Team 1 Technical Writing and

Presentation

Saccharomycodes ludwigii - The maltose negative yeast in non-alcohol

beer brewing
Nguyen Quang Khanh, Hoang Phuong Anh, Nguyen Van Chinh

Abstract

Non-alcoholic beer is a secondary product of the beer brewing industry. We are living in an era that
requires a demanding lifestyle of health is being cared for and directed in the brewing industry. In
which the biological method of using nonconventional yeast species is being studied, adapted for
application in this field to reduce the ethanol content during processing.

Non-alcoholic beer

Non-alcoholic beer is beer that does not contain alcohol or contains very little alcohol. Depending on
the law, the amount of alcohol regulated in different countries is different. In Belgian, beer with alcohol
content <0.5% v/v is called non-alcoholic beer, and the range of alcohol content between 0.5% - 1.2%
is called low-alcohol beer. However, in Italy or France, the regulation on non-alcoholic beer is applied
to beer products with alcohol content <1.2% v/v [1].
There are two main groups of methods for creating alcohol-free beer production: physical and
biological. Some of the physical techniques are osmotic distillation, reverse osmosis, vacuum
rectification and evaporation, and spinning cone column distillation. Based on the ability of substances
to evaporate at different temperatures, manufacturers conduct temperature control to separate alcohol
components from ordinary beer. However, in this process, some aromatic and volatile compounds also
disappear. To overcome that disadvantage, biological methods are commonly applied, especially the
use of non-conventional yeasts species in beer fermentation [1].

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Brewing process

Figure 1. Schematic diagram of the brewing process and the involved raw materials [2].
Beer is a fermented alcoholic beverage using 4 ingredients: water, malted cereal, hops and yeast
(Figure 1). Water is the main material, around 94% of a beer’s weight. Malts are results from
germination, of any cereal such as rice, corn, wheat. Hops are the green cone-shaped flowers of the
Humulus lupulus plant. Yeast is a eukaryotic, single-celled organism that belongs to the kingdom fungi
[2].
The brewing process includes 9 steps, which are: milling → mashing → wort filtration → wort boiling
→ wort treatment → fermentation → maturation → filtration → pasteurization [2]. Milling is the
physical crushing of malt kernels into smaller particles that prepare for mashing [3]. Mashing is the
process where crushed grains are mixed with water in a ratio of 1:3 (w/w) to form a mixture called the
“mash” [3, 4]. After mashing, wort filtration is carried out in a lauter tun, washed with water to increase
sugar extraction and the yield of the process [2]. Boiling includes several purposes, such as sterilization
to eliminate bacteria; production of bitterness in beer; extraction of aromatic hops compounds that
contribute to the taste and aroma of beer [2]. Wort treatment is the consequent step to wort boiling,
which allows the removal of precipitated hops and proteins, cooling and aeration [2]. Yeast is added
when the wort is transferred to a fermentation tank and converts the sugar into ethanol, carbon dioxide
plus some amount of glycerol and acetic acid [2]. Clarification arms to remove solids from wort to give
a clear liquid. The main purpose of clarification is to increase the retention of yeast cells,
macrocolloids, suspended matter and solutes that cause the formation of beer haze [2]. Pasteurization is
the process of heat treating beer to inhibit fermenting yeast and potential spoilage microorganisms
which would otherwise promote undesirable chemical reactions, thus prolonging the shelf life of the
beer [2].

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Maltose nagative yeast

Maltose-negative yeast refers to yeasts that cannot ferment maltose due to deficiencies in the maltose
transporter and maltase enzyme. These secial yeasts are important in non alcohol beer brewing because
they retain a part of the volatile elegance of conventional beers but may also be practical in the removal
of a “worty” taste [5].
Saccharomycodes ludwigii is known as a maltose negative yeast in the production of non alcohol beer.
Beer produced with this yeast strain can have a low ethanol level and fruitier, estery flavours.
Morphologically, it appears as lemon shape (Figure 2), and it presents asexual reproduction by bipolar
budding [6].

Figure 2. Apiculated cells of Saccharomycodes ludwigii at a magnification of 600× [6].

S.ludwigii has been isolated in cases of stuck or sluggish fermentations or during storage of wines. It
has also been detected in fruit juices and their fermented derivatives, tequila and mezcal, in soil
samples, insects and tree secretions. Another source of S.ludwigii is palm sap, from which a fermented
drink known as “palm wine” is obtained in Cameroon [6].
Application of Saccharomycodes ludwigii for fermentable
Beer fermentation is the process of converting sugar in the wort into alcohol and releasing CO 2 through
the live activity of yeast. In addition, some aromatic compounds, volatile compounds are also formed
during fermentation. This is an important process that determines the quality of the beer product.
Table 1. Sugar utilization by Saccharomycodes ludwigii during fermentation [1].

Fermentable sugars Saccharomycodes ludwigii

Glucose +

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Galactose -
Sucrose +
Maltose -
Lactose -
Raffinose +
Trehalose -
Melibiose -
+ Positive; - negative

Ethanol obtained by fermentation

Table 2. Isolation source and isolation locality of the test species [1].

Strain Isolation source Isolation locality

DBVPG 3010 Grape must Italy

DBVPG 3398 Tea beer Not available

DBVPG 3931 Grape must Italy

Three strains of S.ludwigii were selected based on their geographical and ecological origin and are
listed in Table 2. All the strains are conserved in the Industrial Yeast Collection, DBVPG (University of
Perugia, Italy) [1].

DBVPG 3931 1.24

Strains

DBVPG 3398 0.72

DBVPG 3010 0.51

0 0.2 0.4 0.6 0.8 1 1.2 1.4

Ethanol (% v/v)

Figure 3. Ethanol obtained by fermentation with S.ludwigii strains (Redraw from G. De Francesco

et.al, 2015).

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According to Figure 3, most of the S.ludwigii strains produced <1% v/v ethanol; among them, strain
DBVPG 3010 showed the lowest ethanol concentration (0.51% v/v) and strain DBVPG 3931 made
similar levels of higher alcohols to a standard beer (1.24 %v/v).

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Conclusion

S.ludwigii was identified to be a worthwhile yeast species for low-alcohol and alcohol-free beer
production. In particular, the beer produced using DBVPG 3010 showed the lowest ethanol content
(0.51% v/v).
References

[1] Giovanni De Francesco, Benedetta Turchetti, Valeria Sileoni, Ombretta Marconi, Giuseppe
Perretti, "Screening of new strains of Saccharomycodes ludwigii and Zygosaccharomyces rouxii
to produce low-alcohol beer", Wiley Online Library, 2015.

[2] Gonçalo Carvalho, Ana Catarina Leite, Rita Leal, Ricardo Pereira, "The Role of Emergent
Processing Technologies in Beer Production", MDPI, 2023.

[3] "The Oxford Companion to Beer definition of Milling", [Online]. Available:

https://beerandbrewing.com/dictionary/VWpGLEPUS6/#:~:text=Milling%20is%20the
%20physical%20crushing,See%20lautering%20and%20mashing. [Accessed 27 7 2023].

[4] "The Oxford Companion to Beer definition of Mashing", [Online]. Available:

https://beerandbrewing.com/dictionary/BkD3Ilopul/#:~:text=Mashing%20is%20the%20term
%20given,soluble%2C%20creating%20the%20sweet%20fermentable. [Accessed 27 7 2023].

[5] Selin Yabaci Karaoglan, Rudolf Jung, Matthew Gauthier, Tomáš Kinčl, Pavel Dostálek,
"Maltose-Negative Yeast in Non-Alcoholic and Low-Alcoholic Beer Production", MDPI, 2022.

[6] R. D. V. Mantilla, "Saccharomycodes ludwigii, control and potential uses in winemaking

processes", MDPI, 2018.