Characterization of Characterization of HTCC5015: A Novel HTCC5015: A Novel Bacterial Isolate from Bacterial Isolate from the Sargasso Sea the Sargasso Sea By: Marie Johnson By: Marie Johnson Dr. Stephen Giovannoni Dr. Stephen Giovannoni Laboratory of Marine Laboratory of Marine Microbiology Microbiology
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Characterization of HTCC5015: A Novel Bacterial Isolate from the Sargasso Sea By: Marie Johnson Dr. Stephen Giovannoni Laboratory of Marine Microbiology.
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Characterization of HTCC5015: A Characterization of HTCC5015: A Novel Bacterial Isolate from the Novel Bacterial Isolate from the
Sargasso SeaSargasso Sea
By: Marie JohnsonBy: Marie JohnsonDr. Stephen Giovannoni Laboratory of Dr. Stephen Giovannoni Laboratory of
Marine MicrobiologyMarine Microbiology
BackgroundBackground
• Alternative sources of energy / clean the Alternative sources of energy / clean the environment environment
• Isolated by Dr. Uli Stingl from the Sargasso SeaIsolated by Dr. Uli Stingl from the Sargasso Sea
• Maximum density in culture is between 10Maximum density in culture is between 1055 and10 and106 6
cells/mL cells/mL
BackgroundBackground
• No additional growth when inoculated into seawater No additional growth when inoculated into seawater containing 90 different carbon compoundscontaining 90 different carbon compounds
• Oregon vs. Bermuda coastal watersOregon vs. Bermuda coastal waters
• Analysis of 16s rRNA – related to thiotrophic Analysis of 16s rRNA – related to thiotrophic symbionts symbionts
Questions to ExploreQuestions to Explore• Which compound(s) does HTCC5015 use as a carbon Which compound(s) does HTCC5015 use as a carbon
source?source?
• Are HTCC5015’s energy needs similar to that of Are HTCC5015’s energy needs similar to that of thiotrophic symbionts?thiotrophic symbionts?
StrategyStrategy
• Determine HTCC5015’s nutrient requirements Determine HTCC5015’s nutrient requirements therefore achieving a higher maximum cell therefore achieving a higher maximum cell density in culture. density in culture.
Growth Experiment 1Growth Experiment 1
• Test three separate compounds + control Test three separate compounds + control (LNHM)(LNHM)
– DMSP DMSP – Glycine BetaineGlycine Betaine
– CholineCholine
Sulfur compounds possibly used in Sulfur compounds possibly used in metabolic processesmetabolic processes
• Test two compounds separately + control (sea Test two compounds separately + control (sea water)water)
– LNHM LNHM
– Potassium Nitrate Potassium Nitrate
Base media containing filtered sea water and fortified with Base media containing filtered sea water and fortified with vitamins, iron, phosphorus, and nitrogen – in the form of NHvitamins, iron, phosphorus, and nitrogen – in the form of NH33
Experimental source of nitrogen in Experimental source of nitrogen in replacement of ammoniareplacement of ammonia
• Re-test sulfur compounds from experiment 4Re-test sulfur compounds from experiment 4– Find support to either accept or reject the previous results Find support to either accept or reject the previous results
that providing cystine may allow for an increased that providing cystine may allow for an increased maximum density in culturemaximum density in culture
• Analyze HTCC5015 genome to determine incomplete Analyze HTCC5015 genome to determine incomplete metabolic pathwaysmetabolic pathways– Formulate experiments with compounds unable to be Formulate experiments with compounds unable to be
made by HTCC5015 made by HTCC5015
AcknowledgementsAcknowledgements
• Howard Hughes Medical InstituteHoward Hughes Medical Institute• Kevin AhernKevin Ahern• Stephen Giovannoni Stephen Giovannoni • Kevin VerginKevin Vergin• Jim TrippJim Tripp• Tom RichardsonTom Richardson• Joshua KitnerJoshua Kitner• Uli StinglUli Stingl