Elucidating Physiology of Complex Microbial Systems through Co-Culture 13c-Metabolic Flux Analysis
Metabolic Engineering Conference
2016
Metabolic Engineering 11
Poster Session
Poster Session 3
Tuesday, June 28, 2016 - 5:30pm to 7:00pm
Nikodimos Gebreselassie
Nikodimos Gebreselassie
3
3968
2016-02-01T21:44:00Z
2016-02-01T21:47:00Z
1
479
2735
22
6
3208
15.00
Clean
Clean
false
false
false
false
EN-US
X-NONE
X-NONE
DefSemiHidden="false" DefQFormat="false" DefPriority="99"
LatentStyleCount="371">
UnhideWhenUsed="true" QFormat="true" Name="heading 2"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 3"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 4"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 5"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 6"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 7"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 8"/>
UnhideWhenUsed="true" QFormat="true" Name="heading 9"/>
Name="index 1"/>
Name="index 2"/>
Name="index 3"/>
Name="index 4"/>
Name="index 5"/>
Name="index 6"/>
Name="index 7"/>
Name="index 8"/>
Name="index 9"/>
UnhideWhenUsed="true" Name="toc 1"/>
UnhideWhenUsed="true" Name="toc 2"/>
UnhideWhenUsed="true" Name="toc 3"/>
UnhideWhenUsed="true" Name="toc 4"/>
UnhideWhenUsed="true" Name="toc 5"/>
UnhideWhenUsed="true" Name="toc 6"/>
UnhideWhenUsed="true" Name="toc 7"/>
UnhideWhenUsed="true" Name="toc 8"/>
UnhideWhenUsed="true" Name="toc 9"/>
Name="Normal Indent"/>
Name="footnote text"/>
Name="annotation text"/>
Name="header"/>
Name="footer"/>
Name="index heading"/>
UnhideWhenUsed="true" QFormat="true" Name="caption"/>
Name="table of figures"/>
Name="envelope address"/>
Name="envelope return"/>
Name="footnote reference"/>
Name="annotation reference"/>
Name="line number"/>
Name="page number"/>
Name="endnote reference"/>
Name="endnote text"/>
Name="table of authorities"/>
Name="macro"/>
Name="toa heading"/>
Name="List"/>
Name="List Bullet"/>
Name="List Number"/>
Name="List 2"/>
Name="List 3"/>
Name="List 4"/>
Name="List 5"/>
Name="List Bullet 2"/>
Name="List Bullet 3"/>
Name="List Bullet 4"/>
Name="List Bullet 5"/>
Name="List Number 2"/>
Name="List Number 3"/>
Name="List Number 4"/>
Name="List Number 5"/>
Name="Closing"/>
Name="Signature"/>
UnhideWhenUsed="true" Name="Default Paragraph Font"/>
Name="Body Text"/>
Name="Body Text Indent"/>
Name="List Continue"/>
Name="List Continue 2"/>
Name="List Continue 3"/>
Name="List Continue 4"/>
Name="List Continue 5"/>
Name="Message Header"/>
Name="Salutation"/>
Name="Date"/>
Name="Body Text First Indent"/>
Name="Body Text First Indent 2"/>
Name="Note Heading"/>
Name="Body Text 2"/>
Name="Body Text 3"/>
Name="Body Text Indent 2"/>
Name="Body Text Indent 3"/>
Name="Block Text"/>
Name="Hyperlink"/>
Name="FollowedHyperlink"/>
Name="Document Map"/>
Name="Plain Text"/>
Name="E-mail Signature"/>
Name="HTML Top of Form"/>
Name="HTML Bottom of Form"/>
Name="Normal (Web)"/>
Name="HTML Acronym"/>
Name="HTML Address"/>
Name="HTML Cite"/>
Name="HTML Code"/>
Name="HTML Definition"/>
Name="HTML Keyboard"/>
Name="HTML Preformatted"/>
Name="HTML Sample"/>
Name="HTML Typewriter"/>
Name="HTML Variable"/>
Name="Normal Table"/>
Name="annotation subject"/>
Name="No List"/>
Name="Outline List 1"/>
Name="Outline List 2"/>
Name="Outline List 3"/>
Name="Table Simple 1"/>
Name="Table Simple 2"/>
Name="Table Simple 3"/>
Name="Table Classic 1"/>
Name="Table Classic 2"/>
Name="Table Classic 3"/>
Name="Table Classic 4"/>
Name="Table Colorful 1"/>
Name="Table Colorful 2"/>
Name="Table Colorful 3"/>
Name="Table Columns 1"/>
Name="Table Columns 2"/>
Name="Table Columns 3"/>
Name="Table Columns 4"/>
Name="Table Columns 5"/>
Name="Table Grid 1"/>
Name="Table Grid 2"/>
Name="Table Grid 3"/>
Name="Table Grid 4"/>
Name="Table Grid 5"/>
Name="Table Grid 6"/>
Name="Table Grid 7"/>
Name="Table Grid 8"/>
Name="Table List 1"/>
Name="Table List 2"/>
Name="Table List 3"/>
Name="Table List 4"/>
Name="Table List 5"/>
Name="Table List 6"/>
Name="Table List 7"/>
Name="Table List 8"/>
Name="Table 3D effects 1"/>
Name="Table 3D effects 2"/>
Name="Table 3D effects 3"/>
Name="Table Contemporary"/>
Name="Table Elegant"/>
Name="Table Professional"/>
Name="Table Subtle 1"/>
Name="Table Subtle 2"/>
Name="Table Web 1"/>
Name="Table Web 2"/>
Name="Table Web 3"/>
Name="Balloon Text"/>
Name="Table Theme"/>
Name="List Paragraph"/>
Name="Intense Quote"/>
Name="Subtle Emphasis"/>
Name="Intense Emphasis"/>
Name="Subtle Reference"/>
Name="Intense Reference"/>
UnhideWhenUsed="true" Name="Bibliography"/>
UnhideWhenUsed="true" QFormat="true" Name="TOC Heading"/>
Name="Grid Table 1 Light Accent 1"/>
Name="Grid Table 6 Colorful Accent 1"/>
Name="Grid Table 7 Colorful Accent 1"/>
Name="Grid Table 1 Light Accent 2"/>
Name="Grid Table 6 Colorful Accent 2"/>
Name="Grid Table 7 Colorful Accent 2"/>
Name="Grid Table 1 Light Accent 3"/>
Name="Grid Table 6 Colorful Accent 3"/>
Name="Grid Table 7 Colorful Accent 3"/>
Name="Grid Table 1 Light Accent 4"/>
Name="Grid Table 6 Colorful Accent 4"/>
Name="Grid Table 7 Colorful Accent 4"/>
Name="Grid Table 1 Light Accent 5"/>
Name="Grid Table 6 Colorful Accent 5"/>
Name="Grid Table 7 Colorful Accent 5"/>
Name="Grid Table 1 Light Accent 6"/>
Name="Grid Table 6 Colorful Accent 6"/>
Name="Grid Table 7 Colorful Accent 6"/>
Name="List Table 1 Light Accent 1"/>
Name="List Table 6 Colorful Accent 1"/>
Name="List Table 7 Colorful Accent 1"/>
Name="List Table 1 Light Accent 2"/>
Name="List Table 6 Colorful Accent 2"/>
Name="List Table 7 Colorful Accent 2"/>
Name="List Table 1 Light Accent 3"/>
Name="List Table 6 Colorful Accent 3"/>
Name="List Table 7 Colorful Accent 3"/>
Name="List Table 1 Light Accent 4"/>
Name="List Table 6 Colorful Accent 4"/>
Name="List Table 7 Colorful Accent 4"/>
Name="List Table 1 Light Accent 5"/>
Name="List Table 6 Colorful Accent 5"/>
Name="List Table 7 Colorful Accent 5"/>
Name="List Table 1 Light Accent 6"/>
Name="List Table 6 Colorful Accent 6"/>
Name="List Table 7 Colorful Accent 6"/>
communities carry out efficient biological transformations that are the result
of multiple complementary metabolic systems working together. In bioprocess
applications, co-culture systems have unique advantages over mono-culture
systems in optimizing substrate utilization and product yield, i.e. co-culture systems
allow for the independent optimization and assembly of various complementary
metabolic capabilities into functional systems, where the diversity of
metabolic pathways and ability of microorganisms to exchange metabolites
dramatically increases the space of possible metabolic conversions. normal">Co-culture
systems are now increasingly applied in metabolic engineering to optimize utilization
of mixed substrates, for production of mixed products, and distribution of
complex metabolic pathways among species. Among the many omics tools used to
gain insight into the physiology of these systems, fluxomics
is the most direct and relevant method to study the normal">in vivo metabolic state of co-cultures. In the past, 13C-metabolic
flux analysis (13C-MFA) of co-culture systems required physical
separation of proteins or cells to resolve individual population fluxes. In
this work, we demonstrate for the first time that it is possible to determine
metabolic flux distributions in multiple species simultaneously without the
need for physical separation of cells or proteins, or overexpression of
species-specific products. Instead, metabolic fluxes for each species in a
co-culture are estimated directly from isotopic labeling of total biomass
obtained using conventional mass spectrometry approaches such as GC-MS. In
addition to determining metabolic fluxes, the approach quantifies the relative
population size of each species in a mixed culture and interspecies metabolic
exchange, thus enabling detailed studies of species dynamics and interactions
in co-cultures. normal">We
will demonstrate application of our novel co-culture 13C-MFA approach
on the following three diverse and industrially relevant co-culture systems: margin-bottom:0in;margin-left:27.0pt;margin-bottom:.0001pt;mso-add-space:auto;
text-indent:-.25in;line-height:normal;mso-list:l1 level1 lfo2"> "Times New Roman"'>i.
mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi'>A
co-culture system in which each species consumes the same substrate but
metabolizes the substrate through different pathways, specifically a co-culture
of E. coli knockouts Δpgi and Δzwf. margin-bottom:0in;margin-left:27.0pt;margin-bottom:.0001pt;mso-add-space:auto;
text-indent:-.25in;line-height:normal;mso-list:l1 level1 lfo2"> "Times New Roman"'>ii.
mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi'>A
co-culture system in which each species consumes the same substrate but
produces a different product, specifically a co-culture of wild-type E. coli and normal"> S. cerevisiae. margin-bottom:0in;margin-left:27.0pt;margin-bottom:.0001pt;mso-add-space:auto;
text-indent:-.25in;line-height:normal;mso-list:l1 level1 lfo2"> "Times New Roman"'>iii.
mso-bidi-font-family:"Times New Roman";mso-bidi-theme-font:minor-bidi'>A
co-culture system in which one species consumes a particular substrate and produces
a by-product that serves as the substrate for the second species, specifically a
glucose consuming E. coli and a
glucose non-consuming E. coli strain
(ΔptsI) that uses acetate produced by the first
strain as its carbon source. normal"> mso-fareast-font-family:"Times New Roman"'>Through the use of co-culture 13C-MFA,
we are able to elucidate the metabolic state of the various co-culture systems
as well as investigate species dynamics and interspecies metabolite exchange. Moreover,
an improved metabolic network model of S.
cerevisiae for 13C-MFA was developed through parallel tracer
experiments. In summary, our work provides a powerful novel approach to study
co-culture systems that will greatly facilitate rational design of co-culture systems
in metabolic engineering. normal">