2021 Fiscal Year Research-status Report
Metabolomics-driven approach to understand antibiotics induction ability mechanism in Streptomyces coelicolor
Project/Area Number |
20K05787
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Research Institution | Osaka University |
Principal Investigator |
PUTRI SASTIA 大阪大学, 工学研究科, 助教 (50729796)
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Project Period (FY) |
2020-04-01 – 2023-03-31
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Keywords | Streptomyces coelicolor / antibiotics production / gamma-butyrolactones |
Outline of Annual Research Achievements |
Previously, the correlation between cAMP levels and antibiotics production was identified. Furthermore, cAMP supplementation to S. coelicolor culture led to improvements of cell growth and antibiotic production as well as an increase of guanine levels and the expression level of purine metabolism genes. The results have been published in two scientific articles. As a follow up study, other factors that affect antibiotics production was investigated. In this fiscal year, comparative metabolite analysis was conducted to investigate the role of gamma butyrolactone (GBL)signaling molecule and its relationship to antibiotics production. Among the GBL, SCB1 acts in a highly specific manner to elicit the production of actinorhodin (ACT, blue color) and undecylprodigiosin (RED, red color) (Takano et al., 2000). The gene mutations led to either overproduction or delay of antibiotic production. Hence, the objective of the study is to analyze the changes in primary metabolite levels caused by gene deletions in the SCB1-ScbR system genes in Streptomyces coelicolor. Prior to performing comparative metabolome analyses, growth curve and phenotypic checks were performed. This study is still ongoing and will be continued in the next fiscal year.
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Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
The objective of this project is to identify metabolites that strongly correlate with antibiotics production. This correlation is then used to devise a strategy to improve antibiotics production. The project objective has been achieved in the first year of this project and the results have been published in two reputable journals. In the remaining two years, a follow up study to investigate other factors influencing antibiotics production is studied. In this follow up study, the focus is to study about the role of gamma butyrolactones in modulating antibiotics production. Among the GBL, SCB1 acts in a highly specific manner to elicit the production of actinorhodin (ACT, blue color) and undecylprodigiosin (RED, red color) (Takano et al., 2000). The SCB signaling system is the focus of the remaining project.
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Strategy for Future Research Activity |
The purpose of the study is to analyze the changes in primary metabolite levels caused by gene deletions in the SCB1-ScbR system genes in Streptomyces coelicolor. The expected outcomes of this study are: 1.The comparison of metabolic profiles of the four S. coelicolor strains: (1) Streptomyces coelicolor A3(2) M145 wildtype, (2) M751 (M145 with scbA knockout), (3) M752 (M145 with scbR knockout), (4) LW94 (M145 (scbA-scbR)knockouts + pTE1062) 2.Discovery of significant metabolite changes in knockout strains (M751, M752, LW94) compared to the wildtype strain The study is outlined to, first, screen for trends of metabolite changes upon GBL and antibiotic production through exhaustive metabolite profiling using LC/MS/MS and GC/MS/MS, then determine the specific metabolites that differ in each mutant, and lastly, do data analysis to try find the key metabolites in relation to each strains’ activity. Due to the pandemic situation, construction of new strains by our collaborator in the University of Manchester is postponed due to prolonged lockdown in the UK last year. In the final year of this project, new insights involving the correlation between GBL and antibiotics production is expected to shed a new light on how to modulate antibiotics production in Streptomyces coelicolor.
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Causes of Carryover |
Due to the pandemic situation, travel could not be conducted in 2021 fiscal year and therefore will be done in 2022 fiscal year.
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Research Products
(1 results)