development of microbial bioproducts
TRANSCRIPT
Development of microbial bioproducts:Characterization, formulation and scale up
Freddy Boehmwald , PhD.Bioproducts Development Coordinator
UC Davis Chile
Bioprocess
• Any process that uses complete living cells ortheir components (e.g., bacteria, enzymes) toobtain desired products for commercial use
Bioproduct
• Materials, chemicals and energy derived fromrenewable biological resources
Renewable
Addingvalue to wastes
Compatible with
industralprocesses
Improvedenvironmentalsustainability
Higherefficiency
Why bioproducts?
Long lastingeffects
Pharmaceutical
Agriculture
Industrial
Biomaterials
Bioenergy
Biochemicals
Diversity of bioproducts
MicrobialBioproducts
Microbial Bioproducts for agriculture and industrial applications
Functionalproducts
Biofertilizers
BiostimulantsBiopesticides
Metaboliteproduction
Enzymes Pigments and small molecules
Microorganisms Single Cell Consortia
División de Bioproductos en UC Davis Chile y su articulación con el ecosistema de innovación en Chile (R4D).
A. Proveer una plataforma costo-efectiva para elbiodescubrimiento de nuevos microorganismos con actividadesespecificas o nuevas actividades para el mercado industrial yagronómico.
B. Caracterización de bioproductos y optimización debioprocesos para el mercado industrial y agronómico.
C. Diseño de procesos de escalamiento, formulación yempaquetamiento para nuevos bioproductos y/o bioprocesos.
Screening & Biodiscovery
Lab scaleproduction
Feasibility I
Pilot scale production
Feasibility II
Market
entry
ProductPrototype
Commercial
Product
Life cycle of microbial bioproducts
functionality
productivity
Is the screening and
characterization methodology
valid?
Is the development strategy in an
agreement with my business
model?
1-2 years
2-4 years
Screening & Biodiscovery
Lab scaleproduction
Feasibility I
Prototyping microbial bioproducts using and R4D approach
Market needs
Defyning COP
Genomics &
Metagenomics
Func.
Assays
Lab and/or field testing
Benchmarking:
Testing against commercial
products
PROTOTYPECOP: Critical operation parameters
Prototyping microbial bioproducts using and R4D approach
Market ideal
COP
Prototype
COP
Products in themarket
•Functionality
•Compositions
•Operative ranges
•First approach to shelf life
•First approach to formulation
•Process environmental impact
•Stability
•Documentation of Lab scale production
•Documentation of the product:
Datasheets and MSDS
Prototypedefinition
Regulation and registration could be a very complex issue and should be
adressed properly in the product development time line.
Licensing
prototypes
MARKET NEEDS
Intellectual
Property
Pilot scaleproduction
Feasibility II
Market
entry
Towards a Commercial Bioproduct DevelopmentPrototype Candidates
Marketing, Distributionchannels, logistics, packaging.
Engineer Heuristics: • Magnitude order• Design
Field testingConsumer testing, Stability, shelf life
COMMERCIAL PRODUCT
EXECUTE
Feasibility II
• New facilities?
• Production in new facilities or existent plant? New plant?
• Requiere ammounts of raw materials and utilities?
• Capital investment
• Manufactoring cost?
• Optimum batch size
• Single batch time
• Amount of product generated per year
• Bottle necks: steps or resources?
• Environmental impact
Towards a Commercial Bioproduct Development
Example 1: Process development for the upscalingof an intracellular and extracelullar lignocelulosic
enzymes.
Determining key stages for a economical viable bioprocess
PURIFICA
TION EN
ROJO
ENZIME
NO MAS
Result Description IndicatorAdequate infrastructure Adaptation of the infrastructure, ie the
scaling laboratory and complete installationof the bioreactor.
Enable scaling laboratory
Purchase and bioreactor installation completed
This result is the finished installation of the bioreactor in the scaling laboratory
Bioreactor installed
Training conducted Completed training in the use of bioreactor Training completed
Commissioning and trial run complete Completed commissioning and trial run of thebioreactor.
Fully operational bioreactor
Fermentation 1 (batch) Increase cell mass produced per liter of cultureof the microorganism 1, regarding theperformance obtained at laboratory scale.
Performance greater than 25 g cell / L of culture
Fermentation 2 (feed-batch) Increase cell mass produced per liter of cultureof the microorganism 1, regarding theperformance obtained at pilot plant scale.
Performance greater than 50 g cell / L of culture
Purification intracellular Enzyme from microorganism 1 Superior process efficiency to 500 mg (enzyme) per liter of culture
Purification extracellular Enzyme from microorganism 2 Superior process efficiency to 800 mg (enzyme) per liter of culture
Standard operating procedures (SOP) Development of a document that contains thestandard operating procedures.
Document existing and operationally proven
Quality control procedures (QA/QC) Preparation of a document that containsstandardized procedures for quality control
Document existing and operationally proven
SolventextractionINTRACELLULAR
PRODUCTS
EXTRACELLULAR PRODUCTS
Celular lysis
Sonicator
Beat beater
Frech Press
Enzyme 1 purification
Detergentextraction
Centrifugation
ChromatographicSeparation 1
FPLC
HPLC
Metaboliteconcentration
Ultrafiltration
Precipitation orsolvent extraction
Rotary evaporation
SupernadantMetaboliteconcentration
Ultrafiltration
SolventPrecipitation
(NH4)2SO4 Precipitation
ChromatographicSeparation 2
FPLC
HPLC
Metaboliteconcentration
Ultrafiltration
Precipitation orsolvent extraction
Rotary evaporation
Product Shelf Life
and Stabilization
Lyophilization
Frozen (-80ºC)
LIquid
1.11.2 1.3 1.4 1.5
1.6 o 2.5
2.1 2.2 2.32.4
2
Pellet obtention
1
0
Biomass concentration
800 L Bioreactor
Description of the production procecess Downstream
Bottlenecks
Optimization
Clarification
Synthesis of bioseparation and recovering processes: As the limiting step in bioprocess development
Considerations:
“Experienced engineers heavily rely on heuristics for the design of a new recovery and purification process:
1. Remove the most plentiful impurities first
2. Remove the easies to remove impurities first
3. Make the most difficult and expansive separation last
4. Select processes that make use of the greatest diffetences in the properties of the product and its impurities
5. Select and sequence processes that expliciti different separation driving forces.”1
1.Petrides, D. (2000). Bioprocess Design. Scotch Plains. INTELLIGEN, INC.
May represents up to 20 operation units in the process!
BIOPRODUCTOSResearch for Development model (R4D)
En un mercado que se acerca a la madurez, la diferenciación de losproductos es fundamental para el posicionamiento de la compañía.
En un mercado cada vez mas competitivo, los costos de producciónson fundamentales para un precio competitivo en un mercadoglobalizado.
En un mercado cada vez mas regulado, la sustentabilidad de labioproducción y los controles de calidad en el producto final tienen unalto impacto en la entrada a nuevos mercados.
Muchas Gracias
Freddy Boehmwald , PhD.
Bioproducts Development Coordinator
UC Davis Chile