Demystifying

Diastaticus
An Exposé of Everyone’s Favorite Explosive Yeast
Matt Linske
Manager & Lead Microbiologist
Brewing and Distilling Analytical Services, LLC
BDAS Denver
What is
Saccharomyces
cerevisiae var.
diastaticus?
• Saccharomyces = Sugar fungus
• cerevisiae = from beer
• diastaticus = separation (of starch)
 When Yeast Super Attenuation
Goes Variant of Beer

• Andrews & Gilliland

• Describe a “novel
species” in 1952
• Later genomic
Variant: taxonomic research (~1985)
designation below reclassified as
species and subspecies S. cerevisiae variant
 What makes diastaticus different?

Starch degradation It’s in their Genes Molecular Machines

Maltotriose
STA1
Maltotetraose
STA2
Extracellular
Dextrin
STA3
Glucoamlyase
Starch
 How it works:
The Central • Signal within cell
Dogma: activates DNA
(gene)
Information • DNA transcribed
transfer within into mRNA
cells • RNA converted to
protein by
Ribosome
STA Gene family is highly
homologous
The Glucoamylase Multigene Family in
Saccharomyces cerevisiae var. diastaticus:
An overview. Pretorius et al. 2008.

DEX allelic to STA genes

Allelism within the DEX and STA gene
families in Saccharomyces diastaticus.
Erratt JA, Nasim A. 1986
Rapid Methods for
Detecting
Saccharomyces
diastaticus, a Beer
Spoilage Yeast, Using
the Polymerase Chain
Reaction
H. Yamauchi, H. Yamamoto, Y. Shibano, N.
Amaya, and T. Saeki, JASBC 1998
Structural analysis of
glucoamylase encoded
by the STA1 gene of
Saccharomyces
cerevisiae (var.
diastaticus)
Adams et al, Yeast 2004; 21: 379–388.
Enzymatic
Action!
 Brewing with diastaticus
Apparent Gravity vs Time Strains:
1.050
1.045 • Brewing Science Institute
1.040
A-18 London Ale III (STA1-)
1.035
1.030 • White Labs WLP566
1.025
1.020
Belgian Saison II (STA1+)
1.015
1.010
• Voss Kveik (STA1-)
1.005 STA1 positive yeast took longest to
1.000
0 1 2 3 4 5 6 7 8
reach terminal gravity, lower FG
than STA1 negative yeast!
BS1 A18 WLP566 Voss Kveik
 Not all
diastaticus
Fermentations
are the same

Saccharomyces cerevisiae variety

diastaticus friend or foe?—spoilage
potential and brewing ability of different
Saccharomyces cerevisiae variety
diastaticus yeast isolates by genetic,
phenotypic and physiological
characterization.
Tim Meier-Dörnberg et al, 2018
 Differences in Fermentation
Related to Different Gene Expression

A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains. Krogerus et al 2019
 Differences in Fermentation
Related to Different Gene Expression

A deletion in the STA1 promoter determines maltotriose and starch utilization in STA1+ Saccharomyces cerevisiae strains. Krogerus et al 2019
Diastaticus as a Contaminant

Locations, Effects, & Detection

What Potential Impacts
happens Over-attenuation
• Final gravity 1.004 or less

when • Flavor imbalance

• Package failure

diastaticus
• Bottle bombs or Can shrapnel =
SAFTEY ISSUE!!!

is
Phenolic Off Flavor
• Not all strains POF+

present? Sediment/haze
Where does contamination occur?
• Brewhouse
• Pipework
• Pitching yeast
• Fermentation cellar
• Packaging lines

Incidence of Saccharomyces cerevisiae

var. diastaticus in the Beverage Industry:
Cases of Contamination, 2008–2017
Tim Meier-Dörnberg et al 2017
What is the level of risk?

Low Cell numbers can still

cause problems!
Best practice: Zero Tolerance
The Viability of Minimal Numbers of Saccharomyces diastaticus
in Beer, Robert P. Greenspan (1966)
 Methods of Detection
Total Polymerase
Plate Chain
Counts Reaction
 % RECOVERY NBB VS. YPD + CuSO4
250 PPM 300 PPM 350 PPM
140.0%

120.0%

100.0%

80.0%

60.0%

40.0%

20.0%

0.0%
 168.49%
59.42%

155.47%
141.82%

140.85%
116.67%

128.43%
0.00%

115.49%
81.73%

114.57%
117.14%

104.41%
100.00%

102.39%
109.62%

102.38%
131.43%

100.00%
151.11%

100.00%
0.00%

98.13%
103.13%

97.24%
13.89%

96.88%
26.53%

96.15%
52.17%
% Recovery 2019 LCSM vs. NBB

95.08%
30.83%

93.53%
83.33%

86.49%
116.00%

59.39%
50.00%

53.01%
78.05%

42.14%
103.70%

38.59%
0.00%

32.48%
5.71%
LCSM: 2018 VS 2019

29.90%
103.03%

11.38%
58.70%
% Recovery 2018 LCSM vs. NBB

8.45%
62.95%

5.67%
0.00%

4.46%
48.00%

0.59%
82.00%

0.00%
0.00%

0.00%
0.00%

0.00%
0.00%
 IOYB64, % RECOVERY VS NBB
FPDM-U FPDM-W LCSM-B LCSM-W
160.00%

140.00%

120.00%

100.00%

80.00%

60.00%

40.00%

20.00%

0.00%
0 24 48 72 96
 Optimizing LCSM For Diastaticus

WLP 565, % Recovery on LCSM STA1+ #20D-0206, % Recovery on

120.00% LCSM
100.00% 120.00%
% Recovery

80.00% 100.00%

% Recovery
80.00%
60.00%
60.00%
40.00%
40.00%
20.00% 20.00%
0.00% 0.00%
0 100 200 300 400 500 600 0 100 200 300 400 500 600
CuSO4, ppm CuSO4, ppm

Yeast #4: 0.5g (NH4)2-SO4 Yeast #4: 0.75g (NH4)2-SO4 Yeast #206: 0.5g (NH4)2-SO4 Yeast #206: 0.75g (NH4)2-SO4
• Enzoklop - CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=32003643
A Primer
on
Primers • Short, single-stranded nucleic acid utilized in
the initiation of DNA synthesis
• Both Forward and Reverse Needed to Amplify
Gene of interest

• By Zephyris - CC BY-SA 3.0,

https://commons.wikimedia.org/w/index.php?curid=26794032
Real
Time
PCR • DNA amplification detected by
fluorescent dyes
• Can be visualized in Real Time!
End
Point
Detection
Thank you!
• Team BDAS
• Chuck Skypeck and BA Staff
• Justin Levaugn
• Jess Caudill
• David Bryant
• Lance Shaner & Laura Burns
• Karen Fortmann & Kara
Taylor
• Spencer Weeks
• Tom Boudreau & Dr.
Matthew J. Farber