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Full-Text Articles in Biomedical Engineering and Bioengineering

Analyzing Drug Release Kinetics From Water-Soluble Polymers, Sean M. Kelly, Arun K. Upadhyay, Akash Mitra, Balaji Narasimhan Feb 2019

Analyzing Drug Release Kinetics From Water-Soluble Polymers, Sean M. Kelly, Arun K. Upadhyay, Akash Mitra, Balaji Narasimhan

Chemical and Biological Engineering Publications

The ability to develop predictive mathematical models of therapeutic release from pharmaceutical formulations has enormous potential to enhance our understanding of such systems and improve the controlled release of the payload. The current work describes the development and testing of a one-dimensional model of drug transport from amorphous, swelling/dissolving polymers. Model parameters such as the diffusivities of water and drug, the initial loading of the drug, the polymer dissolution rate, drug-polymer interactions, and the tablet thickness were varied, demonstrating the ability to tune the release to be controlled by either drug diffusion or polymer chain disentanglement. In addition, predictions ...


Interleukin-1 Alpha Increases Anti-Tumor Efficacy Of Cetuximab In Head And Neck Squamous Cell Carcinoma, Madelyn Espinosa-Cotton, Samuel N. Rodman Iii, Kathleen A. Ross, Isaac J. Jensen, Kenley Sangodeyi-Miller, Ayana J. Mclaren, Rachel A. Dahl, Katherine N. Gibson-Corley, Adam T. Koch, Yang-Xin Fu, Vladimir P. Badovinac, Douglas Laux, Balaji Narasimhan, Andrean L. Simons Jan 2019

Interleukin-1 Alpha Increases Anti-Tumor Efficacy Of Cetuximab In Head And Neck Squamous Cell Carcinoma, Madelyn Espinosa-Cotton, Samuel N. Rodman Iii, Kathleen A. Ross, Isaac J. Jensen, Kenley Sangodeyi-Miller, Ayana J. Mclaren, Rachel A. Dahl, Katherine N. Gibson-Corley, Adam T. Koch, Yang-Xin Fu, Vladimir P. Badovinac, Douglas Laux, Balaji Narasimhan, Andrean L. Simons

Chemical and Biological Engineering Publications

Background: Despite the high prevalence of epidermal growth factor receptor (EGFR) overexpression in head and neck squamous cell carcinomas (HNSCCs), incorporation of the EGFR inhibitor cetuximab into the clinical management of HNSCC has not led to significant changes in long-term survival outcomes. Therefore, the identification of novel therapeutic approaches to enhance the clinical efficacy of cetuximab could lead to improved long-term survival for HNSCC patients. Our previous work suggests that EGFR inhibition activates the interleukin-1 (IL-1) pathway via tumor release of IL-1 alpha (IL-1α), although the clinical implications of activating this pathway are unclear in the context of cetuximab therapy ...


Treatment Of Neurodegenerative Disorders Through The Blood-Brain Barrier Using Nanocarriers, Nitya Poovaiah, Zahra Davoudi, Haisheng Peng, Benjamin Schlichtmann, Surya K. Mallapragada, Balaji Narasimhan, Qun Wang Aug 2018

Treatment Of Neurodegenerative Disorders Through The Blood-Brain Barrier Using Nanocarriers, Nitya Poovaiah, Zahra Davoudi, Haisheng Peng, Benjamin Schlichtmann, Surya K. Mallapragada, Balaji Narasimhan, Qun Wang

Chemical and Biological Engineering Publications

Neurodegenerative diseases refer to disorders of the central nervous system (CNS) that are caused by neuronal degradations, dysfunctions, or death. Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease (APHD) are regarded as the three major neurodegenerative diseases. There is a vast body of literature on the causes and treatments of these neurodegenerative diseases. However, the main obstacle in developing an effective treatment strategy is the permeability of the treatment components to the blood-brain barrier (BBB). Several strategies have been developed to improve this obstruction. For example, nanomaterials facilitate drug delivery to the BBB due to their size. They ...


Escherichia Coli Attachment To Model Particulates: The Effects Of Bacterial Cell Characteristics And Particulate Properties, Xiao Liang, Chunyu Liao, Michelle L. Soupir, Laura R. Jarboe, Michael L. Thompson, Philip M. Dixon Sep 2017

Escherichia Coli Attachment To Model Particulates: The Effects Of Bacterial Cell Characteristics And Particulate Properties, Xiao Liang, Chunyu Liao, Michelle L. Soupir, Laura R. Jarboe, Michael L. Thompson, Philip M. Dixon

Chemical and Biological Engineering Publications

E. coli bacteria move in streams freely in a planktonic state or attached to suspended particulates. Attachment is a dynamic process, and the fraction of attached microorganisms is thought to be affected by both bacterial characteristics and particulate properties. In this study, we investigated how the properties of cell surfaces and stream particulates influence attachment. Attachment assays were conducted for 77 E. coli strains and three model particulates (ferrihydrite, Ca-montmorillonite, or corn stover) under environmentally relevant conditions. Surface area, particle size distribution, and total carbon content were determined for each type of particulate. Among the three particulates, attachment fractions to ...


Anticancer Drugs, Le Zhao, Zengyi Shao, Jacqueline V. Shanks Jan 2017

Anticancer Drugs, Le Zhao, Zengyi Shao, Jacqueline V. Shanks

Chemical and Biological Engineering Publications

Plant‐derived anticancer drugs play a large role in anticancer pharmaceuticals. Through reviewing the four major types of plant anticancer drugs, namely vinca alkaloids, taxane diterpenoids, podophyllotoxin lignans, and camptothecin quinoline alkaloids, this article illustrates the development process, current status, existing challenges, and future perspective of the plant anticancer drug production. Moreover, this review explains how various biotechnologies, from the mature elicitation strategy to the “omics” techniques that are still undergoing development, can be applied to address the challenges in improving the production of the plant‐sourced anticancer drugs.


High Throughput Studies Of Cell Migration In 3d Microtissues Fabricated By A Droplet Microfluidic Chip, Xiangchen Che, Jacob A. M. Nuhn, Ian C. Schneider, Long Que Jan 2016

High Throughput Studies Of Cell Migration In 3d Microtissues Fabricated By A Droplet Microfluidic Chip, Xiangchen Che, Jacob A. M. Nuhn, Ian C. Schneider, Long Que

Chemical and Biological Engineering Publications

Arrayed three-dimensional (3D) micro-sized tissues with encapsulated cells (microtissues) have been fabricated by a droplet microfluidic chip. The extracellular matrix (ECM) is a polymerized collagen network. One or multiple breast cancer cells were embedded within the microtissues, which were stored in arrayed microchambers on the same chip without ECM droplet shrinkage over 48 h. The migration trajectory of the cells was recorded by optical microscopy. The migration speed was calculated in the range of 3–6 µm/h. Interestingly, cells in devices filled with a continuous collagen network migrated faster than those where only droplets were arrayed in the chambers ...


Responsive Pentablock Copolymers For Sirna Delivery, Surya K. Mallapragada, Metin Uz, Sacide Alsoy Altinkaya Jan 2015

Responsive Pentablock Copolymers For Sirna Delivery, Surya K. Mallapragada, Metin Uz, Sacide Alsoy Altinkaya

Chemical and Biological Engineering Publications

In this study, temperature and pH responsive cationic and amphiphilic pentablock copolymers, which consist of the temperature responsive triblock Pluronic F127 sandwiched between pH responsive PDEAEM (poly(2-diethylaminoethyl methacrylate)) end blocks, were used for the first time in the development of polyplex and gold nanoparticle (AuNP) based multicomponent siRNA delivery systems (MCSs). Copolymers in both systems protected siRNA from external effects, provided cell entry and endosomal escape. The thermoreversible micellization of the hydrophobic PPO block facilitated the cellular entry while the PDEAEM blocks enhanced the endosomal escape through protonated tertiary amine groups by pH buffering. The synergistic advantages of the ...


Understanding Biocatalyst Inhibition By Carboxylic Acids, Laura R. Jarboe, Liam Royce, Ping Liu Sep 2013

Understanding Biocatalyst Inhibition By Carboxylic Acids, Laura R. Jarboe, Liam Royce, Ping Liu

Chemical and Biological Engineering Publications

Carboxylic acids are an attractive biorenewable chemical in terms of their flexibility and usage as precursors for a variety of industrial chemicals. It has been demonstrated that such carboxylic acids can be fermentatively produced using engineered microbes, such as Escherichia coli andSaccharomyces cerevisiae. However, like many other attractive biorenewable fuels and chemicals, carboxylic acids become inhibitory to these microbes at concentrations below the desired yield and titer. In fact, their potency as microbial inhibitors is highlighted by the fact that many of these carboxylic acids are routinely used as food preservatives. This review highlights the current knowledge regarding the ...


Substrate Binding By The Catalytic Domain And Carbohydrate Binding Module Of Ruminococcus Flavefaciens Fd-1 Xyloglucanase/ Endoglucanase, Christopher D. Warner, Gulden Camci-Unal, Nicola L.B. Pohl, Clark Ford, Peter J. Reilly Jan 2013

Substrate Binding By The Catalytic Domain And Carbohydrate Binding Module Of Ruminococcus Flavefaciens Fd-1 Xyloglucanase/ Endoglucanase, Christopher D. Warner, Gulden Camci-Unal, Nicola L.B. Pohl, Clark Ford, Peter J. Reilly

Chemical and Biological Engineering Publications

Binding and thermodynamic properties of a carbohydrate binding module (CBM) and a glycoside hydrolase family 44 xyloglucanase/endoglucanase catalytic domain (CD) fromRuminococcus flavefaciens, both when separate and when linked to each other, have been quantified when binding various β-1,4-linked glucans and xylans. The three constructs bind cellotetraose, cellopentaose, and cellohexaose with association constants that increase with chain length. The CBM does not bind xylotetraose, xylopentaose, or xylohexaose. The CD appears to bind carboxymethylcellulose (CMC) and xylan only weakly, while the CBM and the CD/CBM bind them much more strongly than they bind the cellooligosaccharides. CMC is bound ...


Integrated Self-Assembly Of The Mms6 Magnetosome Protein To Form An Iron-Responsive Structure, Shuren Feng, Lijun Wang, Pierre Palo, Xunpei Liu, Surya K. Mallapragada, Marit Nilsen-Hamilton Jan 2013

Integrated Self-Assembly Of The Mms6 Magnetosome Protein To Form An Iron-Responsive Structure, Shuren Feng, Lijun Wang, Pierre Palo, Xunpei Liu, Surya K. Mallapragada, Marit Nilsen-Hamilton

Chemical and Biological Engineering Publications

A common feature of biomineralization proteins is their self-assembly to produce a surface consistent in size with the inorganic crystals that they produce. Mms6, a small protein of 60 amino acids from Magnetospirillum magneticum strain AMB-1 that promotes the in vitro growth of superparamagnetic magnetite nanocrystals, assembles in aqueous solution to form spherical micelles that could be visualized by TEM and AFM. The results reported here are consistent with the view that the N and C-terminal domains interact with each other within one polypeptide chain and across protein units in the assembly. From studies to determine the amino acid residues ...


Growth Condition Optimization For Docosahexaenoic Acid (Dha) Production By Moritella Marina Mp-1, Kumar Babu Kautharapu, John A. Rathmacher, Laura R. Jarboe Jan 2013

Growth Condition Optimization For Docosahexaenoic Acid (Dha) Production By Moritella Marina Mp-1, Kumar Babu Kautharapu, John A. Rathmacher, Laura R. Jarboe

Chemical and Biological Engineering Publications

The marine organism Moritella marina MP-1 produces the polyunsaturated fatty acid docosahexaenoic acid (DHA). While the basic metabolic pathway for DHA production in this organism has been identified, the impact of growth conditions on DHA production is largely unknown. This study examines the effect of supplemental carbon, nitrogen and salts, growth temperature and media composition and pH on DHA and biomass production and the fatty acid profile. The addition of supplemental nitrogen significantly increased the overall DHA titer via an increase in biomass production. Supplemental glucose or glycerol increased biomass production, but decreased the amount of DHA per biomass, resulting ...


Optimization Of Enzyme Parameters For Fermentative Production Of Biorenewable Fuels And Chemicals, Laura R. Jarboe, Ping Liu, Kumar Babu Kautharapu, Lonnie O. Ingram Oct 2012

Optimization Of Enzyme Parameters For Fermentative Production Of Biorenewable Fuels And Chemicals, Laura R. Jarboe, Ping Liu, Kumar Babu Kautharapu, Lonnie O. Ingram

Chemical and Biological Engineering Publications

Microbial biocatalysts such as Escherichia coli and Saccharomyces cerevisiae have been extensively subjected to Metabolic Engineering for the fermentative production of biorenewable fuels and chemicals. This often entails the introduction of new enzymes, deletion of unwanted enzymes and efforts to fine-tune enzyme abundance in order to attain the desired strain performance. Enzyme performance can be quantitatively described in terms of the Michaelis-Menten type parameters Km, turnover number kcat and Ki, which roughly describe the affinity of an enzyme for its substrate, the speed of a reaction and the enzyme sensitivity to inhibition by regulatory molecules. Here we describe examples of ...


Metabolic Engineering Of Biocatalysts For Carboxylic Acids Production, Ping Liu, Laura R. Jarboe Oct 2012

Metabolic Engineering Of Biocatalysts For Carboxylic Acids Production, Ping Liu, Laura R. Jarboe

Chemical and Biological Engineering Publications

Fermentation of renewable feedstocks by microbes to produce sustainable fuels and chemicals has the potential to replace petrochemical-based production. For example, carboxylic acids produced by microbial fermentation can be used to generate primary building blocks of industrial chemicals by either enzymatic or chemical catalysis. In order to achieve the titer, yield and productivity values required for economically viable processes, the carboxylic acid-producing microbes need to be robust and wellperforming. Traditional strain development methods based on mutagenesis have proven useful in the selection of desirable microbial behavior, such as robustness and carboxylic acid production. On the other hand, rationally-based metabolic engineering ...


Structural Classification Of Biotin Carboxyl Carrier Proteins, Yingfei Chen, Armando Elizondo-Noriega, David C. Cantu, Peter J. Reilly Oct 2012

Structural Classification Of Biotin Carboxyl Carrier Proteins, Yingfei Chen, Armando Elizondo-Noriega, David C. Cantu, Peter J. Reilly

Chemical and Biological Engineering Publications

We gathered primary and tertiary structures of acyl-CoA carboxylases from public databases, and established that members of their biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) domains occur in one family each and that members of their carboxyl transferase (CT) domains occur in two families. Protein families have members similar in primary and tertiary structure that probably have descended from the same protein ancestor. The BCCP domains complexed with biotin in acyl and acyl-CoA carboxylases transfer bicarbonate ions from BC domains to CT domains, enabling the latter to carboxylate acyl and acyl-CoA moieties. We separated the BCCP domains into ...


Docking Studies On Glycoside Hydrolase Family 47 Endoplasmic Reticulum Α-(1→2)-Mannosidase I To Elucidate The Pathway To The Substrate Transition State, Chandrika Mulakala, Wim Nerinckx, Peter J. Reilly Sep 2012

Docking Studies On Glycoside Hydrolase Family 47 Endoplasmic Reticulum Α-(1→2)-Mannosidase I To Elucidate The Pathway To The Substrate Transition State, Chandrika Mulakala, Wim Nerinckx, Peter J. Reilly

Chemical and Biological Engineering Publications

α-(1→2)-Mannosidase I from the endoplasmic reticulum (ERManI), a Family 47 glycoside hydrolase, is a key enzyme in the N-glycan synthesis pathway. Catalytic-domain crystal structures of yeast and human ERMan1s have been determined, the former with a hydrolytic product and the latter without ligands, with the inhibitors 1-deoxymannojirimycin and kifunensine, and with a thiodisaccharide substrate analog. Both inhibitors were bound at the base of the funnel-shaped active site as the unusual 1C4 conformer, while the substrate analog glycon is a 3S1 conformer. In the current study, AutoDock was used to dock α-d-mannopyranosyl-(1 ...


Block Copolymer-Quantum Dot Micelles For Multienzyme Colocalization, Feng Jia, Yanjie Zhang, Balaji Narasimhan, Surya K. Mallapragada Jan 2012

Block Copolymer-Quantum Dot Micelles For Multienzyme Colocalization, Feng Jia, Yanjie Zhang, Balaji Narasimhan, Surya K. Mallapragada

Chemical and Biological Engineering Publications

To mimic the structure and functionality of multienzyme complexes, which are widely present in Nature, Pluronic-based micelles were designed to colocalize multiple enzymes. To stabilize the micelles as well as to enable characterization of single enzyme immobilization and multienzyme colocalization by Förster resonance energy transfer (FRET), quantum dots (QDs) were incorporated into the micelles to form Pluronic-QD micelles using a novel microreactor. Model enzymes glucose oxidase (GOX) and horseradish peroxidase (HRP) were respectively labeled with fluorescent dyes. The results indicated that FRET occurred between the QDs and dyes that labeled each type of enzyme in single enzyme immobilization studies ...


Phylogenetic And Experimental Characterization Of An Acyl-Acp Thioesterase Family Reveals Significant Diversity In Enzymatic Specificity And Activity, Fuyuan Jing, David C. Cantu, Jarmila Tvaruzkova, Jay P. Chipman, Basil J. Nikolau, Marna D. Yandeau-Nelson, Peter J. Reilly Aug 2011

Phylogenetic And Experimental Characterization Of An Acyl-Acp Thioesterase Family Reveals Significant Diversity In Enzymatic Specificity And Activity, Fuyuan Jing, David C. Cantu, Jarmila Tvaruzkova, Jay P. Chipman, Basil J. Nikolau, Marna D. Yandeau-Nelson, Peter J. Reilly

Chemical and Biological Engineering Publications

Background: Acyl-acyl carrier protein thioesterases (acyl-ACP TEs) catalyze the hydrolysis of the thioester bond that links the acyl chain to the sulfhydryl group of the phosphopantetheine prosthetic group of ACP. This reaction terminates acyl chain elongation of fatty acid biosynthesis, and in plant seeds it is the biochemical determinant of the fatty acid compositions of storage lipids.

Results: To explore acyl-ACP TE diversity and to identify novel acyl ACP-TEs, 31 acyl-ACP TEs from wide-ranging phylogenetic sources were characterized to ascertain their in vivo activities and substrate specificities. These acylACP TEs were chosen by two different approaches: 1) 24 TEs were ...


Kinetic Characterization Of A Glycoside Hydrolase Family 44 Xyloglucanase/Endoglucanase From Ruminococcus Flavefaciens Fd-1, Christopher D. Warner, Rica M. Go, Carolina García-Salinas, Clark Ford, Peter J. Reilly Jan 2011

Kinetic Characterization Of A Glycoside Hydrolase Family 44 Xyloglucanase/Endoglucanase From Ruminococcus Flavefaciens Fd-1, Christopher D. Warner, Rica M. Go, Carolina García-Salinas, Clark Ford, Peter J. Reilly

Chemical and Biological Engineering Publications

Two forms of Ruminococcus flavefaciens FD-1 endoglucanase B, a member of glycoside hydrolase family 44, one with only a catalytic domain and the other with a catalytic domain and a carbohydrate binding domain (CBM), were produced. Both forms hydrolyzed cellotetraose, cellopentaose, cellohexaose, carboxymethylcellulose (CMC), birchwood and larchwood xylan, xyloglucan, lichenan, and Avicel but not cellobiose, cellotriose, mannan, or pullulan. Addition of the CBM increased catalytic efficiencies on both CMC and birchwood xylan but not on xyloglucan, and it decreased rates of cellopentaose and cellohexaose hydrolysis. Catalytic efficiencies were much higher on xyloglucan than on other polysaccharides. Hydrolysis rates increased with ...


Engineering Ethanologenic Escherichia Coli For Levoglucosan Utilization, Donovan S. Layton, Avanthi Ajjarapu, Dong Won Choi, Laura R. Jarboe Jan 2011

Engineering Ethanologenic Escherichia Coli For Levoglucosan Utilization, Donovan S. Layton, Avanthi Ajjarapu, Dong Won Choi, Laura R. Jarboe

Chemical and Biological Engineering Publications

Levoglucosan is a major product of biomass pyrolysis. While this pyrolyzed biomass, also known as bio-oil, contains sugars that are an attractive fermentation substrate, commonly-used biocatalysts, such as Escherichia coli, lack the ability to metabolize this anhydrosugar. It has previously been shown that recombinant expression of the levoglucosan kinase enzyme enables use of levoglucosan as carbon and energy source. Here, ethanologenic E. coli KO11 was engineered for levoglucosan utilization by recombinant expression of levoglucosan kinase from Lipomyces starkeyi. Our engineering strategy uses a codon-optimized gene that has been chromosomally integrated within the pyruvate to ethanol (PET) operon and does not ...


Thyme: A Database For Thioester-Active Enzymes, David C. Cantu, Yingfei Chen, Matthew L. Lemons, Peter J. Reilly Nov 2010

Thyme: A Database For Thioester-Active Enzymes, David C. Cantu, Yingfei Chen, Matthew L. Lemons, Peter J. Reilly

Chemical and Biological Engineering Publications

The ThYme (Thioester-active enzYme;http://www.enzyme.cbirc.iastate.edu) database has been constructed to bring together amino acid sequences and 3D (tertiary) structures of all the enzymes constituting the fatty acid synthesis and polyketide synthesis cycles. These enzymes are active on thioester-containing substrates, specifically those that are parts of the acyl-CoA synthase, acyl-CoA carboxylase, acyl transferase, ketoacyl synthase, ketoacyl reductase, hydroxyacyl dehydratase, enoyl reductase and thioesterase enzyme groups. These groups have been classified into families, members of which are similar in sequences, tertiary structures and catalytic mechanisms, implying common protein ancestry. ThYme is continually updated as sequences and tertiary ...


Mechanism Of Xylobiose Hydrolysis By Gh43 Β-Xylosidase, Ian J. Barker, Luis Petersen, Peter J. Reilly Oct 2010

Mechanism Of Xylobiose Hydrolysis By Gh43 Β-Xylosidase, Ian J. Barker, Luis Petersen, Peter J. Reilly

Chemical and Biological Engineering Publications

Glycoside hydrolases cleave the glycosidic linkage between two carbohydrate moieties. They are among the most efficient enzymes currently known. β-Xylosidases from glycoside hydrolase family 43 hydrolyze the nonreducing ends of xylooligomers using an inverting mechanism. Although the general mechanism and catalytic amino acid residues of β-xylosidases are known, the nature of the reaction’s transition state and the conformations adopted by the glycon xylopyranosyl ring along the reaction pathway are still elusive. In this work, the xylobiose hydrolysis reaction catalyzed by XynB3, a β-xylosidase produced byGeobacillus stearothermophilus T-6, was explicitly modeled using first-principles quantum mechanics/molecular mechanics Car−Parrinello ...


Molecular Mechanism Of The Glycosylation Step Catalyzed By Golgi Α-Mannosidase Ii: A Qm/Mm Metadynamics Investigation, Luis Petersen, Albert Ardèvol, Carme Rovira, Peter J. Reilly May 2010

Molecular Mechanism Of The Glycosylation Step Catalyzed By Golgi Α-Mannosidase Ii: A Qm/Mm Metadynamics Investigation, Luis Petersen, Albert Ardèvol, Carme Rovira, Peter J. Reilly

Chemical and Biological Engineering Publications

Golgi α-mannosidase II (GMII), a member of glycoside hydrolase family 38, cleaves two mannosyl residues from GlcNAcMan5GlcNAc2 as part of the N-linked glycosylation pathway. To elucidate the molecular and electronic details of the reaction mechanism, in particular the conformation of the substrate at the transition state, we performed quantum mechanics/molecular mechanics metadynamics simulations of the glycosylation reaction catalyzed by GMII. The calculated free energy of activation for mannosyl glycosylation (23 kcal/mol) agrees very well with experiments, as does the conformation of the glycon mannosyl ring in the product of the glycosylation reaction (the covalent intermediate ...


Sensing Polymer/Dna Polyplex Dissociation Using Quantum Dot Fluorophores, Bingqi Zhang, Yanjie Zhang, Surya K. Mallapragada, Aaron R. Clapp Jan 2010

Sensing Polymer/Dna Polyplex Dissociation Using Quantum Dot Fluorophores, Bingqi Zhang, Yanjie Zhang, Surya K. Mallapragada, Aaron R. Clapp

Chemical and Biological Engineering Publications

We characterized the dissociation of polymer/DNA polyplexes designed for gene delivery using water-soluble quantum dots (QDs). A pH-responsive pentablock copolymer was designed to form stable complexes with plasmid DNA via tertiary amine segments. Dissociation of the polyplex was induced using chloroquine where the efficiency of this process was sensed through changes in QD fluorescence. We found that increasing concentrations of pentablock copolymer and DNA led to quenching of QD fluorescence, while chloroquine alone had no measurable effect. The mechanism of quenching was elucidated by modeling the process as the combination of static and dynamic quenching from the pentablock copolymer ...


Utilizing Protein-Lean Coproducts From Corn Containing Recombinant Pharmaceutical Proteins For Ethanol Production, Ilankovan Paraman, Lorena Beatriz Moeller, M. Paul Scott, Kan Wang, Charles E. Glatz, Lawrence A. Johnson Jan 2010

Utilizing Protein-Lean Coproducts From Corn Containing Recombinant Pharmaceutical Proteins For Ethanol Production, Ilankovan Paraman, Lorena Beatriz Moeller, M. Paul Scott, Kan Wang, Charles E. Glatz, Lawrence A. Johnson

Chemical and Biological Engineering Publications

Protein-lean fractions of corn (maize) containing recombinant (r) pharmaceutical proteins were evaluated as a potential feedstock to produce fuel ethanol. The levels of residual r-proteins in the coproduct, distillers dry grains with solubles (DDGS), were determined. Transgenic corn lines containing recombinant green fluorescence protein (r-GFP) and a recombinant subunit vaccine of Escherichia coli enterotoxin (r-LTB), primarily expressed in endosperm, and another two corn lines containing recombinant human collagen (r-CIα1) andr-GFP, primarily expressed in germ, were used as model systems. The kernels were either ground and used for fermentation or dry fractionated to recover germ-rich ...


Metabolic Analysis Of Wild-Type Escherichia Coli And A Pyruvate Dehydrogenase Complex (Pdhc)-Deficient Derivative Reveals The Role Of Pdhc In The Fermentative Metabolism Of Glucose, Abhishek Murarka, James M. Clomburg, Sean Moran, Jacqueline V. Shanks, Ramon Gonzalez Jan 2010

Metabolic Analysis Of Wild-Type Escherichia Coli And A Pyruvate Dehydrogenase Complex (Pdhc)-Deficient Derivative Reveals The Role Of Pdhc In The Fermentative Metabolism Of Glucose, Abhishek Murarka, James M. Clomburg, Sean Moran, Jacqueline V. Shanks, Ramon Gonzalez

Chemical and Biological Engineering Publications

Pyruvate is located at a metabolic junction of assimilatory and dissimilatory pathways and represents a switch point between respiratory and fermentative metabolism. In Escherichia coli, the pyruvate dehydrogenase complex (PDHC) and pyruvate formate-lyase are considered the primary routes of pyruvate conversion to acetyl-CoA for aerobic respiration and anaerobic fermentation, respectively. During glucose fermentation, the in vivo activity of PDHC has been reported as either very low or undetectable, and the role of this enzyme remains unknown. In this study, a comprehensive characterization of wild-type E. coli MG1655 and a PDHC-deficient derivative (Pdh) led to the identification of the role of ...


Tertiary Structure And Characterization Of A Glycoside Hydrolase Family 44 Endoglucanase From Clostridium Acetobutylicum, Christopher D. Warner, Julie A. Hoy, Taran C. Shilling, Michael J. Linnen, Nathaniel D. Ginder, Clark Ford, Richard B. Honzatko, Peter J. Reilly Jan 2010

Tertiary Structure And Characterization Of A Glycoside Hydrolase Family 44 Endoglucanase From Clostridium Acetobutylicum, Christopher D. Warner, Julie A. Hoy, Taran C. Shilling, Michael J. Linnen, Nathaniel D. Ginder, Clark Ford, Richard B. Honzatko, Peter J. Reilly

Chemical and Biological Engineering Publications

A gene encoding a glycoside hydrolase family 44 (GH44) protein from Clostridium acetobutylicum ATCC 824 was synthesized and transformed into Escherichia coli.The previously uncharacterized protein was expressed with a C-terminal His tag and purified by nickel-nitrilotriacetic acid affinity chromatography. Crystallization and X-ray diffraction to a 2.2-Å resolution revealed a triose phosphate isomerase (TIM) barrel-like structure with additional Greek key and β-sandwich folds, similar to other GH44 crystal structures. The enzyme hydrolyzes cellotetraose and larger cellooligosaccharides, yielding an unbalanced product distribution, including some glucose. It attacks carboxymethylcellulose and xylan at approximately the same rates. Its activity on carboxymethylcellulose is ...


Metabolic Engineering For Production Of Biorenewable Fuels And Chemicals: Contributions Of Synthetic Biology, Laura R. Jarboe, Xueli Zhang, Xuan Wang, Jonathan C. Moore, K. T. Shanmugam, Lonnie O. Ingram Jan 2010

Metabolic Engineering For Production Of Biorenewable Fuels And Chemicals: Contributions Of Synthetic Biology, Laura R. Jarboe, Xueli Zhang, Xuan Wang, Jonathan C. Moore, K. T. Shanmugam, Lonnie O. Ingram

Chemical and Biological Engineering Publications

Production of fuels and chemicals through microbial fermentation of plant material is a desirable alternative to petrochemicalbased production. Fermentative production of biorenewable fuels and chemicals requires the engineering of biocatalysts that can quickly and efficiently convert sugars to target products at a cost that is competitive with existing petrochemical-based processes. It is also important that biocatalysts be robust to extreme fermentation conditions, biomass-derived inhibitors, and their target products. Traditional metabolic engineering has made great advances in this area, but synthetic biology has contributed and will continue to contribute to this field, particularly with next-generation biofuels. This work reviews the use ...


Mechanisms Of Aqueous Extraction Of Soybean Oil, K. A. Campbell, Charles E. Glatz Nov 2009

Mechanisms Of Aqueous Extraction Of Soybean Oil, K. A. Campbell, Charles E. Glatz

Chemical and Biological Engineering Publications

Aqueous extraction processing (AEP) of soy is a promising green alternative to hexane extraction processing. To improve AEP oil yields, experiments were conducted to probe the mechanisms of oil release. Microscopy of extruded soy before and after extraction with and without protease indicated that unextracted oil is sequestered in an insoluble matrix of denatured protein and is released by proteolytic digestion of this matrix. In flour from flake, unextracted oil is contained as intact oil bodies in undisrupted cells, or as coalesced oil droplets too large to pass out of the disrupted cellular matrix. Our results suggest that emulsification is ...


Twisting Of Glycosidic Bonds By Hydrolases, Glenn P. Johnson, Luis Petersen, Alfred D. French, Peter J. Reilly Nov 2009

Twisting Of Glycosidic Bonds By Hydrolases, Glenn P. Johnson, Luis Petersen, Alfred D. French, Peter J. Reilly

Chemical and Biological Engineering Publications

Patterns of scissile bond twisting have been found in crystal structures of glycoside hydrolases (GHs) that are complexed with substrates and inhibitors. To estimate the increased potential energy in the substrates that results from this twisting, we have plotted torsion angles for the scissile bonds on hybrid Quantum Mechanics::Molecular Mechanics energy surfaces. Eight such maps were constructed, including one for α-maltose and three for different forms of methyl α-acarviosinide to provide energies for twisting of α-(1,4) glycosidic bonds. Maps were also made for β-thiocellobiose and for three β-cellobiose conformers having different glycon ring shapes to model distortions ...


Mechanism Of Cellulose Hydrolysis By Inverting Gh8 Endoglucanases: A Qm/Mm Metadynamics Study, Luis Petersen, Albert Ardèvol, Carme Rovira, Peter J. Reilly Apr 2009

Mechanism Of Cellulose Hydrolysis By Inverting Gh8 Endoglucanases: A Qm/Mm Metadynamics Study, Luis Petersen, Albert Ardèvol, Carme Rovira, Peter J. Reilly

Chemical and Biological Engineering Publications

A detailed understanding of the catalytic strategy of cellulases is key to finding alternative ways to hydrolyze cellulose to mono-, di-, and oligosaccharides. Endoglucanases from glycoside hydrolase family 8 (GH8) catalyze the hydrolysis of β-1,4-glycosidic bonds in cellulose by an inverting mechanism believed to involve a oxacarbenium ion-like transition state (TS) with a boat-type conformation of the glucosyl unit in subsite −1. In this work, hydrolysis by Clostridium thermocellum endo-1,4-glucanase A was computationally simulated with quantum mechanics/molecular mechanics metadynamics based on density functional theory. Our calculations show that the glucosyl residue in subsite −1 in the Michaelis ...