Progress 01/01/18 to 12/31/22
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: Scientific peer-reviewed journal articles. Presentations at scientific meetings. Postdoc and graduate student training Changes/Problems:We have completed the proposed work. However, based on the findings fromchicken trials, which were not as good as expected, we speculated the bioavailability of the hydrophobic BSH inhibitors was low if we just directly supplement the feed with the BSH inhibitors; thus, in vivo bioavailability and functionality of orally administered BSH inhibitorcould be greatly enhanced using cost effective encapsulation technology. Therefore, we also devoted to developing and optimizing an effective encapsulation technology that aws not proposed in this project. We have successfully optimized and validated a cost-effectiveencapsulation technology, andprepared large quantities of encapsulated CAPE for a large chicken trial in the future. What opportunities for training and professional development has the project provided? USA. A Ph.D student was supported by this grant to perform chicken trials to assess three BSH inhibitors. This graduate student, supervised by both Dr. Wei Zhai (Co-PI) and Dr. Jun Lin (PI), has received comprehensive education training and obtained hands on experience on chicken nutrition, physiology, and gut microbiota. In addition, this project also partly supported a Postdoc to gain training in the areas of molecular microbiology, bioinformatics, and enzymology; this postdoc also coordinated with the scientists in Ireland and Northern Ireland for successful completion of this transdisciplinary project. Finally, this project supported Dr. Jun Lin (PI) to attend national and international meetings to disseminate science-based information to public. Republic of Ireland. This project provide Dr. Susan Joyce (Co-PI) opportunities to hire Postdoc and graduate student to perform basic and translational studies related to BSH enzymes. With aid of this project, Dr. Joyce and her trainees also have been involved in various professional development and outreach activities, such as attending Science Foundation IrelandScience Policy Summit (November, 2019), Profile and short research snapshop to celebrate IBD day - twitter and Facebook (December, 2019), hosting Transition Year students for Young Scientist Project in order to compete in all-Ireland young scientist competition (December, 2019), giving banquet talk focused on the Theme: food for health(June, 2019, Cornstore, Limerick). Northern Ireland. This project provide Dr. Irina Tikhonova(Co-PI) opportunities to hire a Postdoc fellowto perform the computational aspect of the project.Dr. Warispreet Singh, the first postdoc, had an opportunity to attend the CCPBioSIM training week, May 20-24, 2019 at the University of Bristol, where he has received training in current computational methodologies and tools (Python, Jupyter Notebooks, BioSimSpace, Free energy calculations and, etc.). Waris has also attended a number career and professional workshops and courses at Queen's University Belfast to improve his writing skills, to learn more about time management and leadership. Drs. Tikhonova and Singh have hosted and trained summer students supported by IAESTE and Nuffield Foundation in computer simulations of BSH enzymes (June-August, 2019). Following leaving of Dr. Singh, the new PDRA, Dr. Dmitry Karlov, was hired on October 1, 2020 during COVID 19 pandemics. Dr. Dmitry Karlov has been actively working on this project and was engaged in extensivepost-doctoral training programs provided by QUB,participated in a number of training sessions provided by Queen's University Belfast such as writing for publication, developing research carrier, developing CV, networking and profile building for researchers. How have the results been disseminated to communities of interest?Despite the challenging COVID-19 pandemic in 2020-2022, we have continued to identify various opportunities to disseminate our results to communities of interest. Probiotics, Prebiotics &New food -Symposium 13th September 2021 (Keynote address) Microbiology Society UK and Ireland 28th April 2021 (Plenary speaker) Irish Mass Spectrometry Meeting May 2021 (post doc presentation) 4th CCPBioSim/CCP5 Multiscale Modelling Conference on 29-31 March 2021 (presentation) What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
With support by this Tri-Partite NIFA project, our international collaborative team has made significant progresses toward development of innovative alternatives to antibiotic growth promoters by targeting bile salt hydrolase (BSH), a gut microbiome target that plays a critical role in host energy harvest. Briefly, this project has led to 10peer-reviewed high-impact journal articles (8 published and 2 under review), 7 abstracts presented in national and international conferences, and 2 PhD dissertations. Objective 1. Evaluate identified BSH inhibitors in a well-simulated chicken experimental system. Threebroiler chicken trials were conducted (USA) and analysed for physiological effects on BA signatures (ROI) as well as other microbial and host physiology outcomes (USA). Three papers were published from two trials. For the last trial, two attempts to deliver the material (chicken feces and blood) from USAto ROI was impeded by transportation and customs issues (due to Brexit and to COVID). The samples eventually received by ROI had lost their integrity due to conditions of storage. Based on the findings fromchicken trials, which were not as good as expected, we speculated the bioavailability of the hydrophobic BSH inhibitors was low if we just directly supplement the feed with the BSH inhibitors; thus, in vivo bioavailability and functionality of orally administered BSH inhibitorcould be greatly enhanced using cost effective encapsulation technology. Subsequently, we also devoted to developing and optimizing an effective encapsulation technology. We have successfully optimized and validated a cost-effectiveencapsulation technology, andprepared large quantities of encapsulated CAPE for a large chicken trial in the future. Objective 2. Understand the mechanistic basis of BSH inhibitors through analysis of ex vivo samples. Substrate specificity has been delineated using a multidisciplinary approach including mass-spectrometry, mutagenesis, molecular dynamic simulations, machine learning, and crystallography. L. salivarious BSHwas chosen and characterized for exact modifications to BAs by Mass spectrometry (MS, ROI). Based on these interactions and substrate preferences, our NI partner predicted a series of residues (25 total) for mutation to alter or reinstate functionality (double mutations). These plasmidic constructs were sent to ROI partner to interrogate gain or loss of activity. ROI interrogated the microbiome representations, to decipher BSH classes based on activity and genomic representation of BSHs. Cloning, expression of recombinant BSH (rBSH) and characterization of their activity was conducted using co-incubation assays and MS applications. The existing inhibitors (CAPE, Carnosic acid, Riboflavin) are subsequently shown to alter each class selectively and in a substrate-enzyme dependent manner. Michaelis menton kinetics with each class indicate substrate specific affinity and velocity. This work is being extended, following further funding acquisition (post doctoral fellowship) to understand these kinetic interactions and inhibitor effects on them, in order to bring this work to publication level for all 3 partners. Objective 3. Understand BSH structure-function relationships and structure-based design of BSH inhibitors. A series of 23 BSH enzymes were selected for investigations based on their sequence and classication from previous publications. The diverse BSH enaymeshave been examined for functional activity (Joyce, RoI),structure-function (Tikhonova, NI), critical role of specific residues in BSH function using amino acid-substitution mutagenesis(Lin, USA). Extensive structure-function studies have been performed on the Lactobacillus salivarius BSH under active and seamless collaborations among Lin (PI, USA), Tikhonova (Co-PI, NI), and Joyce (Co-PI, RoI), which has led to a panel of high-impact journal articles and abstracts presented in conferences. Virtual screening of several natural product databases (Drugbank (4,000 compounds), ZINC natural products (225,000 compounds), Coconut natural products (400,000 compounds)) has been completed by a new PDRA. The virtual screening workflow has included docking of compounds in a set of LsBSH conformations obtained from molecular dynamics (MD) simulation studies, selection of the best 100 compounds and evaluation of these compounds using MD simulations and molecular mechanics, Poisson-Boltzmann Surface Area (MM-PBSA) calculations (Tikhonova, Northern Ireland). 32 purchasable compounds have been selected for experimental validation (Joyce, Ireland) and six inhibitors were identified. The identified new BSH inhibitors are attractive non-antibiotic growth promoters for enhanced animal production and sustainability.
Publications
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2018
Citation:
Wenjing Geng, 2018. Bile salt hydrolase: from basic science to translational innovation. Ph.D. Dissertation
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2018
Citation:
Geng, W., C. Gahan, S. Joyce, J. Chang, A. Saxton, J. Lin. 2018. In vivo evaluation of bile salt hydrolase inhibitors using
broiler chicken. American Society for Microbiology Annual Meeting. June 7-11, Atlanta, GA
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Lin, J., W. Geng, S. Long, S. Joyce. 2018. Improved animal husbandry through inhibition of gut microbial bile salt
hydrolase. Focused Meeting 2018: Microbiomes Underpinning Agriculture. October 1-2, Cork, Ireland
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Xu, F., X.J. Hu, W. Singh, W. Geng, I. Tikhonova, J. Lin. 2019. The complex structure of the bile salt hydrolase from Lactobacillus salivarius reveals the residues contributing to catalysis and substrate specificity. Scientific Reports. 9:12438.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Prete, R., Long S.L., , Lopez Gallardo, A., Gahan, C., Corsetti, A. & S.A. Joyce. 2020. Beneficial bile acid metabolism from
Lactobacillus plantarum of food origin. Scientific Reports 10, 1165 (2020). https://doi.org/10.1038/s41598-020-58069-5
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Geng, W., S. Long, Y.J. Chang, A. Saxton, S. Joyce, J. Lin. 2020. Evaluation of in vivo efficacy of bile salt hydrolase inhibitors using chicken model system. Scientific Reports. 10(1):4941. DOI: 10.1038/s41598-020-61723-7
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J. 2019. Sustainability and safety of animal agriculture: emerging threats and innovative solutions. The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products. Ningbo, China, November 16-18, 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Zhang, B., and W. Zhai. 2019. Effects of riboflavin on growth performance, processing yield, and internal organ development of Ross 708 male broilers. Poult. Sci. 98 (E-Suppl. 1). Page 27. Abstract # M84. 2019 International Poultry Scientific Forum in Atlanta, GA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Singh W, Joyce SA, Lin J, Tikhonova IG. 2019. Structure-based Search of Bile Salt Hydrolase Inhibitors. 7th Annual CCPBioSim Conference Frontiers in Bimolecular Simulations (September 4 -6, 2019, Bristol).
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Sarah L. Long. 2019. Ph.D. dissertation. APC Microbiome Ireland, School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland. Ph.D. Mentor: Dr. Susan A. Joyce.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Poudel, S., L. Zhang, G.T. Tabler, J. Lin, W. Zhang. 2021. Effects of riboflavin and Bacillus subtilis on internal organ development and intestinal health of Ross 708 male broilers with or without coccidial challenge. Poultry Science. 100:100973.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Poudel, S., G.T. Tabler, J. Lin, W. Zhai, L. Zhang. 2022. Riboflavin and Bacillus subtilis effects on growth performance and woody-breast of Ross 708 broilers with or without Eimeria spp. challenge. Journal of Animal Science and Technology. 64(3): 443461
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Joyce and O'Malley. 2022. Bile acids, bioactive signalling molecules in interoceptive gut-to-brain communication. Journal of Physiology. 600:2565-2578
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Dmitry, S. K., S. L Long, X. Zeng, F. Xu, K. Lal, L. Cao, K. Hayoun, J. Lin, S. A. Joyce, I., Tikhonova. 2023. The mechanism of substrate specificity in bile salt hydrolase from experiment and computation. Structure Cell Press (In Press)
- Type:
Journal Articles
Status:
Submitted
Year Published:
2023
Citation:
Long and Joyce. 2023. Rational selection and Characterisation of bile acid (BA) metabolising species of infant origin. OPEN ACCESS DEPOSITION: bioRxiv 2022.06.24.497474; doi: https://doi.org/10.1101/2022.06.24.497474 Submitted to Nutrients
- Type:
Journal Articles
Status:
Submitted
Year Published:
2023
Citation:
Dmitry S. Karlov, Sarah L. Long, Kanhaya Lal, Karim Hayoun, Jun Lin, Susan A. Joyce and Irina G. Tikhonova.2023. Virtual screening and validation of novel Bile Salt Hydrolase inhibitors. Scientific Reports
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Dmitry S. Karlov,, Susan A. Joyce, Jun Lin, and Irina G. Tikhonova. 2021. Guiding the Bile Salt Hydrolase activity landscape exploration with molecular modelling and supervised machine learning. The 7th Annual CCPBioSim Conference. University of Edinburg.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Dmitry S. Karlov,, Susan A. Joyce, Jun Lin, and Irina G. Tikhonova. 2022. Delineating Bile Salt Hydrolase selectivity using machine learning and MD simulations. The 8th Annual CCPBioSim Conference. University of Edinburg.
|
Progress 01/01/21 to 12/31/21
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A postdoctoral researcher in PI's lab has been focused on purification of large quantities of BSH for structural and functional work. This postdoc was also in charge of all microbiome analysis. A PDRA working on this project, was engaged in the post-doctoral training programs provided by QUB and focused on the opportunities for the industrial application of the academical research results leading by Elasmogen Ltd. A postdoctoral scientist was engaged to the programme. COVID interrupted work and rota system for 50% Laboratory occupancy slowed and impacted progression as well as contraction of COVID by different Laboratory members. How have the results been disseminated to communities of interest?Despite the challenging COVID-19 pandemic in 2021, we have continued to identify various opportunities to disseminate our results to communities of interest. Probiotics, Prebiotics &New food -Symposium 13th September 2021 (Keynote address) Microbiology Society UK and Ireland 28th April 2021 (Plenary speaker) Irish Mass Spectrometry Meeting May 2021 -1 (post doc presentation) 4th CCPBioSim/CCP5 Multiscale Modelling Conference on 29-31 March 2021 (presentation) What do you plan to do during the next reporting period to accomplish the goals? To analyze the results of virtual screening and to optimize found hits. To analyze the results of the last mutagenesis data using MD simulations. To prepare three drafts of the manuscript for publication.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate identified BSH inhibitors in a well-simulated chicken experimental system. We have evaluated the effect of BSH inhibition (n=4 inhibitors) through feed (published) and through dose dependent determination assays (manuscript in preparation) in heterogenetic chickens. Our data reveals that the effective dosage for chicken responder and non-responder populations to specific inhibitors, is in the range for riboflavin (6.6-20 ppm), for CAPE (10-20 ppm) and for carnosic acid (10-20 ppm) on the basis of alterations to bile acid signatures and moieties indicative of concrete but differential degrees towards weight gain. The Lin laboratory (USA) have conducted corresponding microbial assessment. Our combined work identified the need for precision delivery. US researchers developed efficient encapsulation methodology. Subsequently, a follow up trial and microbial assessment was undertaken. Samples shipped (twice, were massively delayed, in delivery, due to Brexit and COVID. They were therefore ineligible for interrogation. Our NI partner identified 30+ potential inhibitors of LsBSH. Based on energetics of reactions 10 of these were selected for activity determination against purified LsBSH protein. This work is on-going. (Joyce, Ireland & Lin, USA) Objective 2. Understand the mechanistic basis of BSH inhibitors through analysis of ex vivo samples. Crystal structure indicated that deconjugation of Taurine conjugated bile acids may result in a metabolite of taurine, 2-amino-2-hydroxyethane-1-sulfonic acid (TAW), fixed to L. acidophilus BSH enzyme. We (ROI) established specific detection methods for TAW and associated taurine metabolites and performed assays with free taurine as well as Taurine bound to bile acids as substrates. Our initial indications are that specific classes (7 clades represented, 23 BSHs were cloned and tested for actual activity) of BSH show a slight elevation of TAW when plasmid bourne BSH is applied to assay. Residual activity was also a feature of the plasmid containing strains. BSH protein purification, representing our in silico identified 7 BSH gut activity clades or classes were protein purified. Further work is underway with these purified BSHs to determine if TAW is a consequence of BSH activity. (Joyce, Ireland & Lin, USA) The cloned and tested 23 BSHs representing just 7 clades of gut resident BSHs, have different degrees of specificity for bile acid substrates. These proteins have been purified and are subject to Michaelis Menton kinetic analysis towards identifying best substrates for each. Our hypothesis here is that different gut representatives will be differentially susceptible to BSH inhibitors and that they may identify bespoke inhibition targeted towards weight gain in a class and clade dependent manner. Laboratory work is ongoing, with inhibitors and purified rBSHs to determine specificity. (Joyce, Ireland & Lin, USA) Objective 3. Understand BSH structure-function relationships and structure-based design of BSH inhibitors. The analysis of molecular dynamic (MD) simulations in conjunction with mutagenesis data has been performed. To accomplish this task a regression analysis was built based correlating activity variation and structural changes of the enzyme binding site. The relative importance of the regression coefficients regression allowed us to identify the region of the binding site with the highest influence on the activity. We performed long MD simulations of several covalent complexes - mutant/substrate representing the rate-limiting step of the enzyme hydrolysis. Hydrogen bonding and water thermodynamics analysis allowed us to explain the major trends in substrate specificity. The manuscript is on the final stage (Tikhonova, Northern Ireland) MD simulations performed on ARCHER2 were used for the mapping of the LsBSH active site using the Site Identification by Ligand Competitive Saturation (SILCS) technique. Since the information about the BSH inhibitors is limited, we performed cosolvent MD simulations to free energy maps for probes of different nature: aromatic, aliphatic, hydrophilic, charged. Based on these maps a protein specific scoring function has been derived and will be used for optimization of the inhibitors identified in virtual screening. (Tikhonova, Northern Ireland). We identified key potential residues of interaction for LsBSH with individual conjugated bile acid moieties. From here, 25 different mutations were predicted to alter activity of LsBSH. Lin (USA) performed mutation of plasmid borne LsBSH and they were analysed for activity, interaction with and accumulation of bile acid moieties (figure 1 below). The indications are that predictions confirmed either reduced, killed, or enhanced activity of LsBSH. (Tikhonova, Northern Ireland, Joyce, Ireland & Lin, USA)
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2022
Citation:
Sabin, P., G.T. Tabler, J. Lin, W. Zhai, L. Zhang. Riboflavin and Bacillus subtilis effects on growth performance and woody-breast of Ross 708 broilers with or without Eimeria spp. challenge. Journal of Animal Science and Technology
- Type:
Journal Articles
Status:
Submitted
Year Published:
2022
Citation:
Joyce, SA et al. Bile acids, bioactive signalling molecules in interoceptive gut-to-brain communication. Journal of Physiology
|
Progress 01/01/20 to 12/31/20
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles. • Presentations at scientific meetings. • Postdoc and graduate student training Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? USA. A graduate student at Mississippi State University was supported by this grant to perform large dosimetric chicken trials to assess three BSH inhibitors. This graduate student, supervised by Dr. Wei Zhai (Co-PI) and Dr. Jun Lin (PI), has received comprehensive education training and obtained hands on experience on chicken nutrition, physiology, and gut microbiota. This student obtained M.S. degree in spring 2020. In addition, this project also partly supported a research scientist Dr. Ximin Zeng in PI's lab to gain training in the areas of molecular microbiology, bioinformatics, and enzymology; tDr. Zeng also coordinated with the scientists in Ireland and Northern Ireland by performing amino acid mutagenesis research. Northern Ireland. The first PDRA, Warispreet Singh has resigned on 12/2019. Due to COVID19, there was a delay in arrival of the new PDRA, Dmitry Karlov. He has arrived on October 1, 2020 instead of March 2020. Nevertheless, in the past three months he has completed several QUB online trainings and career workshops. Republic of Ireland. Dr Lekha Menon Margassery resigned to return to family during COVID (September 2020) Dr Karim Hayoun has been appointed and he began his work in January this year. A No cost extension has been requested, to DAFM (Joyce, Ireland). Training was not performed outside health and safety during this time How have the results been disseminated to communities of interest?Due to COVID-19 pandemic in 2020, we only had limited activities to disseminate our results to communities of interest. For example, the results of computational work were used to prepare a HECBioSIM-ARCHER2 application to request 10,776 kAU computational time on the National Supercomputing facility - ARCHER2 in order to further run BSH MD simulations. The application has been successful. What do you plan to do during the next reporting period to accomplish the goals? We plan to perform a large chicken trial with comprehensive nutrition, physiology, microbiome, and metabolome studies as proposed in Objective 1 and 2. For this trial, we will include a new treatment in which feed will be supplemented with encapsulated BSH inhibitor (Lin/Zhai, USA & Joyce, Ireland). We will carefully analyze generated MD simulation trajectories of BSH in various forms to further understand the catalytic mechanism and substrate specificity. We plan to generate two manuscripts for peer-reviewed journals. One is around substrate specificity in LsBSH and another one is around isotype specificity (Tikhonova, Northern Ireland; Lin, USA; Joyce, Ireland) We will analyze the results of experimental test for virtual screening and suggested mutations to further explore possibility to inhibit BSH isotypes (Tikhonova, Northern Ireland; Lin, USA; Joyce, Ireland) We will investigate inhibitor action through an alternative delivery route to animals and study their specific interaction in terms of metabolites, lipids and microbiome influences (Lin, USA; Joyce, Ireland) We will determine, to publication standard, the impact of new BSH inhibitors on LsBSH (Joyce, Ireland; Tikhonova, Northern Ireland; Lin, USA). We will investigate the downstream generation of Taw as a function of BSH activity (Joyce, Ireland; Lin, USA).
Impacts
What was accomplished under these goals?
Objective 1. Evaluate identified BSH inhibitors in a well-simulated chicken experimental system. An initial longitudinal analysis for dose inhibition was performed (Lin, USA) with samples shipped and analysed for bile acid metabolism (Joyce, UCC, Ireland) and their potential and actual receptor activation (Joyce Lab, Ireland; Lin, USA). Responders and non-responders in the population were identified, this work is published https://doi.org/10.1038/s41598-020-61723-7. A dose dependency study was performed and analysed for efficiency over time to reveal optimal dosage for bile acid modification and weight alteration. This work is in preparation for publication (Lin, USA). We completed the proposed study to evaluate the interaction of BSH inhibitor and probiotics. Specifically, we investigated the effects of riboflavin and Bacillus subtilis on growth performance, internal organ development and intestinal health of broilers with or without coccidial challenge. This study has led to one paper published on Poultry Science (Zhai/Lin, USA) After we optimized and validated an encapsulation technology, we prepared large quantities of encapsulated CAPE. The encapsulated CAPE together with CAPE as well as antibiotic growth promoters will be evaluated in a large and comprehensive chicken trial in 2021 (Lin, USA). Objective 2. Understand the mechanistic basis of BSH inhibitors through analysis of ex vivo samples. BSH sequence analysis and extensive molecular dynamics (MD) simulations of lsBSH crystal structures in substrate-bound and unbound forms were conducted to further study the catalytic mechanism of bile salt hydrolysis in lsBSH. His140 and Glu153 have been selected for mutagenesis studies to probe their role in assisting the catalytic process by stabilizing the zwitterionic form of Cys2 (Tikhonova, Northern Ireland). The experimental work involving mutant construction and UPLC-MS substrate profiling is completed and indicates loss of function, selective loss of function and modulation for interaction with specific bile acids (Joyce, Ireland & Lin, USA) A quantitative model of substrate binding that describes the relationship between variation in substrate binding and residue interaction energy has been built based on UPLC-MS data and free energy calculations (Tikhonova, Northern Ireland, Joyce, Ireland & Lin, USA). The model is now used to interpret the results of mutations and the preference of TCA binding by LsBSH. The rational is built on the basis of the new LsBSH crystal structure bound to TCA (Lin, USA). The manuscript is in preparation. A metabolic intermediate was identified (TAW) in initial cystal structures of lsBSH (Lin, USA). Different approaches and method development were applied to detect TAW and to examine whether BSH was connected to its preparation. Taw intermediates were detected in the presence of specific tauro-conjugated bile acids (Joyce, Ireland). Further investigations with specific purified proteins representing different clades, are underway in order to determine whether this functional metabolite can be assigned o BSH activity (Joyce, Ireland). A series of 23 BSH were selected for investigations based on their sequence and classification from the following publications: Jones et al., 2008 PNAS, doi: 10.1073/pnas.0804437105; Liang et al. (2018) doi: 10.3390/molecules23051157; Song et al., (2019), doi: 10.1186/s40168-019-0628-3. They are characterized in silico and for their exact functional activity determined and stratified (Joyce, Ireland). They are currently being interrogated in structure-function assignment (Tikhonova, Northern Ireland). Their representative proteins have been purified and are currently being tested as 5 representative clades and functionalities with specific BSH inhibitors (Joyce, Ireland). Objective 3. Understand BSH structure-function relationships and structure-based design of BSH inhibitors. Virtual screening of several natural product databases (Drugbank (4,000 compounds), ZINC natural products (225,000 compounds), Coconut natural products (400,000 compounds)) has been completed by a new PDRA. The virtual screening workflow has included docking of compounds in a set of LsBSH conformations obtained from molecular dynamics (MD) simulation studies, selection of the best 100 compounds and evaluation of these compounds using MD simulations and molecular mechanics, Poisson-Boltzmann Surface Area (MM-PBSA) calculations (Tikhonova, Northern Ireland). 32 purchasable compounds have been selected for experimental validation (Joyce, UCC, Ireland). Understanding of similarities and differences between BSHs from different isotypes. Multiscale MD simulations of five BSH isotypes with available crystal structures in the GCA and TCA-bound and unbound forms have been completed (Tikhonova, Northern Ireland). Currently, we perform careful analysis of the generated MD trajectories in conjunction with the results of mutagenesis (Joyce, Ireland & Lin, USA) to establish structural difference in substrate specificity. The manuscript is in preparation.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2020
Citation:
Poudel, S., L. Zhang, G.T. Tabler, J. Lin, W. Zhang. 2020. Effects of riboflavin and Bacillus subtilis on growth performance and woody breast condition of Ross 708 male broilers with or without coccidial challenge. Poultry Science
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Poudel, S., L. Zhang, G.T. Tabler, J. Lin, W. Zhang. 2021. Effects of riboflavin and Bacillus subtilis on internal organ development and intestinal health of Ross 708 male broilers with or without coccidial challenge. Poultry Science. 100:100973.
|
Progress 01/01/19 to 12/31/19
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • Scientific peer-reviewed journal articles. • Presentations at scientific meetings. • Postdoc and graduate student training Changes/Problems:Based on the findings from three large chicken trials, which were not as good as expected, we speculated the bioavailability of the hydrophobic BSH inhibitors could be enhanced using cost effective encapsulation technology. Thus, we brouhgt a new approach in this project that was not proposed in original application. Specifically, we devoted to developing and optimizing an effective encapsulation technology that can greatly improve in vivo availability and functionality of BSH inhibitor. Due to budget limit, in the future, we will primarily focus on in vivo assessment of the most promising and feasible BSH inhibitor CAPE. What opportunities for training and professional development has the project provided? USA. A Ph.D student at Mississippi State University was supported by this grant to perform large dosimetric chicken trials to assess three BSH inhibitors. This graduate student, supervised by Dr. Wei Zhai (Co-PI) and Dr. Jun Lin (PI), has received comprehensive education training and obtained hands on experience on chicken nutrition, physiology, and gut microbiota. This Ph.D. student is expected to obtain Ph.D. in spring 2020. In addition, this project also partly supported a Postdoc to gain training in the areas of molecular microbiology, bioinformatics, and enzymology; this postdoc also coordinated with the scientists in Ireland and Northern Ireland for successful completion of this transdisciplinary project. Finally, this project supported Dr. Jun Lin (PI) to attend the First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products held in Ningbo, China (November 16-18, 2019). Republic of Ireland. This project provide Dr. Susan Joyce (Co-PI) opportunities to hire Postdoc and graduate student to perform basic and translational studies related to BSH enzymes. With aid of this project, Dr. Joyce and her trainees also have been involved in various professional development and outreach activities, such as attending Science Foundation Ireland Science Policy Summit (November, 2019), Profile and short research snapshop to celebrate IBD day - twitter and Facebook (December, 2019), hosting Transition Year students for Young Scientist Project in order to compete in all-Ireland young scientist competition (December, 2019), giving banquet talk focused on the Theme: food for health (~100 diners) (June, 2019, Cornstore, Limerick) Northern Ireland. Dr. Warispreet Singh, a research fellow hired to perform the computational aspect of the project had an opportunity to attend the CCPBioSIM training week, May 20-24, 2019 (http://www.ccpbiosim.ac.uk/events/workshop-course-material/eventdetail/120/-/ccpbiosim-training-week-2019) at the University of Bristol, where he has received training in current computational methodologies and tools (Python, Jupyter Notebooks, BioSimSpace, Free energy calculations and etc.). Waris has also attended a number career and professional workshops and courses at Queen's University Belfast to improve his writing skills, to learn more about time management and leadership. Drs. Tikhonova and Singh have hosted and trained summer students (Tatiana Bezakova and Caitlin Duffy) supported by IAESTE and Nuffield Foundation in computer simulations of BSH enzymes (June-August, 2019). How have the results been disseminated to communities of interest?We have actively and effectively disseminated new information and procedures to scientists, producers, organizations, industries, government regulatory agency, and veterinarians. Following are some examples: Dr. Jun Lin (PI) gave invited talks focused on developing BSH inhibitor-based alternative to antibiotic growth promoters in two institutions, which include Washington State University (April 18, 2019) and USDA/Agriculture Research Service center in Beltsville, Maryland (December 10, 2019). Dr. Jun Lin gave keynote talk in the First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products held in Ningbo, China (November 16-18, 2019) Dr. Susan Joyce (Co-PI, Republic of Ireland) gave invited plenary talks to disseminate results to communities of interest, such as First Symposium on Host Microbe Interactions (April, 2019 University of Bristol), Irish Mass Spectrometry Society Annual Conference (May 2019, Dublin Ireland), COST European Union NutREDOx COST Actions group (October 2019, Lisbon, Portugal). Dr. Susan Joyce also gave poster presentations in conferences, such as Irish Mass Spectrometry Society Annual Conference (May 2019, Dublin Ireland), Cell Symposia on Exercise Metabolism (May, 2019) Dr. Irina Tikhonova (Co-PI) gave an invited talk on BSH structure and function at Trinity College Dublin (April 10, 2019). Dr. Warispreet Singh gave poster presentations at the QUB Postdoctoral symposium (March 22, 2019, Riddel Hall), at 7th Annual CCPBioSim Conference - Frontiers in Bimolecular Simulations (September 4 -6, 2019, Bristol). What do you plan to do during the next reporting period to accomplish the goals?To ensure seamless and efficient cooperation for this Tri-partite collaborative project, Dr. Irina Tikhonova (Co-PI, Northern Ireland) hosted a one day meeting on September 12, 2019 in School of Pharmacy at Queen's University Belfast. Participants include three key investigators (Lin, Joyce, and Tikhonova) and two postdocs (Dr. Waris Singh from Tikhonova's lab, and Dr. Sarah Long from Dr. Joyce's lab). Each participants gave 1-3 oral presentations. This is a highly productive meeting for summarizing previous collaborative work, for prioritizing projects, and for developing new innovative ideas to advance research in this significant area. Following are brief plan for the studies that will be performed during the next reporting period. Based on the results from dosimetric trials and our recent progress on developing encapsulation technology to improve bioavailability of BSH inhibitor in vivo, we plan to perform a large chicken trial with comprehensive nutrition, physiology, microbiome, and metabolome studies as proposed in Objective 1 and 2. For this trial, we will include a new treatment in which fed will be supplemented with encapsulated BSH inhibitor (Lin/Zhai, USA & Joyce, Ireland). Determining of presence of Taurine breakdown intermediates via. whole strain but plasmid borne BSH activity and their relevance in biological material, as well as purified BSHs representing the 5 clades identified above (Joyce/Lin). Purification of classified gut represented BSH proteins to delimit their interaction abilities and specific activity in vitro (Joyce/Lin) Analysis of bespoke, designed mutations, mutagenesis of targeted amino acids in L. acidophilus BSH for bile acid metabolism for further activity modeling and simulation (Tikhonova, Northern Ireland/Lin/Joyce) Molecular dynamics simulations of BSH isotypes to identify a conformational space to be used for design of pan-based BSH inhibitors (Tikhonova/Lin/Joyce) Purification of classified gut represented BSH proteins to delimit their interaction abilities and specific activity in vitro (Joyce/Lin) The virtual screening studies will be performed to identify a panel of promising new BSH inhibitors (Tikhonova) and the new inhibitors will be verified using standard BSH assay (Lin). We plan to generate three peer-reviewed journal articles resulting from this collaborative project in 2020.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate identified BSH inhibitors in a well-simulated chicken experimental system. Following completion of the proposed three large scale dosimetric pen trials, we have performed all data analyses to determine and compare various growth parameters among different treatment groups, such as body weight (BW), BW gain, feed intake, and feed conversion ratio. The experiment revealed optimal levels of BSH inhibitors for riboflavin (6.6-20 ppm), for CAPE (10-20 ppm) and for carnosic acid (10-20 ppm). However, statistical analysis did not show significant dose-dependent effects of BSH inhibitor across a wide dosage range (0 - 160 ppm) (Joyce/Lin, USA). Based on the findings from chicken trials, we speculated the bioavailability of the hydrophobic BSH inhibitors could be enhanced using cost effective encapsulation technology. Thus, we developed and optimized an encapsulation technology. Recent in vitro digestion experiment demonstrated that bioavailability of the encapsulated BSH inhibitor increased >30 fold when compared to unencapsulated BSH inhibitor. (Lin, USA) Objective 2. Understand the mechanistic basis of BSH inhibitors through analysis of ex vivo samples. To better link growth performance data to intestinal BSH activity, a total of 136 ileal content samples collected from individual chickens (Objective 1) were shipped to Ireland partner for bile profile analysis. Specifically, each ileal sample was processed and subjected to UPLC-MS analysis; a total of 32 different bile acid moieties were analyzed for each sample. BSH inhibitors Key indicators of inhibition were reductions in downstream bile acid processing so that secondary and free bile acids were reduced and conjugated bile acids were enriched. (Joyce, Ireland & Zhai/Lin, USA) Crystal structure indicated that deconjugation of Taurine conjugated bile acids may result in a metabolite of taurine, 2-amino-2-hydroxyethane-1-sulfonic acid (TAW), fixed to L. acidophilus BSH enzyme, a novel finding that may provide new insights into catalytic mechanism of BSH enzymes. In order to investigate if this was the case, the breakdown metabolites of Taurine by the microbiota were identified (9 in total). Methodology (Chaimbault et al. 2004; Mosier et al., 2013) was adapted and developed for liquid chromatography Mass Spectrometry to detect these taurine catabolism. Detection of the majority of these intermediates in biological material by application of control strains known to produce a subset of them (isethionic acid, sulfoacetylaldehyde, acetyl CoA) and through BSH carrying strains (L. acidophilus and plasmid bourne BSH). Further work is underway with bacterial co-incubations as cell extracts to determine if a consequence of BSH activity is the sequestering of TAW. (Joyce, Ireland & Lin, USA) Objective 3. Understand BSH structure-function relationships and structure-based design of BSH inhibitors. Based on in-depth structural analysis, a panel of amino acid residues were identified for playing a critical role in BSH enzymatic activity and substrate spectrum (Tikhonova, Northern Ireland). Site-directed amino acid substitution was performed to generate 10 desired BSH mutants (L20W, A58F, F130V, L134Q, A137S, F208A, R224A, N262A, N262T) (Lin, USA). These ten BSH mutants has been shipped to Dr. Joyce (Ireland) for comprehensive BSH activity assay in the near future. Mutational analysis was performed (Tikhonova, Northern Ireland) and bespoke mutations were created to alter residues deemed essential for L. acidophilus deconjugation ability. These mutations were subsequently analysed through in vitro co-incubation assays. Western blot assay indicated complete protein production (Joyce, Ireland). BSH has been cloned representing 23 different BSHs from gut bacteria according to the literature (Jones et al., 2008; Song et al., Liang et al., 2019). They were cloned into pET21b, induced under the same T7 promoter system and compared by sequence and BA deconjugation activity to represent just 5 different clades in the context of the gut. This work revealed large variation in BSH activity even in bacterial strains that share high BSH sequence homology and that the application of UPLC-MS provided substrate profiles so that they could be functionally classified into just 5 clades. This novel data which broadens previous knowledge of BSH activity and has provided information critical for understanding the relevance of BSH expression and function within the gut microbiome and in examining BSH inhibitor function (Joyce, Ireland). We have performed retrospective computational analysis of available BSH inhibitors obtained via HTS to identify suitable docking and scoring programs for the prospective compound screenings. We then conducted virtual screening of FDA, NPASS, Nutraceuticas databases against the BSH structures and identified the list of 300 highly-scored compounds. The selection of compound hits for the experimental test is underway. (Tikhonova, Northern Ireland and Lin, USA).
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Xu, F., X.J. Hu, W. Singh, W. Geng, I. Tikhonova, J. Lin. 2019. The complex structure of the bile salt hydrolase from Lactobacillus salivarius reveals the residues contributing to catalysis and substrate specificity. Scientific Reports. 9:12438. DOI: 10.1038/s41598-019-48850-6
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Sarah L. Long and Susan A. Joyce. 2019. Bile Acids as Receptor Ligands in Metabolic Processes-The FXR Connection. Journal of Food and Nutritional Sciences International http://scholarsinsight.org/journals/current-issues/24-Article.pdf
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Prete, R., Long S.L., , Lopez Gallardo, A., Gahan, C., Corsetti, A. & S.A. Joyce. 2020. Beneficial bile acid metabolism from Lactobacillus plantarum of food origin. Scientific Reports 10, 1165 (2020). https://doi.org/10.1038/s41598-020-58069-5
- Type:
Journal Articles
Status:
Under Review
Year Published:
2020
Citation:
Geng, W., S. Long, Y.J. Chang, A. Saxton, S. A. Joyce, J. Lin. 2019. Evaluation of in vivo efficacy of bile salt hydrolase inhibitors using chicken model system. Scientific Reports. (Resubmitted with minor revisions)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Lin, J. 2019. Sustainability and safety of animal agriculture: emerging threats and innovative solutions. The First Symposium on Plant/Animal Health and Quality/Safety of Agro-Products. Ningbo, China, November 16-18, 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Zhang, B., and W. Zhai. 2019. Effects of riboflavin on growth performance, processing yield, and internal organ development of Ross 708 male broilers. Poult. Sci. 98 (E-Suppl. 1). Page 27. Abstract # M84. 2019 International Poultry Scientific Forum in Atlanta, GA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Singh W, Joyce SA, Lin J, Tikhonova IG. 2019. Structure-based Search of Bile Salt Hydrolase Inhibitors. 7th Annual CCPBioSim Conference Frontiers in Bimolecular Simulations (September 4 -6, 2019, Bristol).
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2019
Citation:
Sarah L. Long. 2019. Ph.D. dissertation. APC Microbiome Ireland, School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland. Ph.D. Mentor: Dr. Susan A. Joyce.
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Progress 01/01/18 to 12/31/18
Outputs
Target Audience:Researchers, animal scientists, poultry producers, industry organizations, government officers, regulatory officers, students. Following is an outline of our major outreach plan: • SCIENTIFIC PEER-REVIEWED JOURNAL ARTICLES. • PRESENTATIONS AT SCIENTIFIC MEETINGS. • POSTDOC AND GRADUATE STUDENT TRAINING. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? USA. A Ph.D student was actively involved in all large chicken trials to perform dosimetric studies for the three BSH inhibitors. This graduate student received comprehensive training and obtained hands on experience on chicken nutrition, physiology, and development of BSH-based non-antibiotic feed additive. This project also partly supported a Postdoc to gain training in the areas of molecular microbiology, bioinformatics, and enzymology. Finally, this project supported Dr. Jun Lin (PI) to attend a workshop on priority-setting for antibiotic stewardship in animal agriculture in Washington DC in December, 2018. <!-- --> Republic of Ireland. This project provided Dr. Susan Joyce (Co-PI) opportunities to hire Postdoc and graduate student to perform basic and translational studies related to BSH enzymes. With aid of this project, Dr. Joyce and her trainees also have been involved in various professional development, which include but are not limited to the Health and safety training and Eigeum on line Research Integrity course. They have registered and will participate multiple workshop and professional meetings in 2019. Northern Ireland. A postdoc was hired and has been gaining experience in commercial software for structure-based drug design. He will attend the Computer-Aided Drug Design Workshop on March 14, 2019 in Cambridge, UK to increase his knowledge in small molecule virtual screening and protein modelling: https://chemcomp.com/Workshops-Europe.htm How have the results been disseminated to communities of interest?We have actively and effectively disseminated new information and procedures to scientists, producers, organizations, industries, government regulatory agency, and veterinarians. Following are some examples: Dr. Jun Lin (PI) gave invited talks focused on developing BSH inhibitor-based alternative to antibiotic growth promoters at multiple institutions, such as Case Western Reserve University (April 12, 2018), Qinghai University (China, June 26, 2018), Zhejiang Academy of Agricultural Sciences (China, July 12, 2018), Nanjing Agricultural University (China, July 17, 2018), and APC Microbiome Institute (Oct 3, 2018). Dr. Jun Lin lab personnel attended American Society for Microbiology Annual Meeting (June 7-11, Atlanta, GA) and the Focused Meeting 2018: Microbiomes Underpinning Agriculture.(October 1-2, Cork, Ireland) to present research findings for developing BSH inhibitor-based non-antibiotic feed additives. Dr. Jun Lin also attended workshop on priority-setting for antibiotic stewardship in animal agriculture, hosted by The Pew Charitable Trusts and the Foundation for Food and Agriculture Research (FFAR) in Washington DC in December, 2018. Dr. Lin advocated development of innovative science-based non-antibiotic feed additives, such as BSH inhibitors, to improve animal production, feed efficiency and sustainability. Dr. Susan Joyce (Co-PI, RoI) gave invited talks to disseminate results to communities of interest, such as The International Scientific ConferenceonProbiotics and Prebiotics (July 2018 Budapest), World Congress in Gastroenterology (Rome, Italy 2018), Third Prebiotic Conference China (October 2018), Wellcome- Cambridge- Microbiome (December 2018) Dr. Irina Tikhonova (Co-PI, NI) and PDRA have been organizing and will hold a one-day molecular modelling workshop on April 29, 2019 at QUB to demonstrate the use and application of computational protocols and scripts developing during the research project. Our workshop is targeted at an audience of computational and experimental chemists and biologists working in academia and industry. This will help to start dissemination of research findings to a large and diverse research community, increasing the potential economic and social impact of the research. What do you plan to do during the next reporting period to accomplish the goals? We will perform a large chicken trial with comprehensive nutrition, physiology, microbiome, and metabolome studies as proposed in Objective 1 and 2. We will analyze 23 BSHs functionally and in silico We will study BSH/TCA complex structure and get more insights into catalysis mechanism of BSH. Pharmacophore models will be developed using the known eight inhibitor molecules and these models will be used in conjunction with the docking programs to screen the databases of lead like compounds available from Zinc15 and FDA databases. The virtual screening studies will be performed on the lead like compounds available from Zinc15 and Natural and Natural-Like Compound Database. We plan to generate three peer-reviewed journal articles resulting from this collaborative project in 2019.
Impacts
What was accomplished under these goals?
Objective 1. Evaluate identified BSH inhibitors in a well-simulated chicken experimental system. Three large broiler pen trials have been completed to determine different doses of supplemented BSH inhibitors on chicken growth performance. The data from all three large chicken pen trials have been analyzed and will be used for design of the proposed comprehensive growth and nutrition measurements. Objective 2. Understand the mechanistic basis of BSH inhibitors through analysis of ex vivo samples. The collaborator Dr. Susan Joyce performed metabolomics analysis of chicken fecal samples to link growth performance parameters to gut bile acid profiles. BSH was characterized from a range of strains by bile co-incubation assay for broad substrate range. Antibodies were made against an internal BSH sequence and they were applied to examine expression levels. Levels of expression varied - although the same promoter is present.Substrate specificity and activity also varied. Objective 3. Understand BSH structure-function relationships and structure-based design of BSH inhibitors. A series of 23 BSH were selected for investigations based on their sequence and classication from previous publications. They are currently being examined in silico and for functional activity (Joyce, RoI) and will be examined for structure function (Tikhonova, NI) and they will be examined for interaction with specific BSH inhibitors (Lin, USA). Extensive structure-function studies have been performed on the Lactobacillus salivarius BSH under active and seamless collaborations among Lin (PI, USA), Tikhonova (Co-PI, NI), and Joyce (Co-PI, RoI). The Molecular Dynamics Simulations, QMMM and Molecular Docking studies were performed. Major findings include I) MD simulations were in accordance with the X-ray structure and provided a starting structure to perform QMMM analysis. II) The potential energy scan of the nucleophilic attack of the thiolate anion on the carbonyl carbon of the amide bond of the substrate (GCH) was computed using DFT using wB97XD-6-31G(d,p). The proton transfer from the thiol group to the N-terminal amine was performed using the similar methodology. It was found out that the ionic form was more stable than the neutral form, providing insights into catalysis mechanism of BSH. III) Homology model for the following isoforms of BSH were created using Modeller program: (I) F5VED8 (Lactobacillus salivarius NIAS840), (II) Q1WR93 (Lactobacillus salivarius strain UCC118), (III) C7AQX8 (Lactobacillus salivarius) and (IV) A0A076JMQ3 (Bifidobacterium adolescentis). The MD simulations were performed on all these models to understand the flexibility of loops surrounding the binding site Performed retrospective screening of the high-throughput screening(HTS)-compound library to find a protocol for a prospective virtual screening, a critical progress for structure-based design of BSH inhibitors. Major findings include I) The known eight inhibitor molecules were then docked into the binding site of BSH (PDB: 5Y7P) using Glide, Plants1.2, AutodockVina and Dock6.8 docking programs to understand the mode of inhibitor binding. Several binding modes have been identified. The validation of the binding sites requires mutagenesis or a crystallographic structure. II) The Glide with XP scoring function provides the best results in comparison to the other docking programs in terms of the enrichment score of the eight known binders in comparison to the non-binders. III) The virtual screening study was performed via docking of the FDA approved database using the X-ray structure of 5Y7P - the results in the process of analysis
Publications
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2018
Citation:
Wenjing Geng, 2018. Bile salt hydrolase: from basic science to translational innovation. Ph.D. Dissertation
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Geng, W., C. Gahan, S. Joyce, J. Chang, A. Saxton, J. Lin. 2018. In vivo evaluation of bile salt hydrolase inhibitors using broiler chicken. American Society for Microbiology Annual Meeting. June 7-11, Atlanta, GA
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Lin, J., W. Geng, S. Long, S. Joyce. 2018. Improved animal husbandry through inhibition of gut microbial bile salt hydrolase. Focused Meeting 2018: Microbiomes Underpinning Agriculture. October 1-2, Cork, Ireland
- Type:
Journal Articles
Status:
Under Review
Year Published:
2019
Citation:
Xu, F., W. Geng, X.J. Hu, W. Singh, I. Tikhonova, S. Joyce, J. Lin. 2018. The complex structure of the bile salt hydrolase from Lactobacillus salivarius reveals the residues contributing to catalysis and substrate specificity. Scientific Reports
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