Oktoberfest Program

Fermentation is Brewing

Manayunk Brewing
12 October 2010
Matthew Snyder

1
 Fermentation Is Brewing
• History of Microbial Fermentation
• Industrial Utilization of Fermentation
• Microorganisms: Yeasts and Cultures
• Brewing process versus Protein Production
process
• Fermentation equipment Brewing vs. GMP
• More on Beer
• Oktoberfest
• Prost!
2
 Fermentation History
• Brewing recognized
as the first form of
fermentation
• „Hymn To Ninkasi‟
2000 BC is first
brewing recipe
• Nomads settled to
farm, cultivated
grain for bread

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 Fermentation History
• Establishment of communities
• Popular in the Roman Empire before wine
• Christian abbeys refined brewing methods
• Beer became a staple of diets through the
Middle Ages
• 15th century saw the establishment of
brewing industry
• Introduction of hops to beer as a
preservative
• Rheinheitsgebot of 1516
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 Fermentation History
• Louis Pasteur
identifies yeast as a
living organism
• Eventually furthered
research into cause of
diseases and germs
• Alexander Fleming
and Penicillium
Notatum
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 Fermentation Applied
• Antibiotic of properties of Penicillin identified
• Penicillin production needs resulted in
development of early “deep fermentation” tanks
• World Wars drove development of Penicillin as
well as other fermented products
• Butyl alcohol and acetone derived from Clostridium
acetobutylicum
• Industrial use of fermentation as a production
method expanded through early 20th century

6
 Fermentation Applied
• Food products and additives continue to be
primary industry for fermentation
• Beverages, milk products, baker‟s yeast,
vinegar, dietary supplements

7
 Fermentation Applied
• Renewable energy sources
• Cellulosic ethanol, algenol, bio-oils

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 Fermentation Applied
• Industrial chemicals
• Organic acids, biopolymers, solvents,
higher alcohols

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 Fermentation Applied
• Wastewater treatment
• Pulp & paper, sewage sludge

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 Fermentation Applied
• Biological catalysts
• Enzymes, alkaloids, steroids, cytokines

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 Fermentation Applied
• Therapeutic medicines
• Recombinant proteins, monoclonal
antibodies

12
 Biotechnology
• 1982 Humulin, first biotech drug
• Microbial fermentation transition to
mammalian cell culture
• Benefits of targeted mAb proteins as
therapeutic agents prompted growth
• mAb production via cell culture exceeds
other methods
• New genetic techniques have led to
resurgence of E.Coli as expression platform
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 Fermentation Basics
• Fermentation – the mass culture of a
microorganism for the creation of a product
• Mostly natural biological activity
• Fermentation Products
• Biomass
• Extraction of metabolic products
• Compound modification – biotransformations
• Production of recombinant products

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 Microorganisms
• Escherichia coli
• Staphylococcus aureus
• Saccharomyces cerevisiae
• Filamentous fungi
• Pichia Pastoris

• Common Properties
• Flocculation
• Attenuation
• Mutation
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 Microorganisms
• Saccharomyces cerevisiae
• Biologically classified as fungi
• Many varieties and strains; S.carlsbergenesis
• Ale – top fermenting
• Active temp. range 10-25°C
• 5-7 day fermentation
• Lager – bottom
fermenting
• Active temp. range 7-15°C
• 1-3 weeks fermentation
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 Microorganisms
• S.cerevisiae properties
• Capacity to produce quality beer
• Ability to ferment wort of varying content –
meaning cheap malt or adjuncts
• High growth rate
• Hybridization
• Expresses all products
through secretion – ethanol,
flavor components, by-
products
• Readily separates from beer
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 Microorganisms
• Yeast propagation
• Larger or more specialized, breweries will
maintain their yeast strains
• Others employ contract laboratories
• Brewers collect and reuse
yeast for up to 12 batches
• New culture from storage
• Proprietary yeasts
• Bottle conditioning poses
problems
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 Microorganisms
• Brettanomyces lambicus
• Wild yeast in brewing a traditional beer style
• Specific to a small region in West Flanders, Belgium

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 Microorganisms
• Escherichia coli
• Genetics and biochemistry well understood
• Much literature on molecular cloning techniques
and vector systems
• Genetic manipulation
• Rapid doubling time
• Grows in any media
• Relatively stable cell
after encoding –
minimal mutation risk
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 Microorganisms
• Escherichia coli
• Expression system of choice in industry
• Various expression strategies can be designed in
• Intracellular / Secretion
• Expression systems can
produce high protein
output
• Induction possible
• Basic purification
processes with
secreted product
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 Microorganisms
• E.Coli Propogation
• Industry divided between maintaining Cell Banks
in house or with contract labs
• Individual inoculation
cultures created for
production batches
• Not viable for reuse
• Patent protection for
E.Coli expression
systems

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 Production Processes
• Upstream
• Selection of feed stock and additives
• Preparation of media or wort, sterilization
• Inoculation or Pitching
• Fermentation
• Aerobic vs. Anerobic
• Controls
• Downstream
• Purification or clarification
• Fill / finish
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 Protein Process
• Seed Scale-Up
• Fermentation and Harvest

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 Protein Process
• Purification
• Fill/Finish

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 Brewing Process
• Standard
industrial
process flow
• Raw material
conversion to
final product
• Defined unit
operations

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 Brewing Process
• Malt preparation
• Steeping, germination, kilning
• Mashing – extraction of malt sugars
• Brewing – boiling of wort
• Cooling/Aeration – preparing the wort to
receive yeast
• Pitching – addition of yeast
• Fermentation – sugar conversion to alcohol
• Racking – beer conditioning and aging
• Finishing – bottling and kegging
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 Brewing Process

• Nutrient feedstock from

various grain sources:
Primarily barley and wheat
malt
• Other additives determined
by desired features of final
beer: hops, fruits, spices,
exotics
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 Brewing Process
• Hops are bittering & aromatic components

• Hops are bittering & aromatic

components
• Added during the boil
• Sterilization through boiling of wort
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 Brewing Process
• Cooling of wort is critical
• Aeration to introduce
dissolved oxygen
• Transfer to fermenter

• Pitching of yeast
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 Brewing Process
• Fill/Finish operations
• Bottling
• Kegging

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 Brewing Process
• Fermentation Stages
• Lag (adaptation) phase
• Growth (respiration) phase
• Kräusen phases
• Sedimentation & Aging phases
• The stages may overlap and the timing and
duration of each stage depends on:
• Type and age of yeast
• Conditions of the wort
• By-products produced
32
 Protein Process
• Fermentation
Stages
• Lag phase
(acclimation)
• Log phase
(exponential)
• Stationary
phase
• Death phase

• Clearly defined 33
 Fermentation Summary
• Grain treatment is closely correlated with
media preparation
• Process parameters are important in both
processes
• Sterility is mandatory for all fermentation
processes
• Protein production process is a subset of the
complete fermentation cycle

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 Fermentation Equipment
• Primarily a tankage based process
• Stainless Steel vessels/systems
• Brewing process uses copper and wood vesssels
• The movement toward single use systems in
the GMP environment is not likely for the
brewing industry
• GMP considerations for Protein processes
and products are missing from brewing
process

35
 Fermentation Equipment
• E.Coli fermenter
• Complex systems
• Monitoring of pH,
temperature, dissolved
oxygen
• Feedback for process
control
• Advanced processes may
require online exhaust
gas testing
• Offline testing for cell
density
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 Fermentation Equipment
• E.Coli fermenter
• Small scale

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 Fermentation Equipment
• E.Coli fermenter
• Agitation system
• Air or gas sparge system
• Cooling/heating
maintenance
• Addition ports and
systems
• Sterile boundary; filters,
min distances
• SIP and CIP
considerations
38
 Brewing Equipment
• Brewing fermenter, homebrewing
• No frills
• Sterile boundary, airlock

39
 Brewing Equipment
• Fermentation Tank Systems
• Cylindroconical
• Traditional Ale Top-Skimming System
• Traditional Ale Dropping System
• Yorkshire Square System
• Burton Union System
• Open Square Fermenters
• Dual Purpose Vessel System
• Oak Cask

40
 Brewing Equipment
• Brewing
fermenter
• Simple systems
• Minimal controls
• Off-line
monitoring
• Designed for
harvest
• CIP at large scale

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 Brewing Equipment
• Brewing fermenter

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 Process Equipment
• Heat Transfer

43
 Process Equipment
• Unique Heat Transfer

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 Process Equipment
• Vessels

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 Process & Equipment
• Clarification /
Filtration

46
 Other Observations
• Fill/Finish and product delivery
• Vial form, injectables, transdermal, inhaled, solid
dose

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 Other Observations
• Fill/Finish and product delivery
• Bottle design, serving glassware

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 Other Observations
• Fill/Finish and product delivery
• Extreme technology

49
 Other Observations
• Contract manufacturing
• Labeling
• Use & Warning

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 Other Observations
• Regulations, Procedures and R&D
• Minimal regulations for composition/efficacy
• For many craft brewers experimenting with new
beers styles or enhancing old styles is their R&D
• Small batches allow for consumer trials and testing

51
 Other Observations
• Regulations, Procedures and R&D
• For many larger or traditional brewers adherence
to time tested methods are key
• Use of same equipment, same process, in the
same location is actually critical for some beers

52
 Beer Styles
• Ales
• Pale, Blonde, Brown, Strong, Porter, Stout
• English Ales
• Mild, Bitter, ESB, IPA, Barleywine
• Belgian Ales
• Dubbel, Tripel, Quadrupel
• Lambic, Geuze, Flanders sour
• Biere de garde, Saison, Wit
• German Ales
• Altbier, Hefeweizen, Dunkelweizen
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 Beer Styles
• American Lagers
• Light lagers, Adjunct lager, Malt liquor
• German Lagers
• Bock, Dobbelbock, Dunkel, Schwartzbier
• Märzen or Oktoberfest
• Pilseners
• Mixed Style
• American Steam Beer / Cream Ale
• Kölsch
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 Oktoberfest
• Royal wedding on12 October 1810
• Tradition continues in 1811
• Beer, as feature, introduced in 1818
• 200th anniversary
• 177th official
celebrations

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 Oktoberfest Facts
• 16 days; 6.3 million people; 98,000 seats
• World‟s largest annual festival

56
 Oktoberfest Facts
• 6.2 million litres of beer served
• 4.5% ABV

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• Prost!

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