Practical Propagation

MBG/MBAA
2011 Winter Conference
Kalamazoo MI

Why you dont you want to implement

any of the information in this talk?

Expensive!
Time intensive.
Trial and error optimization required
Stringent sanitation and QC is a requirement.

Yeast Source Options

From Prior Brews

Receive from a friendly local brewery
Dried Yeast
Purchase Pitchable & Propagate (in tank)
Grow from slant in house, propagate in tank
Complete Single Cell to Plant Scale
Propagation Program using Dedicated
Propagation Equipment.

Slurry vol
1600 ml
Slurry conc.
2,500 mcells/ml
Total cells 4,000,000 mcells
Desired Pitch conc
8 mcells/ml
Calculated wort vol.
5.00 hl
Wort vol
Pitch conc

7 bbls
821 liters
4.87 mcells/ml

Dilute 4X at:
3o P drop,
2 days or
30-40 X 106 c/ml

Drauflassen
flowing on to
A portion of the propagated media is left behind
and fresh wort is added. The propagation starts
again and desired cellular densities are achieved
rapidly.
Blend off Draufloss at 10% or less

Why didnt it work for us?

Insufficient quantities from appropriate sources

6 Yeast source beers

2 Stouts (too dark)
4 Types have excessive starting Plato
1 Style Micro-compromised (Dry Hop)
5 Products use other yeast strains
Developed Logical Test Spreadsheet for Sales Dept.

Overall: Not fast enough, Not enough yeast,

Potential flavor matching issues

Louis Pasteur

Gooseneck Flask

Propagation Goals
Acceptably high cell counts
High viability (>95%)
Good vitality or metabolic activity with (high
glycogen and sterols levels)
Free of contaminating microorganisms
110-150 mcells/ml of healthy yeast
Yeast Health is more important than Cellular
Mass

Yeast Nutritional Needs

Sugar source
FAN source (trub, yeast, wort)
O2
Trace elements: Zn, Cu, Mg,

Propagation Considerations

Propagation Temperature
Sugar Spectrum
Wort: 7-12oP
Yeast Nutrient
Low CO2
Aeration (with air!)
Nucleation sites

Sterile Air/O2 (0.22um filter)

Linseed Oil
Foam: Antifoam, 50% headspace or defoamers
0.5ppm Zn (zinc sulfate heptahydrate)

Flavor Concerns
Oxidized flavors
Esters
While yeast is in growth phase it consumes more
acetyl CoA (ester precursor)

Blending may be required

Dilution Ratios
Why?
6 X rule
Exceptions

Sample Methods: Stepped Propagation

100 ml 1 liter 15 gal 2 Bbl 8 Bbl 50
Bbl
Diluting to 10 mcells/ml at each move
10 ml 1 liter 1 Hl 5 Hl 18 Hl 200Hl
E. flask C. Flask 2.5 Bbl 10 Bbl ?
4X volume increase at each move
Each is a 7 day ferment, so X weeks to prop up
Mid-Plato wort & 12 ppm of O2 at each addition

Identifying the Ideal Yeast

Propagation Technician:

Diagnosis: Paranoid
with Obsessive
Compulsive
Disorder
(manifested through
continuous cleaning
behavior)

Sanitation critical
Bacterial growth rate is much (4X) higher than
yeast
Propagator maintains ideal growth conditions
Ample sugars, high pH, high O2, low CO2 etc.
Labwork to verify that propagation is clean
Differential Media Plating & ATP PCR
verification

Labor Growth
Pitch
Time Time
Task
Container
Volume
Ratio
1
72 Prep & streak
Culture plate
1
36 Innoculate 90 ml Vial
0.5
48 Innoculate 500 ml Flask
6.6 :1
1.5
36 Innoculate 20 liter Carlsberg Flask
40 :1
0.25
24 Innoculate
7 Bbl Prop Tank
42.1 :1
0.25
24 Innoculate 18 Bbl Prop Tank
3.6 :1
2
Micro Plating & Assay

6.5

240 Total (Hours)

Task Time
Labwork to prop to 20l 390
Sanitize/Setup Prop System 120
Wort to Wort Tank 30
Panel/Program Changes 50
Wort & Inoculum to Prop Tank 40
Topping Wort Charge #1 30
Pitch 30
Clean 270
(16 Hrs) Total 960

Purchase 7 Bbl Pitchable Slurry

$
258
+ Shipping

Labor: 16 hours @ $22/hr =

Wort & Lab Supplies

$
$

Propagator

$120,000

352
?

Equipment Design & Operation

Simple, stepped size fermentations

Pumping/mixing
O2 Additions/Control
Substrate (wort) Additions/Control

Carlsberg Flask
25 liter net
sterilize with
heat
pitch with 200
ml lab prop
aeration lance
~ $6,000

Respiration, Fermentation,
Crabtree & Glucose

Glucose > 0.4% induces Crabtree effect

As low as 0.2 g glucose/liter?
Incremental Feeding (Zulauf Process)
Used for Bakers yeast propagation
Up to 2.5 X as much yeast as batched process
12oP is 120g extract in 880g water
10-15% of extract is glucose/fructose

Fed Batch Difficult: must Control pH, O2 & Sugars

Newlands YPS

60, 120, 240 liter systems

$26-38,000
Operates as incremental feed system
As yeast consumes sugars in the wort more is
constantly added
Ideally, the propagation slurry Plato stays low
and the cellular concentration is high.

Starter Volume
Cell concentration
Wort Addition
Total Volume
Pitch rate

120 liters
120 mc/ml
15 Bbls
1880 liters
7.7 mc/ml

Frings

1.25, 25, 200 hl working volumes

Aeration device, Mechanical defoamer
Vessel DO controlled to ~ 2ppm
Growth of up to 12 X in 24 hrs
Cell count of 150-200 X 106 /ml possible
Pitching rate of 1-1.5 l/hl
~$80-90,000 for 25 Bbl (net) system

Esau Huber
Alfa Laval/Scandi Brew
$100-150K for 5-30 hl
swing panel, pump, plc,
Mixers, Pumps, Aeration Lance, Turbojet

Labor: Continuous Propagation

Task
Labwork to prop to 20l
Sanitize/Setup Prop System
Wort 1 to Wort Tank
Panel/Program Changes
Wort & Inoculum to Prop Tank
Wort Charge #1A
Wort 2 to Wort Tank
Pitch #1

Time
390
120
30
50
40
30
30
30

Total: 1230

Wort Charge #2
Wort 3 to Wort Tank
Pitch #2
Wort Charge #3
Wort 4 to Wort Tank
Pitch #3
Wort Charge #4
Pitch #4
Clean

30
30
30
30
30
30
30
30
270

Hours: 20.5

Pitch 350 Bbl/day, 4 cycles

Cell concentration
Wort to pitch

126 mc/ml
350 Bbls
41,067 liters

Desired Pitch rate

Pitch Volume

6.5 mc/ml
2,119 liters
18.06 Bbls

Starter Volume
7 Bbls
Cell concentration 126 mc/ml
Wort Addition 18 Bbls
Pitch rate 35 mc/ml