Resume

• 2005

  Ph.D.

  Biochemistry

  Organized and managed the Biochemistry/Biology Journal club\nAlberta Synchrotron Institute

  The University of Lethbridge

  Molecular simulation summer school

  Microbial Genomics and Meta-genomics

• 2000

  B.Sc (honours)

  Biochemistry

  The University of Lethbridge

• Biochemistry

  University Teaching

  Linux

  Protein Purification

  Molecular Biology

  Docking

  Microbial Ecology

  Drug Design

  Molecular Modeling

  X-ray crystallography

  Enzyme Kinetics

  Spectroscopy

  Teaching/mentoring

  Lecturing

  Problem Solving

  Protein Expression

  Supervisory Skills

  Teaching

  Microbiology

  Enzyme Assays

  Contributions of a unique Beta-clamp to substrate recognition illuminates the moleculare basis of exolysis in ferulic acid esterases

  Tim McAllister

  Chris Cote

  Lignocellulosic biomass is a promising renewable resource

  \nhowever deconstruction of this material is still the rate limiting\nstep. Major obstacles in the biocatalytic turnover of\nlignocellulose are ester-linked decorations that prevent access\nto primary structural polysaccharides. Enzymes targeting these\nesters represent promising bio-tools for increasing bioconversion\nefficiency. Ruminant livestock are unique in their ability to\ndegrade lignocellulose through the action of their gut microbiome.\nThe anaerobic fungi (phylum Neocallimastigomycota) are key\nmembers of this ecosystem that express a large repertoire of\ncarbohydrate active enzymes (CAZymes) with little sequence\nidentity with characterized CAZymes [Lombard et al. (2014)\nNucleic Acids Res. 42

  D490–D495]. We have identified a\ncarbohydrate esterase family 1 (CE1) ferulic acid esterase (FAE)\nbelonging to Anaeromyces mucronatus (AmCE1/Fae1a)

  and\ndetermined its X-ray structure in both the presence [1.55 Å\n(1 Å=0.1 nm)] and absence (1.60 Å) of ferulic acid. AmCE1\nadopts an α/β-hydrolase fold that is structurally conserved with\nbacterial FAEs

  and possesses a unique loop

  termed the β-\nclamp

  that encloses the ligand. Isothermal titration calorimetry\nreveals substrate binding is driven by enthalpic contributions

  \nwhich overcomes a large entropic penalty. A comparative\nanalysis of AmCE1 with related enzymes has uncovered the\napparent structural basis for differential FAE activities targeting\ncross-linking ferulic acid conjugates compared with terminal\ndecorations. Based on comparisons to structurally characterized\nFAEs

  we propose that the β-clamp may define the structural basis\nof exolytic activities in FAEs. This provides a structure-based\ntool for predicting exolysis and endolysis in CE1. These insights\nhold promise for rationally identifying enzymes tailored for\nbioconversion of biomass with variations in cellwall composition

  Contributions of a unique Beta-clamp to substrate recognition illuminates the moleculare basis of exolysis in ferulic acid esterases

  Tim McAllister

  Demand for meat and milk is predicted to double by 2050

  and\nmeeting this increased demand represents a “grand challenge for\nhumanity.”\n• Sustainable production practices for ruminants will require more\nefficient utilization of feed

  with a greater emphasis on the use of\nfibrous feedstuffs.\n• Fibrolytic enzyme cocktails have the potential to improve\nthe nutritional value of low quality forages

  such as straw

  and\nimprove overall feed efficiency in ruminants.\n• Available commercial fibrolytic enzymes are not specifically\ndeveloped for use in ruminant livestock and have not consistently\nimproved ruminal fiber digestion.\n• “-Omics” including

  metagenomics and metatranscriptomics

  \nhave improved our understanding of rumen microbes and the enzymes\ninvolved in deconstruction of plant cell walls.\n• A better understanding of the enzymes that limit plant cell wall\ndeconstruction in the rumen could lead to more effective fibrolytic\nenzymes additives for ruminants.

  Mining the rumen for fibrolytic feed enzymes

  Lovering AL

  Mosimann SC

  Sockett RE

  Selinger LB

  Till R

  Capeness MJ

  Thibault J

  Structural and biochemical analysis of a unique phosphatase from Bdellovibrio bacteriovorus reveals its structural and functional relationship with the protein tyrosine phosphatase class of phytase

  Tim MacAllister

  Ron Teather

  Bob Forster

  Xia Gong

  Biochemical analysis of a highly specific

  pH stable xylanase gene identified from a bovine rumen-derived metagenomic library.

  Lyn Paterson

  Ron Teather

  Tim MacAllister

  Bob Forster

  Meng Qi

  Xia Gong

  BMC Research Notes

  Cloning and identification of novel hydrolase genes from a dairy cow rumen metagenomic library and characterization of a cellulase gene.

  Tim McAllister

  Adrian Tsang

  Justin Powlowski

  Donald Patton

  Yuxi Wang

  Accepted - In Press

  Formulation of enzyme blends to maximize the hydrolysis of alkaline peroxide pretreated alfalfa hay and barley straw by rumen enzymes and commercial cellulases

  McAllister TA

  Tsang A

  Powlowski J

  Patton D

  Wang Y

  aja

  BioMed Research International

  Accepted

  Improvement in saccharification yield from rumen mixed enzymes by identification of recalcitrant cell wall constituents using enzyme fingerprinting

  Substrate binding in protein-tyrosine phosphatase-like inositol polyphosphatases

  Steve Mosimann

  Brent Selinger

  Hj Wieden

  Lisza Bruder

  Adam Smith

  Selina Dobing

  Substrate binding in protein-tyrosine phosphatase-like inositol polyphosphatases

  Elshahed MS

  McAllister T

  Forster R

  Griffith GW

  Fliegerova K

  Dagar SS

  Youssef N

  Edwards JE

  Callaghan TM

  Puniya AK

  Anaerobic fungi (phylum Neocallimastigomycota): advances in understanding their taxonomy

  life cycle

  ecology

  role and biotechnological potential

  Forster RJ

  McAllister TA

  Sensen CW

  Diversity of rumen bacteria in Canadain cervids

  L. Brent Selinger

  Steven Mosimann

  Tim Janzen

  Ralf Greiner

  Aaron Puhl

  Kinetic and structural analysis of a bacterial protein tyrosine phosphatase-like myo-inositol polyphosphatase.

  McAllister TA

  Forster RJ

  Gong X

  Biochemical and kinetic characterization of the multifunctional beta-glucosidase/beta-xylosidase/alpha-arabinosidase

  Bgxa1

  Natalie Strynadka

  Liam Worral

  Dustin King

  Journal of American Chemical Society

  New Delhi metallo-β-lactamase: structural insights into β-lactam recognition and inhibition.

  Overall CM

  Strynadka NC

  Wasney GA

  Watanabe N

  Structure of the mycosin-1 protease from the mycobacterial ESX-1 protein type VII secretion system

  Lyndsey Eltis

  Natalie Strynadka

  I Casabon

  Jenna Capyk

  Journal of Biological Chemistry

  Activity of 3-ketosteroid 9α-hydroxylase (KshAB) indicates cholesterol side chain and ring degradation occur simultaneously in Mycobacterium tuberculosis.

  Robert Forster

  Tim McAllister

  The North American Beaver (Castor canadensis) is the second largest living rodent and an iconic symbol of Canada. The beaver is a semi-aquatic browser whose diet consists of lignocellulose from a variety of plants. The beaver is a hindgut fermenter and has an enlarged ceacum that houses a complex microbiome. There have been few studies examining the microbial diversity in gastrointestinal tract of hindgut fermenting herbivores. To examine the bacterial and archaeal communities inhabiting the gastrointestinal tract of the beaver

  the microbiome of the ceacum and feaces was examined using culture-independent methods. DNA from the microbial community of the ceacum and feaces of 4 adult beavers was extracted

  and the16S rRNA gene was sequenced using either bacterial or archaeal specific primers. A total of 1447 and 1435 unique bacterial OTUs were sequenced from the ceacum and feaces

  respectively. On average

  the majority of OTUs within the ceacum were classified as Bacteroidetes (49.2%) and Firmicutes (47.6%). The feaces was also dominated by OTUs from Bacteroidetes (36.8%) and Firmicutes (58.9%). The composition of bacterial community was not significantly different among animals. The composition of the ceacal and feacal microbiome differed

  but this difference is due to changes in the abundance of closely related OTUs

  not because of major differences in the taxonomic composition of the communities. Within these communities

  known degraders of lignocellulose were identified. In contrast

  to the bacterial microbiome

  the archaeal community was dominated by a single species of methanogen

  Methanosphaera stadtmanae. The data presented here provide the first insight into the microbial community within the hindgut of the beaver.

  Bacterial and Archaeal Diversity in the Gastrointestinal Tract of the North American Beaver (Castor canadensis)

  Structure and mechanism of Staphylococcus aureus TarM

  the wall teichoic acid alpha-glycosyltransferase

  Strynadka NC

  Withers SG

  Brown ED

  Matt SolomonsonBr

  Baumann L

  Blaukopf M

  Wasney GA

  Worrall LJ

  sob

  The Proceedings of the National Academy of Sciences

  USA

  Unique to Gram-positive bacteria

  wall teichoic acids are anionic glycopolymers cross-stitched to a thick layer of peptidoglycan. The polyol phosphate subunits of these glycopolymers are decorated with GlcNAc sugars that are involved in phage binding

  genetic exchange

  host antibody response

  resistance

  and virulence. The search for the enzymes responsible for GlcNAcylation in Staphylococcus aureus has recently identified TarM and TarS with respective α- and β-(1-4) glycosyltransferase activities. The stereochemistry of the GlcNAc attachment is important in balancing biological processes

  such that the interplay of TarM and TarS is likely important for bacterial pathogenicity and survival. Here we present the crystal structure of TarM in an unusual ternary-like complex consisting of a polymeric acceptor substrate analog

  UDP from a hydrolyzed donor

  and an α-glyceryl-GlcNAc product formed in situ. These structures support an internal nucleophilic substitution-like mechanism

  lend new mechanistic insight into the glycosylation of glycopolymers

  and reveal a trimerization domain with a likely role in acceptor substrate scaffolding.

  Structure and mechanism of Staphylococcus aureus TarM

  the wall teichoic acid alpha-glycosyltransferase

  Xiying H

  McAllister T

  Vederas D

  Yanke J

  Gilroyed B

  Nkemka V

  Bioresources Technology

  Accepted

  Bioaugmentation with an anaerobic fungus in a two-stage process for biohydrogen and biogas production using corn silage and cattail

  Steven C. Mosimann

  L. Brent Selinger

  Effect of ionic strength and oxidation on the P-loop conformation of the protein tyrosine phosphatase like phytase

  PhyAsr.

  Steven C. Mosimann

  L. Brent Selinger

  Structural Analysis of a Multifunctional

  Tandemly Repeated Inositol Polyphosphatase

  Gruninger

  University of Lethbridge

  Agriculture and Agri-Food Canada

  The University of British Columbia

  University of Lethbridge

  Lethbridge

  Alberta

  Taught an introductory microbiology course during the fall semester of 2013 at the University of Lethbridge that focused on both the practical and clinical aspect of medical microbiology. Prepared

  administered

  and marked assignments and exams. Assigned final grades.

  HLSC2400 - Medical Microbiology - Sessional Instructor

  University of Lethbridge

  Lethbridge

  AB

  Research Scientist using -omic techniques

  microbiology

  protein biochemistry and animal sciences to examine the role of the gut micrbiome on the health and efficiency of beef cattle.

  Research Scientist

  Agriculture and Agri-Food Canada

  Structural analysis of bacterial cell wall enzymes

  structure based drug design

  biophysical analysis of protein ligand interactions (ITC

  CD

  UV/VIS/fluorescence spectroscopy)

  molecular biology

  protein expression and purification

  The University of British Columbia

  Agriculture and Agri-Food Canada

  Lethbridge Research Center

  Rumen microbiology

  microbial ecology

  transcriptomics

  enzyme structure/function

  Research Associate

  Delivered lectures on introductory biochemistry. Course covered introduction to Amino acids and proteins

  lipids

  carbohydrates and metabolism. Prepared

  administered and marked assignments and exams. Assigned final grades.

  University of Lethbridge

  Lecturer (Enzymology)

  Delivered lectures on enzyme structure and mechanism

  methods to probe enzyme mechanism

  and structure based drug design to a 4th year enzymology class (130 students/year). Prepared

  administered

  and marked assignments and exams

  The University of British Columbia

  Michael Smith Foundation for Health Research Postdoctoral Fellowship

  Michael Smith Foundation for Health Research

  NSERC Government Labs Visiting Fellowship

  Natural Sciences and Engineering Research Council of Canada