The research in this laboratory focuses on the enzymology, pathways and physiology of microorganisms that degrade environmental contaminants. The organisms we study are primarily hydrocarbon-oxidizing strains that degrade compounds such as trichloroethylene (TCE), methyl tertiary butyl ether (MTBE) and 1,4-dioxane (14D). The approaches used in this laboratory range from the isolation and characterization of novel strains through to genomic and proteomic analyses of individual strains and microbial communities. Many of these approaches make use of stable isotopes. Our goal is to understand the mechanisms utilized by naturally occurring microorganisms to degrade contaminants and to use this information to help develop appropriate treatment strategies that maximize the activities of these microorganisms in contaminated environments.
MBA Oregan State University 2001
Ph.D. Biochemistry University of Bristol, UK 1985
B.S. Botany University College, University of London, UK 1980
Area(s) of Expertise
Bioremediation, Microbial Physiology, Enzymology, Environmental Microbiology
- Draft Genome Sequences of Four Aerobic Isobutane-Metabolizing Bacteria , MICROBIOLOGY RESOURCE ANNOUNCEMENTS (2021)
- Long-term cometabolic transformation of 1,1,1-trichloroethane and 1,4-dioxane by Rhodococcus rhodochrous ATCC 21198 grown on alcohols slowly produced by orthosilicates , JOURNAL OF CONTAMINANT HYDROLOGY (2021)
- Co-encapsulation of slow release compounds and Rhodococcus rhodochrous ATCC 21198 in gellan gum beads to promote the long-term aerobic cometabolic transformation of 1,1,1-trichloroethane, cis-1,2-dichloroethene and 1,4-dioxane , Environmental Science: Processes & Impacts (2020)
- 1,4-Dioxane in drinking water: Emerging for forty years and still unregulated , Current Opinion in Environmental Science & Health (2019)
- Aerobic cometabolism of 1,4-dioxane by isobutane-utilizing microorganisms including Rhodococcus rhodochrous strain 21198 in aquifer microcosms: Experimental and modeling study , SCIENCE OF THE TOTAL ENVIRONMENT (2019)
- Nitrogen Gas Fixation and Conversion to Ammonium Using Microbial Electrolysis Cells , ACS Sustainable Chemistry & Engineering (2019)
- Biodegradation of Ether Pollutants , Consequences of Microbial Interactions with Hydrocarbons, Oils, and Lipids: Biodegradation and Bioremediation (2018)
- Concurrent Treatment of 1,4-Dioxane and Chlorinated Aliphatics in a Groundwater Recirculation System Via Aerobic Cometabolism , Groundwater Monitoring & Remediation (2018)
- Enrichment with Carbon-13 and Deuterium during Monooxygenase-Mediated Biodegradation of 1,4-Dioxane , Environmental Science & Technology Letters (2018)
- Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea , Appl. Environ. Microbiol. (2016)
The project objective is to use apply a recently-developed activity-based labeling (ABL) technique to detect, and identify contaminant-degrading monooxygenase enzymes expressed by native or augmented microorganisms. The 2-step technique involves an initial inactivation of target enzymes using diyne probes with subsequent use of copper-catalyzed alkyne/Azide cycloaddition (CuAAC) reaction which generates a fluorescent protein adduct. This adduct can be detected and quantified by a number of different analytical approaches such as SDS-PAGE, microscopy, and flow cytometry. The technical objective of the proposed work will be to use ABL approaches to support a concurrent but separately funded study that aims to demonstrate that in situ aerobic cometabolic treatment of dilute plumes of chlorinated volatile organic compounds (CVOC)s can be achieved using bacteria grown on substrates including 2-butanol and benzyl alcohol. The ABL technique is expected to provide data that will enable the accompanying project to confirm the role of specific monooxygenases in in situ contaminant biodegradation and to determine the abundance of bacteria expressing these specific monooxygenases.
The overall aim of this project is to evaluate the use of aerobic alkane-oxidizing bacteria for the in situ cometabolic degradation of 1,4-dioxane (14D). The project will involve testing field samples for the stimulation of either indigenous gaseous alkane and alkyne-metabolizing bacteria, testing the potential for bioaugmentation of field samples and detecting the presence of active alkane-oxidizing bacteria in field samples using activity-based labeling of catalytically active monooxygenases.
The alkane-oxidizing bacterium Rhodococcus rhodochrous ATCC 21198 can degrade 1,4-dioxane (14D) at high rates for over 300 days when it is co-encapsulated in gellan gum with orthosilicate slow release compounds (SRCs) that hydrolyze to produce alcohols. The overall goal of this project is to further develop this co-encapsulation technology for passive and sustainable aerobic cometabolic systems for the treatment of emerging contaminants such as N-nitrosodimethylamine (NDMA), 1,2,3-trichloropropane (TCP), as well as important contaminant mixtures such as 14D and its common co-contaminants, 1,1,1 trichloroethane and cis-dichloroethene.The contaminant-degrading activity of ATCC 21198 is due to non-specific, alkane-inducible monooxygenases that normally function to initiate alkane catabolism. In the absence of alkanes, the factors that control expression of these monooxygenases and enable sustained contaminant degradation are unknown but are key to further developing the co-encapsulation technology. Activity measurements, activity-based monooxygenase labeling and whole cell proteome analyses will be used to separately characterize the effects of alcohols, SRCs, starvation, encapsulation, and non-growth supporting contaminants on expression of monooxygenases and other enzymes in strain ATCC 21198. The physiological and enzymatic changes that occur the 300-day life cycle of this strain when co-encapsulated with model SRCs will also be determined. Similar genome-enabled approaches will also be used to identify other pure cultures with alternative monooxygenase complements that can sustainably degrade chlorinated ethenes (trichloroethene, vinyl chloride and 1,1-dichloroethene), and emerging contaminants (NDMA, and TCP) when co-encapsulated with SRCs. The activities of the co-encapsulated strains will be tested in batch reactors containing beads with single cultures and SRC as well as bead mixtures with different cultures and SRCs.
Fluorescence activated cell sorting (FACS) is a technique that involves sorting away select cells (or objects) from complex mixtures based on their intrinsic or acquired fluorescence. FACS is a transformative technology that allows the study of the unculturable microbes (which are numerically dominant in nature) and accomplish tasks that are highly laborious or impossible complete in other ways; the technology has led to significant discoveries in many microbial research fields (e.g. ecology, genetics, physiology, symbiosis/interactions, bioengineering, and bio-prospecting), and nearly single-handedly forged new fields of research, e.g. single cell genomics and transcriptomics, which involves the study of DNA and mRNA from individual cells. More than 25 North Carolina State University (NCSU) faculty, belonging to 4 colleges, have needs for FACS in their research or teaching programs; however, NCSU lacks a FACS instrument optimized for the analysis of non-mammalian microbial cells (e.g. bacteria, archaea and fungi), and to our knowledge, no â€˜microbe optimizedâ€™ instrument is available at research universities within the Research Triangle of North Carolina (e.g. UNC-CH, Duke University). Here, funds are requested to acquire a Becton Dickinson FACSMelody flow cytometer, a versatile (3 excitation laser, 9-color detection) and â€˜turn keyâ€™ system, which fundamentally enables microbiological research that is highly laborious or impossible to accomplish without it. The FACSMelody is powerful yet simple to use and generates easy to grasp visual (flow cytometric) data â€“ making it a good potential training and educational tool for undergraduate/graduate courses and workshops. A FACSMelody system is ideal for getting FACS technology rapidly and easily into the hands of faculty in need. Overall, a FACS system for non-mammalian microbial research is needed for NCSU to be innovative, internationally competitive at attracting new faculty and highly talented students, and foster creative future proposals.
The key objective of this project is to demonstrate that a novel multiple primary substrate (MPS) cometabolic biosparging technology can meet DoD needs for reliable, flexible, and cost effective treatment of a groundwater with co-mingled 1,4-dioxane (14D) and chlorinated volatile organic compounds (CVOCs). Isobutane will be added to the aquifer to target 14D biodegradation, and methane, isobutylene, and/or propane will be added to stimulate degradation of trichloroethene (TCE) and other CVOCs present. The ability of the technology to achieve regulatory levels of 14D and CVOCs in the groundwater, and the cost of the technology will be evaluated.