The document discusses developing a bacterial co-culture system for consolidated bioprocessing of cellulose to biofuels. It begins by outlining the goals and challenges of consolidated bioprocessing. Initial experiments characterized growth of Cytophaga hutchinsonii and E. coli in a naive co-culture on cellulose. Dynamic modeling was used to design genetic circuits to control population dynamics. Parts for engineering C. hutchinsonii were developed, including a modular origin of replication. Overall progress was made in characterizing co-cultures, designing population control systems, and implementing genetic parts for consolidated bioprocessing.
The document analyzes the music video for the song "Live While We're Young" by One Direction. It discusses various filming techniques used in the video, including mid shots of the band playing guitar around a campfire, close ups of the members lip syncing to directly address the audience, and long shots showing the group together to emphasize their friendship. Quick cuts are used during a football scene to portray their fast-paced, youthful lifestyle. Costumes, lighting, settings and props are meant to portray the band and situation as fun, casual, and inviting for their target teenage audience.
A Risky Tomorrow beyond Traditional Human Resource Risk Management V1Matthew Bennett
This document provides an overview of traditional human resource risk management and introduces the concept of human risk management. It discusses how integrating psychological factors can help assess risks in key human resource areas like recruitment, health and safety, and change management. The document outlines the scope, objectives, and organizational impacts of exploring human risk management and how it may allow for better analysis of areas like human reactions, risk mitigation, and unpredictability. Finally, it provides background on traditional human resource management and risk management approaches.
El documento resume varias obras de arte, literatura y música relacionadas con la leyenda bíblica de Judit. Entre las obras de arte se encuentran pinturas de Botticelli, Miguel Ángel y Klimt, así como una escultura de Donatello, que representan el momento en que Judit decapita a Holofernes o muestra su cabeza. También se mencionan una obra de teatro y un soneto de Lope de Vega sobre el tema, así como varias obras musicales como oratorios y óperas.
Este documento resume los principales dioses del panteón griego. Zeus es el dios supremo que gobierna el cielo, aunque es infiel a su esposa Hera. Hestia es la diosa virgen del hogar y el fuego. Poseidón es el dios del mar y las tormentas que pelea con Atenea por ser el patrón de Atenas. Hades es el dios del inframundo que rapta a Perséfone por seis meses del año.
The document analyzes the music video for the song "Live While We're Young" by One Direction. It discusses various filming techniques used in the video, including mid shots of the band playing guitar around a campfire, close ups of the members lip syncing to directly address the audience, and long shots showing the group together to emphasize their friendship. Quick cuts are used during a football scene to portray their fast-paced, youthful lifestyle. Costumes, lighting, settings and props are meant to portray the band and situation as fun, casual, and inviting for their target teenage audience.
A Risky Tomorrow beyond Traditional Human Resource Risk Management V1Matthew Bennett
This document provides an overview of traditional human resource risk management and introduces the concept of human risk management. It discusses how integrating psychological factors can help assess risks in key human resource areas like recruitment, health and safety, and change management. The document outlines the scope, objectives, and organizational impacts of exploring human risk management and how it may allow for better analysis of areas like human reactions, risk mitigation, and unpredictability. Finally, it provides background on traditional human resource management and risk management approaches.
El documento resume varias obras de arte, literatura y música relacionadas con la leyenda bíblica de Judit. Entre las obras de arte se encuentran pinturas de Botticelli, Miguel Ángel y Klimt, así como una escultura de Donatello, que representan el momento en que Judit decapita a Holofernes o muestra su cabeza. También se mencionan una obra de teatro y un soneto de Lope de Vega sobre el tema, así como varias obras musicales como oratorios y óperas.
Este documento resume los principales dioses del panteón griego. Zeus es el dios supremo que gobierna el cielo, aunque es infiel a su esposa Hera. Hestia es la diosa virgen del hogar y el fuego. Poseidón es el dios del mar y las tormentas que pelea con Atenea por ser el patrón de Atenas. Hades es el dios del inframundo que rapta a Perséfone por seis meses del año.
El documento resume las biografías y logros científicos de cuatro importantes astrónomos: Nicolás Copérnico descubrió que la Tierra gira alrededor del Sol y no al revés; Galileo Galilei mejoró el telescopio y realizó observaciones astronómicas; Isaac Newton formuló las leyes del movimiento y de la gravedad universal; y Johannes Kepler formuló las tres leyes del movimiento planetario.
Chacón Producciones es una empresa de desarrollo de software fundada por Cesar Chacón que busca proveer soporte y mantenimiento de sistemas de información a nivel mundial para facilitar los estilos de vida. Su visión es convertirse en una gran empresa de desarrollo de software a nivel global que compita con Google y Apple en 10 años. Su misión es brindar tecnología actualizada a empresas y personas, ofreciendo soporte para computadoras y equipos informáticos. Sus metas incluyen establecer mayores índices de ventas de productos informá
Este documento presenta a cuatro científicos importantes de la edad moderna: Copérnico, que propuso el modelo heliocéntrico del sistema solar; Kepler, que elaboró una tesis geométrica explicando las distancias entre las órbitas planetarias; Galileo, que descubrió las leyes de la caída de los cuerpos y apoyó el modelo heliocéntrico; y Newton, que desarrolló el cálculo y demostró que las leyes de la mecánica celeste y terrestre son las mismas
For the IB Biology course. To get the editable pptx file, please make a donation to one of my chosen charities. More information: http://sciencevideos.wordpress.com/about/biology4good/
The comprehensive understanding of IslamFawad Kiyani
This document compares the ideologies of Islam and different systems of government. It discusses Islam as a religion and contrasts autocratic and democratic systems of government. However, most of the content is redacted and there are no other details provided about the key points or comparisons being made between these topics.
This document outlines the process for laboratory generation of monoclonal antibodies. It discusses strategies such as fed-batch culture in bioreactors and downstream purification techniques including protein A affinity chromatography, ion exchange chromatography, diafiltration, and gel filtration. It also provides an economic assessment of the process and discusses various scenarios and optimizations that could improve yield and reduce costs, such as using membrane chromatography or exploring mixed mode resins for purification.
The document outlines the course content for a fermentation technology class, including 10 chapters that cover topics like introduction to fermentation, fermentation processes and techniques, microbial rates, stoichiometry of microbial growth, heat and mass transfer in fermentation, and bioreactors. It provides examples of important fermentation products like ethanol, lactic acid, and antibiotics. It also includes diagrams of fermenter designs and considerations for fermentation medium composition and inoculation. The document serves as an overview of topics that will be covered in the class and provides background information on key concepts in fermentation technology.
The document summarizes experiments analyzing the transformation of chlorinated ethenes (TCE, cis-DCE, VC) by the Mycobacterium ELW-1 organism. Batch reactor experiments were conducted with varying initial concentrations of chlorinated ethenes and isobutene to determine maximum transformation rates. Results showed ELW-1 effectively transformed cis-DCE and VC but was uncertain for TCE bioremediation. Batch-grown cultures also transformed contaminants faster than chemostat-grown cultures. Future work will analyze ELW-1's ability to transform 1,4-dioxane.
This document discusses the use of micro-flow imaging (MFI) to analyze cell aggregation in thawed hematopoietic stem cell products. MFI allows visualization and precise quantification of cell clumping following cryopreservation. The study found that cell aggregates occur more often in thawed peripheral blood progenitor cell (PBPC) products compared to umbilical cord blood units. Treatment with DNase I after thawing can help disaggregate cells and increase the number of single cells available for transplantation. However, the impact of cell aggregation on clinical transplant outcomes remains unknown.
1. The document discusses the topics of fermentation technology and processes. It covers definitions of fermentation, important fermentation products, fermenter types, medium composition, inoculation, and microbial rates and stoichiometry.
2. Key aspects of fermentation covered include the use of stirred tank bioreactors, types of fermenters including batch, fed-batch and continuous reactors, and important industrial fermentation products like ethanol, lactic acid, and antibiotics.
3. Microbial rates are quantified as specific rates and yields, and stoichiometric equations are used to represent microbial growth coupling catabolism and anabolism based on substrate utilization and energy generation.
Fermentation is a form of metabolism where microorganisms break down organic compounds without oxygen. This document outlines the key topics in fermentation technology including: fermentation processes, microbial rates, stoichiometry of microbial growth and product formation, and important fermentation products. It also provides examples of stoichiometric calculations for microbial growth including determining yields, degrees of reduction, and setting up general stoichiometric equations using conservation principles.
The document describes the development of a high throughput platform for screening glycoside hydrolase enzymes based on oxime-NIMS. Key points:
1) JBEI developed an approach integrating oxime chemistries, acoustic printing and nanostructure-initiator mass spectrometry (Oxime-NIMS) to quantitatively analyze glycan products of GH reactions, including with complex biomass substrates.
2) They defined a standard panel of 12 substrates spanning plant cell wall linkages.
3) The platform was automated and tested by studying cellulases and their synergistic combinations across reaction conditions - generating datasets to enable enzyme cocktail design.
The document describes the principles and design of activated sludge wastewater treatment. Activated sludge processes use aerobic bacteria to consume organic matter in wastewater. Wastewater flows into an aeration tank where bacteria grow as they consume organic matter, then flows into a clarifier where bacteria settle as sludge. Sludge is partially recycled to the aeration tank to maintain bacteria populations. The design of activated sludge systems depends on parameters like hydraulic retention time, solids retention time, food to microorganism ratio, and dissolved oxygen levels. Typical designs include conventional, complete mix, extended aeration, and high rate aeration configurations.
JBEI Research Highlights December 2016 Irina Silva
The document describes research that engineered the 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in the yeast Saccharomyces cerevisiae. Key findings include:
- An engineered strain was able to grow in the absence of mevalonate, reaching 80% of growth compared to strains with mevalonate added.
- The DXP pathway became functional enough to support moderate growth without the mevalonate pathway, but growth on agar without mevalonate was not achieved.
- This work established a foundation for further optimizing yeast to produce bioenergy-relevant isoprenoids through the DXP pathway under industrial conditions.
JBEI Research Highlights - October 2021SaraHarmon4
This document summarizes a study that evaluated the potential for nutrient recovery from wet organic waste processing facilities in California to offset synthetic fertilizer demand. The study found that recovering nitrogen and phosphorus from organic waste streams through anaerobic digestion and separation techniques could meet 11% of the state's nitrogen and 29% of phosphorus fertilizer needs. Recovered nutrients would be in the form of liquid fertilizer, struvite, and compost. The approach provides a foundation for analyzing national-level nutrient flows and recovery potentials from bioenergy production.
Synthetic Biology for Plant ScientistsSachin Rawat
Tools of synthetic biology can be utilised to engineer metabolic pathways to optimize production of secondary metabolites and ligno-cellulose. The presentation describes an approach to develop an artificial positive feedback loop to increase accumulation of cell wall polysaccharides. These will decrease the cost of production of plant-based biofuels, paper and other plant products.
Cell Line Development: Reducing timelines and increasing titres fujifilmdiosynth
Cell line development: Reducing timelines and increasing titres by identification of host cell lines with improved characteristics. To develop a mammalian expression platform which rapidly leads to efficient, robust and high quality biomanufacturing processes
Scalability of Cell Culture Processes in Single-use Bioreactors using Differe...KBI Biopharma
Niket Bubna, Cameron T. Phillips, Sigma S. Mostafa and AbhinavA. Shukla. KBI Biopharma, Durham, NC
253rd ACS National Meeting & Exposition
April 2-6, 2017 • San Francisco, CA
#acsSanFran • www.acs.org/SanFran2017
- The study investigated using arbuscular mycorrhizal fungi (AMF) colonization and tree-herb interactions to enhance growth of legume trees for biofuel production on lead contaminated soil.
- AMF inoculation of legume trees increased overall biomass yield compared to non-inoculated plants under lead stress, likely by retaining lead in underground parts and improving nutrient acquisition.
- Combining AMF with planting legume trees with legume or grass herbs further optimized the system for phytoremediation of contaminated marginal lands and production of biofuel feedstocks.
El documento resume las biografías y logros científicos de cuatro importantes astrónomos: Nicolás Copérnico descubrió que la Tierra gira alrededor del Sol y no al revés; Galileo Galilei mejoró el telescopio y realizó observaciones astronómicas; Isaac Newton formuló las leyes del movimiento y de la gravedad universal; y Johannes Kepler formuló las tres leyes del movimiento planetario.
Chacón Producciones es una empresa de desarrollo de software fundada por Cesar Chacón que busca proveer soporte y mantenimiento de sistemas de información a nivel mundial para facilitar los estilos de vida. Su visión es convertirse en una gran empresa de desarrollo de software a nivel global que compita con Google y Apple en 10 años. Su misión es brindar tecnología actualizada a empresas y personas, ofreciendo soporte para computadoras y equipos informáticos. Sus metas incluyen establecer mayores índices de ventas de productos informá
Este documento presenta a cuatro científicos importantes de la edad moderna: Copérnico, que propuso el modelo heliocéntrico del sistema solar; Kepler, que elaboró una tesis geométrica explicando las distancias entre las órbitas planetarias; Galileo, que descubrió las leyes de la caída de los cuerpos y apoyó el modelo heliocéntrico; y Newton, que desarrolló el cálculo y demostró que las leyes de la mecánica celeste y terrestre son las mismas
For the IB Biology course. To get the editable pptx file, please make a donation to one of my chosen charities. More information: http://sciencevideos.wordpress.com/about/biology4good/
The comprehensive understanding of IslamFawad Kiyani
This document compares the ideologies of Islam and different systems of government. It discusses Islam as a religion and contrasts autocratic and democratic systems of government. However, most of the content is redacted and there are no other details provided about the key points or comparisons being made between these topics.
This document outlines the process for laboratory generation of monoclonal antibodies. It discusses strategies such as fed-batch culture in bioreactors and downstream purification techniques including protein A affinity chromatography, ion exchange chromatography, diafiltration, and gel filtration. It also provides an economic assessment of the process and discusses various scenarios and optimizations that could improve yield and reduce costs, such as using membrane chromatography or exploring mixed mode resins for purification.
The document outlines the course content for a fermentation technology class, including 10 chapters that cover topics like introduction to fermentation, fermentation processes and techniques, microbial rates, stoichiometry of microbial growth, heat and mass transfer in fermentation, and bioreactors. It provides examples of important fermentation products like ethanol, lactic acid, and antibiotics. It also includes diagrams of fermenter designs and considerations for fermentation medium composition and inoculation. The document serves as an overview of topics that will be covered in the class and provides background information on key concepts in fermentation technology.
The document summarizes experiments analyzing the transformation of chlorinated ethenes (TCE, cis-DCE, VC) by the Mycobacterium ELW-1 organism. Batch reactor experiments were conducted with varying initial concentrations of chlorinated ethenes and isobutene to determine maximum transformation rates. Results showed ELW-1 effectively transformed cis-DCE and VC but was uncertain for TCE bioremediation. Batch-grown cultures also transformed contaminants faster than chemostat-grown cultures. Future work will analyze ELW-1's ability to transform 1,4-dioxane.
This document discusses the use of micro-flow imaging (MFI) to analyze cell aggregation in thawed hematopoietic stem cell products. MFI allows visualization and precise quantification of cell clumping following cryopreservation. The study found that cell aggregates occur more often in thawed peripheral blood progenitor cell (PBPC) products compared to umbilical cord blood units. Treatment with DNase I after thawing can help disaggregate cells and increase the number of single cells available for transplantation. However, the impact of cell aggregation on clinical transplant outcomes remains unknown.
1. The document discusses the topics of fermentation technology and processes. It covers definitions of fermentation, important fermentation products, fermenter types, medium composition, inoculation, and microbial rates and stoichiometry.
2. Key aspects of fermentation covered include the use of stirred tank bioreactors, types of fermenters including batch, fed-batch and continuous reactors, and important industrial fermentation products like ethanol, lactic acid, and antibiotics.
3. Microbial rates are quantified as specific rates and yields, and stoichiometric equations are used to represent microbial growth coupling catabolism and anabolism based on substrate utilization and energy generation.
Fermentation is a form of metabolism where microorganisms break down organic compounds without oxygen. This document outlines the key topics in fermentation technology including: fermentation processes, microbial rates, stoichiometry of microbial growth and product formation, and important fermentation products. It also provides examples of stoichiometric calculations for microbial growth including determining yields, degrees of reduction, and setting up general stoichiometric equations using conservation principles.
The document describes the development of a high throughput platform for screening glycoside hydrolase enzymes based on oxime-NIMS. Key points:
1) JBEI developed an approach integrating oxime chemistries, acoustic printing and nanostructure-initiator mass spectrometry (Oxime-NIMS) to quantitatively analyze glycan products of GH reactions, including with complex biomass substrates.
2) They defined a standard panel of 12 substrates spanning plant cell wall linkages.
3) The platform was automated and tested by studying cellulases and their synergistic combinations across reaction conditions - generating datasets to enable enzyme cocktail design.
The document describes the principles and design of activated sludge wastewater treatment. Activated sludge processes use aerobic bacteria to consume organic matter in wastewater. Wastewater flows into an aeration tank where bacteria grow as they consume organic matter, then flows into a clarifier where bacteria settle as sludge. Sludge is partially recycled to the aeration tank to maintain bacteria populations. The design of activated sludge systems depends on parameters like hydraulic retention time, solids retention time, food to microorganism ratio, and dissolved oxygen levels. Typical designs include conventional, complete mix, extended aeration, and high rate aeration configurations.
JBEI Research Highlights December 2016 Irina Silva
The document describes research that engineered the 1-deoxy-D-xylulose 5-phosphate (DXP) pathway in the yeast Saccharomyces cerevisiae. Key findings include:
- An engineered strain was able to grow in the absence of mevalonate, reaching 80% of growth compared to strains with mevalonate added.
- The DXP pathway became functional enough to support moderate growth without the mevalonate pathway, but growth on agar without mevalonate was not achieved.
- This work established a foundation for further optimizing yeast to produce bioenergy-relevant isoprenoids through the DXP pathway under industrial conditions.
JBEI Research Highlights - October 2021SaraHarmon4
This document summarizes a study that evaluated the potential for nutrient recovery from wet organic waste processing facilities in California to offset synthetic fertilizer demand. The study found that recovering nitrogen and phosphorus from organic waste streams through anaerobic digestion and separation techniques could meet 11% of the state's nitrogen and 29% of phosphorus fertilizer needs. Recovered nutrients would be in the form of liquid fertilizer, struvite, and compost. The approach provides a foundation for analyzing national-level nutrient flows and recovery potentials from bioenergy production.
Synthetic Biology for Plant ScientistsSachin Rawat
Tools of synthetic biology can be utilised to engineer metabolic pathways to optimize production of secondary metabolites and ligno-cellulose. The presentation describes an approach to develop an artificial positive feedback loop to increase accumulation of cell wall polysaccharides. These will decrease the cost of production of plant-based biofuels, paper and other plant products.
Cell Line Development: Reducing timelines and increasing titres fujifilmdiosynth
Cell line development: Reducing timelines and increasing titres by identification of host cell lines with improved characteristics. To develop a mammalian expression platform which rapidly leads to efficient, robust and high quality biomanufacturing processes
Scalability of Cell Culture Processes in Single-use Bioreactors using Differe...KBI Biopharma
Niket Bubna, Cameron T. Phillips, Sigma S. Mostafa and AbhinavA. Shukla. KBI Biopharma, Durham, NC
253rd ACS National Meeting & Exposition
April 2-6, 2017 • San Francisco, CA
#acsSanFran • www.acs.org/SanFran2017
- The study investigated using arbuscular mycorrhizal fungi (AMF) colonization and tree-herb interactions to enhance growth of legume trees for biofuel production on lead contaminated soil.
- AMF inoculation of legume trees increased overall biomass yield compared to non-inoculated plants under lead stress, likely by retaining lead in underground parts and improving nutrient acquisition.
- Combining AMF with planting legume trees with legume or grass herbs further optimized the system for phytoremediation of contaminated marginal lands and production of biofuel feedstocks.
This document summarizes a microcosm study of a butane-utilizing mixed culture for cometabolic degradation of 1,1-dichloroethene (1,1-DCE). Indigenous and bioaugmented microcosms were constructed using aquifer materials and groundwater from a contaminated site. Microcosms were fed butane and/or 1,1-DCE and degradation was monitored over time. Terminal restriction fragment length polymorphism (T-RFLP) analysis was used to characterize changes in microbial community structure. A biotransformation model was developed and fit experimental data to simulate cometabolic degradation kinetics and inhibition effects. Results showed that indigenous and bioaugmented cultures could degrade butane
Julkipli et al. 2022. Optimization of cephalosporin C acylase immobilization ...julkipliamsyir
This document describes research to optimize the immobilization of cephalosporin C acylase (CCA) using crosslinked enzyme aggregates (CLEAs) technique with chitosan as a co-aggregate. CCA was produced from a mutant strain of Escherichia coli and partially purified using ammonium sulfate precipitation. CCA aggregates were formed by co-aggregation with chitosan and immobilized using glutaraldehyde crosslinking to produce CLEAs-CCA. Response surface methodology was used to optimize the immobilization process by screening influencing factors, determining significant factor levels, and optimizing conditions. The activity of CLEAs-CCA was used as the response parameter to be maximized under optimal immobil
En el marco de la jornada Microalgas, ¿una fuente de petróleo verde?, organizada con IMDEA y celebrada el 8 de abril en EOI, Escuela de Organización Industrial, René H. Wijffels, profesor de la Universidad de Wageningen en Holanda, presenta su trabajo sobre biodiesel producido por microalgas, la factibilidad de este estudio y la biorafinería de las microalgas. Finalmente concluye con la presentación de las diversas fases de investigación hasta llegar a la producción de biocombustibles, alimentos y productos químicos.
Investing in Process Development for Increased MSC Production in Stirred Tank...MilliporeSigma
Interested in developing a robust cell therapy manufacturing platform? In this webinar we will share information in the form of case studies that highlight strategies to optimize your cell therapy production process.
Industry trends in regenerative medicine highlight a critical need for closed cell culture systems that support scalable manufacturing of adherent cell therapies. Typical static in vitro culture methods, however, are often too cumbersome and inefficient to support commercial scale production of mesenchymal stem/stromal cells (MSCs). Single-use stirred tank bioreactor systems are a platform that can address this limitation and have been proven effective for microcarrier-based production of adherent cell therapies. Implementation of optimized process control strategies for parameters such as dissolved oxygen (DO) and agitation rate are key to making an efficient transition from planar culture to stirred tank bioreactors. Herein, a stepwise approach to process development for MSC expansion in a small-scale single-use bioreactor is presented. Case studies focus on strategies to optimize DO control and agitation rates for bone marrow derived MSCs in microcarrier culture, highlighting improvements in process efficiency. In the first case study, the impact different gassing methods have on DO control and whether hypoxic growth conditions affect MSC function are examined. The second case study demonstrates the application of Zwietering’s equation for suspension of solids to overcome scaling challenges often associated with microcarrier culture in stirred tanks. Strategies to further improve the seeding process for bioreactor culture will also be reviewed. Identifying optimal seeding and process control strategies for microcarrier-based bioreactor expansion of adherent cells is paramount for the development of robust cell therapy manufacturing platforms.
In this webinar, you will learn about:
· Process development approaches for production scale-up of mesenchymal stem cells (MSCs)
· Implementing single-use, closed systems for manufacturing cell therapies
· Case studies focusing on strategies to optimize DO control and agitation rates for microcarrier-based cultures
Investing in Process Development for Increased MSC Production in Stirred Tank...Merck Life Sciences
This document discusses process development for manufacturing mesenchymal stem cells (MSCs) using microcarrier-based suspension culture in bioreactors. It outlines challenges in scaling up MSC production, including maintaining optimal hydrodynamic conditions and controlling dissolved oxygen. A case study demonstrates that MSCs expanded at 50L scale on microcarriers in a bioreactor retain their multipotent characteristics. The document also proposes strategies for optimizing a closed seed train to decrease manufacturing risks and costs compared to traditional planar seeding methods.
Optimization of ABE Fermentation from Rice Husk Medium using Clostridium acet...pranavdadhich
A chemically defined medium was optimised for the biomass production of Clostridium acetobutylicum in the fermentor using rice husk as the carbon source.
3. $$$
• 120 million tons of
stover
• Cheap
• Not a food source
…. but difficult to
process
Corn Stover, Agriculture and Agri-food Canada, 2008.
Enzymes
Energy
Cellulose is lying around in abundance
3
4. CBP uses a single
reactor for all
production steps.
The CBP State of the
Art relies on
monoculture-based
processing, which isn’t
reliable
Consolidated BioProcessing will be our
vehicle
4
6. Co-cultures are better than monocultures
Co-cultures allow for specialty, robustness, and are inspired by nature.
6
7. Co-cultures are better than monocultures
Cellulose
Degrading
Bacteria
Co-cultures allow for specialty, robustness, and are inspired by nature.
7
8. Co-cultures are better than monocultures
Cellulose
Degrading
Bacteria
E. coli
extraction
Product
Simple sugars
Co-cultures allow for specialty, robustness, and are inspired by nature.
8
9. CBP with communicating Co-Culture
Cytophaga hutchinsonii:
degrades cellulose, aerobic,
sequenced genome
and transformable
9
10. CBP with communicating Co-Culture
E. coli:
well characterized and produce a
variety of products
Our product: biofuel
10
11. 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
0
2
4
6
8
10
12
14
16
18
20
Time
Population
Bacteria 1
Bacteria 2
Herbert H.P. Fang, Heguang Zhu, Tong Zhang, Phototrophic hydrogen production from glucose by pure and co-cultures of Clostridium butyricum and Rhodobacter sphaeroides,
International Journal of Hydrogen Energy, Volume 31, Issue 15, December 2006, Pages 2223-2230, ISSN 0360-3199
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
Time
Population
Bacteria 1
Bacteria 2
Preliminary Modeling Suggests that Co-Culture can
be Improved with Communication
11
15. Modeling Methods: Dynamic Flux
Balance Analysis
Start with the Flux Balance Analysis
Source: BiGG database, Orth et al. 2011
Av=0
Steady
state
v
Max
μ=wTv
A
Stoichiometric
Matrix
15
16. Then consider extracellular dynamics
Flux
Balance
Model
Extracellular
dynamic mass
balances
Uptake/secretion
kinetics
Modeling Methods: Dynamic Flux Balance
Analysis
Modeling Methods: Dynamic Flux
Balance Analysis
Then consider extracellular dynamics
Flux Balance
Model
Max μ=wTv
s.t.: Av=0
lb < v < ub Extracellular Mass Balances
dX
dt
= μX
𝑑𝑆,𝑃
𝑑𝑡
= v
siX
Uptake/Secretion Kinetics
vs = fs(S,P)
S, P
vs
μ, vp
Modeling Methods: Dynamic Flux
Balance Analysis
Then consider extracellular dynamics
Flux Balance
Model
Max μ=wTv
s.t.: Av=0
lb < v < ub Extracellular Mass Balances
dX
dt
= μX
𝑑𝑆,𝑃
𝑑𝑡
= v
siX
Uptake/Secretion Kinetics
vs = fs(S,P)
S, P
vs
μ, vp
16
17. Then consider extracellular dynamics
Flux
Balance
Model
Extracellular
dynamic mass
balances
Uptake/secretion
kinetics
Modeling Methods: Dynamic Flux Balance
Analysis
Modeling Methods: Dynamic Flux
Balance Analysis
Then consider extracellular dynamics
Flux Balance
Model
Max μ=wTv
s.t.: Av=0
lb < v < ub Extracellular Mass Balances
dX
dt
= μX
𝑑𝑆,𝑃
𝑑𝑡
= v
siX
Uptake/Secretion Kinetics
vs = fs(S,P)
S, P
vs
μ, vp
We Overcame Many Significant Challenges in
Implementing Our Models
Gomez, J.A., Höffner, K. and Barton, P. I.: DFBAlab:
A fast and reliable MATLAB code for Dynamic Flux
Balance Analysis. BMC Bioinformatics 15(1)
(2014).
Valid metabolic
networks
Kinetic
parameters Computationally
feasible
Non-native
pathways
17
19. Naïve Co-Culture to tell us what to improve
E. coli
Salts
Filter paper C. hutchinsonii
humboldtmfg.com
drpinna.com
lbc.msu.eduStandardsingenomics.org
19
27. Pros of Continuous Culture
• Higher productivity
• No turnover time
Cons of Continuous Culture
• Susceptible to genetic instability
• Susceptible to contamination
• Impractical in industry
Changing Focus from Continuous to Batch
Culture
27
28. We used a toxin and antitoxin system to modulate C.
hutchinsonii population size
28
30. C. hutchinsonii E. Coli
AHL
Simple sugars
Circuit design tethers C. Hutchinsonii growth to E. Coli
30
31. Modeling predicts circuit failure and success
modes
Not enough
starting AHL
Threshold level of
E. coli too high
AHL
Simple
sugars
Success: C. Hutchinsonii to E.
Coli ratio improved
Success: C. Hutchinsonii to E.
Coli ratio increased
31
34. Origin of Replication Part Design
• PCR amplified chromosomal
origin of replication (oriC)
• BioBrick and MoClo compatible
• Part# Bba_K1705010
C. hutchinsonii Genome
Cloning Site
oriC
pSB1C3
oriC
34
35. Testing Selection Markers
• Inserted oriC in both CmR and
KanR backbones
• Cm resistance to 10 ug/mL
• Kan resistance at 30 ug/mL
Wild-Type Transformed
35
36. C. hutchinsonii Accomplishments
• Opened new, interesting chassis
• Culture and transformation methods
• Modularized origin of replication (Bba_K1705010)
• Selection with Kan and Chlor
• Future directions
• Native constitutive promoter (Bba_K1705012)
• Native RBS (Bba_K1705011)
36
39. -10
0
10
20
30
40
50
60
0 5 10 15 20 25 30
Value(mAU)
Time(min)
90% Methanol Standards
Methanol Ethyl Palmitate Standard Palmitic Acid Standard 1:1 Ethyl Palmitate:Palmitic Acid
Ethyl
Palmitate
Palmitic Acid
Developed a Method to Detect Biodiesel
39
40. -5
5
15
25
35
45
0 5 10 15 20 25 30 35 40 45
Experimental Samples with 90% Methanol
Not Induced Induced
Fatty Acid Ethyl Esters
(FAEEs)
We developed a method to separate a fatty acid
from its ethyl ester
40
45. • Characterized co-culture
• Designed population control system
• Progress toward implementation
What have we achieved?
45
46. • Characterized co-culture
• Designed population control system
• Progress toward implementation
0
0.2
0.4
0.6
0.8
1
1.2
BBa_J23100 BBa_J23102 BBa_J23107 BBa_J23106
Strength
Anderson Promoter Strengths
Part BioBrick #
C. hutchinsonii Origin of
Replication
BBa_K1705010
C. hutchinsonii RBS BBa_K1705011
C. hutchinsonii promoter BBa_K1705012
alcohol dehydrogenase (adhB) BBa_K1705000
Acyl-CoA diacylglycerol
acyltransferase (aftA)
BBa_K1705001
Leaderless thioesterase (‘tesA) BBa_K1705002
Acyl-CoA synthetase (fadD) BBa_K1705003
pLux BBa_K1705006
luxR BBa_K1705007
luxI BBa_K1705008
luxI LVA BBa_K1705009
RelE BBa_K1705004
RelB BBa_K1705005
What have we achieved?
46
47. • Characterized co-culture
• Designed population control system
• Progress toward implementation
• Collaboration
What have we achieved?
47
48. • Addresses issue of population instability
• Modular system
• New use of cellulosic waste
• Future challenges:
• Scalability
• Genetic instability
Broader Context
48