughout the gel of multiple, intersecting gradients of solutes in two dimensions. With glucose as the solute, a gradient which spanned a 100-fold range in concentration was established across the arena in about 4 days. The shape of the glucose gradient was accurately predicted by a mathematical model based on Fickian diffusion. The growth and migratory behavior of Escherichia coli in response to imposed gradients of attractants (aspartate, alpha-methyl aspartate, and serine) and a repellent (valine) were examined. Cells responded in predictable ways to such gradients by forming distinctive growth and migration patterns in the DGC. This was true for wild-type E. coli as well as specific chemotaxis and motility mutants. The patterns yielded information about the threshold concentration of chemoeffectors needed to elicit a response as well as their saturating concentration. It was also evident that the metabolism of attractants significantly affected the gradients and, hence, the movement of cells. Finally, it was possible to separate E. coli and Pseudomonas fluorescens in the DGC on the basis of their differential responses to gradients of various chemoeffectors.

Journal Article↗

PubMed1991

Growth kinetics of Bacillus stearothermophilus BR219.

Bacillus stearothermophilus BR219, a phenol-resistant thermophile, can convert phenol to the specialty chemical catechol. The growth kinetics of this organism were studied in batch, continuous, and immobilized-cell culture. Batch growth was insensitive to pH between 6.0 and 8.0, but little growth occurred at 5.5. In continuous culture on a dilute medium supplemented with 10 mM phenol, several steady states were achieved between dilution rates of 0.25 and 1.3 h-1. Phenol degradation was found to be uncoupled from growth. Immobilized cells grew rapidly in a rich medium, but cell viability plummeted following a switch to a dilute medium supplemented with 5 mM phenol.

Bacteriological Techniques↗

PubMedDate not supplied

Mass-transfer properties of microbubbles. 1. Experimental studies.

Synthesis-gas fermentations have typically been gas-to-liquid mass-transfer-limited due to low solubilities of the gaseous substrates. A potential method to enhance mass-transfer rates is to sparge with microbubble dispersions. Mass-transfer coefficients for microbubble dispersions were measured in a bubble column. Oxygen microbubbles were formed in a dilute Tween 20 solution using a spinning disk apparatus. Axial dispersion coefficients measured for the bubble column ranged from 1.5 to 7.2 cm2/s and were essentially independent of flow rate. A laser-diffraction technique was used to determine the interfacial area per unit gas volume, a. The mass-transfer coefficient, KL, was determined by fitting a plug-flow model to the experimental, steady-state, liquid-phase oxygen-concentration profile. The KL values ranged from 2.9 x 10(-5) to 2.2 x 10(-4) m/s. Volumetric mass-transfer coefficients, KLa, for microbubbles with an average initial diameter of 60 microns ranged from 200 to 1800 h-1. Enhancement of mass transfer using microbubbles was demonstrated for a synthesis-gas fermentation. Butyribacterium methylotrophicum was grown in a continuous, stirred-tank reactor using a tangential filter for total cell recycle. The fermentation KLa values were 14 h-1 for conventional gas sparging through a stainless steel frit and 91 h-1 for microbubble sparging. The Power number of the microbubble generator was determined to be 0.036. Using this value, an incremental power-to-volume ratio to produce microbubbles for a B. methylotrophicum fermentation was estimated to be 0.01 kW/m3 of fermentation capacity.

Carbon Monoxide↗