A5058270282- Biofuel production and bioconversion
- Bacterial Genetics and Biotechnology
- Bacteriophages and microbial interactions
- Enzyme Production and Characterization
- Microbial Metabolic Engineering and Bioproduction
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Microfluidic and Capillary Electrophoresis Applications
- Protein purification and stability
- Virus-based gene therapy research
- Biosimilars and Bioanalytical Methods
- Genomics and Phylogenetic Studies
- Escherichia coli research studies
- Immune cells in cancer
- Studies on Chitinases and Chitosanases
- Bioenergy crop production and management
- Enzyme-mediated dye degradation
- Fungal and yeast genetics research
- Endoplasmic Reticulum Stress and Disease
- Extracellular vesicles in disease
- Respiratory viral infections research
- Fermentation and Sensory Analysis
- Mycorrhizal Fungi and Plant Interactions
- Antibiotic Resistance in Bacteria
- RNA Interference and Gene Delivery
R&D Systems (United States)
2020-2024
ProteinSimple (United States)
2020-2023
University of California, Santa Barbara
2014-2021
Cornell University
2010-2017
Indiana University Bloomington
2014
Zero to Three
2010
The fungal kingdom is the source of almost all industrial enzymes in use for lignocellulose bioprocessing. We developed a systems-level approach that integrates transcriptomic sequencing, proteomics, phenotype, and biochemical studies relatively unexplored basal fungi. Anaerobic gut fungi isolated from herbivores produce large array biomass-degrading synergistically degrade crude, untreated plant biomass are competitive with optimized commercial preparations Aspergillus Trichoderma. Compared...
The Campylobacter jejuni pgl gene cluster encodes a complete N-linked protein glycosylation pathway that can be functionally transferred into Escherichia coli. In this system, we analyzed the interplay between glycosylation, membrane translocation and folding of acceptor proteins in bacteria. We developed recombinant N-glycan peptide tag permits diverse expressed periplasm glycosylation-competent E. coli cells. With "glycosylation tag," clear difference was observed patterns found on...
A widespread feature in the genomes of most bacteria and archaea is an array clustered, regularly interspaced short palindromic repeats (CRISPRs) that, together with a group CRISPR-associated (Cas) proteins, mediate immunity against invasive nucleic acids such as plasmids viruses. Here, CRISPR-Cas system was activated cells expressing plasmid-encoded protein that targeted to twin-arginine translocation (Tat) pathway. Expression this Tat substrate resulted upregulation Cas enzymes subsequent...
The CRISPR (clustered regularly interspaced short palindromic repeats) system is a prokaryotic RNA-based adaptive immune against extrachromosomal genetic elements. Cas2 universally conserved core CRISPR-associated protein required for the acquisition of new spacers adaptation. It was previously characterized as an endoribonuclease with preference single-stranded (ss)RNA. Here, we show using crystallography, mutagenesis, and isothermal titration calorimetry that Bacillus halodurans (Bha_Cas2)...
Cellulosomes are synthesized by anaerobic bacteria and fungi to degrade lignocellulose via synergistic action of multiple enzymes fused a protein scaffold. Through templating key domains (cohesin dockerin), designer cellulosomes have been engineered from bacterial motifs alter the activity, stability, degradation efficiency enzyme complexes. Recently parts list for fungal (Neocallimastigomycota) was determined, which revealed sequence divergent cohesin, dockerin, scaffoldin that could be...
Agrobacterium tumefaciens can adhere to plant tissues and abiotic surfaces forms biofilms. Cell surface appendages called pili play an important role in adhesion biofilm formation diverse bacterial systems. The A. C58 genome sequence revealed the presence of ctpABCDEFGHI genes (cluster type IV pili; Atu0216 Atu0224), homologous tad-type pilus systems from several bacteria, including Aggregatibacter actinomycetemcomitans Caulobacter crescentus. These fall into IVb group, which function...
Anaerobic fungi are among the most active plant‐degrading microbes in nature. Increased insight into mechanisms and environmental cues that regulate fungal hydrolysis would better inform bioprocessing strategies to depolymerize lignocellulose. Here, we compare response of three strains anaerobic ( Piromyces finnis , Anaeromyces robustus Neocallimastix californiae ) catabolite regulation by simple carbohydrates. exhibited high enzymatic activity against crystalline cellulose, which was...
A variety of strategies now exist for the extracellular expression recombinant proteins using laboratory strains Escherichia coli . However, secreted often accumulate in culture medium at levels that are too low to be practically useful most synthetic biology and metabolic engineering applications. The situation is compounded by lack generalized screening tools optimizing secretion process. To address this challenge, we developed a genetic approach studying protein-secretion pathways E....
The extracellular expression of recombinant proteins using laboratory strains Escherichia coli is now routinely achieved naturally secreted substrates, such as YebF or the osmotically inducible protein Y (OsmY), carrier molecules. However, secretion efficiency through these pathways needs to be improved for most synthetic biology and metabolic engineering applications. To address this challenge, we developed a generalizable survival-based selection strategy that effectively couples...
Anaerobic fungi (Neocallimastigomycota) isolated from the guts of herbivores are powerful biomass-degrading organisms that enhance their degradative ability through formation cellulosomes, multienzyme complexes synergistically colocalize enzymes to extract sugars recalcitrant plant matter. However, a functional understanding how fungal cellulosomes deployed in vivo orchestrate matter degradation is lacking, as knowledge cellulosome production and function vary throughout morphologically...
Post-translational modifications (PTMs) of adeno-associated virus (AAV) capsid proteins tune and regulate the AAV infective life cycle, which can impact safety efficacy gene therapy products. Many these PTMs induce changes in protein charge heterogeneity, including deamidation, oxidation, glycation, glycosylation. To characterize heterogeneity a protein, imaged capillary isoelectric focusing (icIEF) has become gold standard method. We have previously reported an icIEF method with native...
Carbohydrate-deficient transferrin (CDT) is a reliable biomarker for chronic alcohol abuse. We developed method CDT analysis by capillary isoelectric focusing, followed immunodetection directly in the capillary, an automated fashion and on single platform (Peggy Sue™; ProteinSimple, CA, USA). Transferrin glycoforms serum samples, including disialo-transferrin, were separated their apparent points relative percentages determined. The values (percent of total transferrin) matched expected both...
<h3>Background</h3> Chimeric Antigen Receptor (CAR) T-cell therapy has revolutionized the treatment of hematologic malignancies, showing significant success against B-cell leukemia, lymphoma, and multiple myeloma. However, expanding its application to other tumors requires a deep understanding CAR signaling. Traditional methods like Western blotting flow cytometry face challenges in large-scale analyses due low throughput, high sample volumes, poor reproducibility. We achieved rapid,...
Abstract The tumor microenvironment (TME) is a complex mixture of cancerous and non-cancerous cells, including immune cells like T-cells, macrophages, neutrophils. TME plays key role in tumorigenesis metastasis, it has recently been recognized that can dramatically shape response to therapy. Thus, there pressing need accurately identify quantify the variety cell types any given TME. However, studying presents major challenges. For example, heterogeneity environment requires sensitive...