This document provides an overview of microbial fermentation processes used in the beverage industry, specifically for wineries and distilleries. It discusses the types of fermentation (e.g. ethanol, lactic acid), microbes involved (e.g. Saccharomyces cerevisiae, lactic acid bacteria), and how fermentation is used to produce different alcoholic beverages like wine, beer, rum and whisky. It also summarizes the key steps in the winemaking process, including fermentation, aging, and the factors that influence fermentation. Current applications and challenges of fermentation technology in Nepal are also reviewed.

1. APPLICATION OF FERMENTATION TECHNOLOGY:
MICROBIAL FERMENTATION OF BEVERAGE-WINERY,
DISTILLERY
Presentation by:
Preyasha Shrestha 34)
Nilima Shrestha (33)
Rabi KC (20)
Himanshu Bhandary (4)
Presentation to:
Ashna Dhakal
(Associate Professor)
Department of Biotechnology
KATHMANDU UNIVERSITY

2. ● Fermentation is a metabolic process that converts sugar to acids, gases
and alcohol.
● It is of different types (Lactic Acid Fermentation, Ethanol Fermentation &
Acetic Acid Fermentation) resulting from the action of yeast and other
microbes.
● The dominant types of yeast used to make beer are Saccharomyces
cerevisiae, known as ale yeast, and Saccharomyces ovarium, known as
lager yeast used in the fermentation.
FERMENTATION

4. MICROBIAL FERMENTATION OF BEVERAGES
● Microbes can be grown in fermenters to make beverages like rum, beer, whisky etc., at
industrial scale.
● Low alcoholic content beverages can be prepared by the fermentation of starch
products, while high alcoholic content beverages can be produced by the distillation of
fermented malted barley, molasses etc.
● Depending on the substrate used for fermentation and the type of processing, different
alcoholic drinks can be prepared.
● Saccharomyces cerevisiae is commonly known as 'brewers yeast' as it produces
alcohol by fermentation of different malted cereals and fruit juices.

5. WINE

6. WINE
➢ Wine is an alcoholic beverage typically made
from fermented grapes juice.
➢ The species that are most widely used in wine
production are Vitis labrusca and especially Vitis
vinifera which has long been the most widely
used wine grape throughout the world.
➢ Wine’s average range of alcohol is about 10%
ABV (alcohol by volume) to 15% ABV. Of
course, there are a few exceptions: Moscato
d’Asti is as low as 5.5% ABV, and Port is
fortified with neutral brandy upping it to 20%
ABV.
➢ Process of fermentation of grapes into
alcoholic beverages. Besides grapes, bananas,
cucumber and other fruits are also used.
➢ During fermentation ,yeast consumes the
sugar present in the juice and converts it to
ethanol, carbon dioxide and heat.
➢ Involves three categories of operations;
Pre - fermentation, fermentation and post
fermentation. In case of grapes, pre-
fermentation involves crushing the fruits and
releasing juice. Then, Yeast is added to initiate
the process of fermentation.
WINE FERMENTATION

7. HISTORY
● The technique of wine making has been known since the dawn of civilization and
has followed human and agricultural processes.
● The earliest known bio molecular archaeological evidence if plant additives in
fermented beverages dates back from the early Neolithic Period in China and the
Middle-East.
● The connection between yeast and the process of fermentation was noted by
Pasteur
● Embeden-Mayorhof-Parnas (EMP) Pathway has contributed to the understanding
of complex chemical process involved in the conversion of sugar into ethanol.

8. Process of Fermentation
HARVEST
BOTTLING & AGING
CRUSHING
FERMENTATION
CLARIFICATION

9. ● Time of harvest impacts the composition of grapes influencing relative levels
of sugar, amino acid content, relative number and species of of microbes.
● Harvested fruits are destemed and crushed where the juice is released.
● Fermentation can be done either by commercially prepared yeasts or yeasts
naturally present in the must.
● Fermentation is considered to be complete if the sugar level drops to a low or
undetectable level.
● During the aging process, microbial-derived compounds such as
polysaccharides, free amino acids, and peptides contribute to the composition
of wine.

10. • White wine is a wine whose colour can be
straw-yellow, yellow-green, or yellow-gold
coloured.
• Produced by the alcoholic fermentation of the
noncolored pulp of grapes which may have a
white or black skin.
• Some varieties are well-known such as the
Chardonnay , Sauvignon, and Riesling

11. How White Wine is made?

12. • The juice from most black grapes is
greenish-white; the red colour comes
from anthocyan pigments present in
the skin of the grape; exceptions are
the relatively uncommon teinturier
varieties, which produce a red
coloured juice.

13. How Red Wine is made?

14. Fermentation type based on types of yeast
Autochthonous Fermentation
● It is conducted by the organism originally present in
the juice or must.
Inoculated Fermentation
● It is conducted by using commercial active dry yeast
or preservatives like sulphur dioxide.

15. Other types of Fermentation
Bottle
fermentation
Carbonic
maceration
Malolactic
Fermentation

16. ● It is a method of sparkling wine production, also known as champagne
method
● The wine is made bubbly due to the containment of carbon dioxide (by-
product).
Bottle fermentation

17. Carbonic maceration
● In this process, whole bunches of uncrushed
grapes are used.
● Fermentation occurs intracellularly.

18. Malolactic Fermentation
● It occurs after the completion of alcohol fermentation.
● It occurs when the concentration of malic acid is high in the fermented wine.
● It occurs due to the metabolic activity of lactic acid bacteria(LAB).
Malolactic enzymes
Lactic acid + carbon dioxide
Malic Acid

19. Factors influencing
Wine fermentation
Temperature
Sugar Content
Acid
Microorganisms
pH

21. Yeast:
It is a microscopic fungus consisting of single oval cells that reproduce by
budding.
Yeast contribution in Wine Fermentation

22. ● Occurrence: Widely distributed in nature and are associated with
Ascomycetes.
● Isolation: Soil of wine yards, fruits and distillery environment.
● Characteristics:
1. High alcohol tolerance ( above 12%).
2. High ethanol productivity and are rich in protein and vitamin-B.
3. High temperature tolerance ( above 40 degree celcius).
4. Broad substrate utilization range.
● As a result of metabolism of yeast, high levels of alcohol are synthesized
during fermentation from oxoacids originating form amino acids and sugar
metabolism.
● Active dry yeasts of Saccharomyces cerevisiae are commonly used since
20th century for the predictable quality of wine.

24. Lactic Acid Bacteria in Wine Fermentation
● The quality of wine is determined by different factors including climate, fruit
variety, production process as well as the use of different strains of yeast and
bacteria.
Metabolic activity of
LAB
Metabolism of citric and amino
acid
Hydrolysis of grape glycosides
Synthesis and hydrolysis of
ester

25. ● In wine, LAB performs a second fermentation consisting of decarboxylating L-
malic acid to L-lactic acid designated by malolactic fermentation.
● MLF helps for microbiological stabilization as well as lowers the wine acidity
and its nutrient content.
● Also LAB have the ability to produce the antimicrobial compounds.
Metabolism of
LAB
Final product
Organic acids
propionic, acetic and
lactic acids

26. DISTILLATION PROCESS
The distillation process is one
of the most important steps
when making certain kinds of
alcohol.

27. WHAT IS DISTILLATION ?
Distillation is a process of separating component substances from liquid mixtures through vaporization
and condensation, based on different volatility of components in the mixture.
● How does distillation work?
The principle of distillation is simple which based upon the
different boiling points of different liquids for separation.
Lets see the example of alcohol distillation;
Alcohol boils at : (78.5 C, or 173.3 F)
Water boils at : (100 C, or 212 F)
If a liquid containing ethyl alcohol is heated to a temperature
above 78.5 C but below 100 C and the vapor coming off the
liquid is condensed, the condensate will have a higher alcohol
concentration, or strength.

28. DISTILLATION
POT STILL PATENT STILL

29. POT STILL DISTILLATION
● A pot still is a type of still
used in distilling spirits such
as whisky or brandy.
● Heat is applied directly to the
pot containing the wash (for
whisky) or wine (for brandy).
● This is called a batch
distillation.

30. PATENT STILL DISTILLATION
How a patent still works
● The first column (called the analyser)
in a column still has steam rising and
wash descending through several
levels.
● The second column (called the
rectifier) carries the alcohol from the
wash, where it circulates until it can
condense at the required strength.
A column still, also called a continuous still, or Coffey still, is a variety of still consisting of
two columns.

31. Challenges of Microbial
Fermentation
Microbial Incompatibility
Management of Malolactic
Fermentation (MLF)
Nutritional Requirements
Microbial Interaction

32. CURRENT STATUS OF WINE
INDUSTRY IN NEPAL

33. HOW THE BEVERAGES PRODUCTION FLOURISHED IN NEPAL?
The alcoholic fermentation of grains to produce alcoholic beverages has been
practised since ancient civilization and people were very much familiar with ‘drink’.
Liquors have religious significance and if trace back to Vedic civilization, there
were two kinds of drink called Soma and Sura.
Soma: Drink offered to
gods and goddess.
Sura: A kind of secular
drink.

34. • At present, the people of
existing Nepali society
consumes three types of
liquors prepared by different
techniques.
• One is traditional liquors
which are brewed at home
(‘Jaand and Rakshi’) which
also have religious
importance.
नेपाली organic beer preparation

35. Second type of drink is manufactured at local distilleries by modern technology .
&
Third type of drinks include the imported brand of beverages which have no socio-cultural and
religious value.
Pataleban: The story of Nepal’s pioneering vineyard Liquor sales surge as Dashain festivities begin

36. ❑ Different communities have different techniques of distillation, fermentation and
filtration for the preparation of local wine and beer.
❑ Even though the grains are same, the quality of wine and beer varies from one
region to another.
❑ Yeast known as “Marcha” in Nepali is mostly commonly used for the process of
fermentation since long ago.

37. https://youtu.be/E3C89iUkhb4 - FERMENTATION distillery animation
https://youtu.be/5ByxeIoQaXQ - DISTILLATION distillery animation

38. CONCLUSION
❖ Fermentation is the future of the alternative
protein industry.
❖ Fermentation could have a role to play in very
large volumes of inexpensive raw materials as it
moves to commercial scale.
❖ There are a lot of opportunities in fermentation
to think about how we biotech engineers use
microbial fermentation to produce large amounts
for the industry.
Fermentation for cultured meat
by Katy Askew, 30 April 2020

39. REFERENCES
1. Pumphrey, B. (1996). AN INTRODUCTION TO FERMENTATION. UK: New Brunswick Scientific Ltd.
2. Liu, D., Zhang, P., Chen, D., & Howell, K. (2019, November 20). From the Vineyard to the Winery. Retrieved
from Frontiers in Microbiology: https://www.frontiersin.org/articles/10.3389/fmicb.2019.02679/full
3. Kaur, P., Ghoshal, G., & Banerjee, C. U. (2019). Fermented Beverage. Science and Technology of Fruit Wine
Production.
4. Saranraj, P., Sivasakthivelan, P., & Naveen, M. (2017). Fermentation of fruit wine and its quality analysis: A
review. Australian Journal of Science and Technology, 97.
5. Nardi, T. (2020). Microbial Resources as a Tool for Enhancing Sustainability in Winemaking.
Microorganisms, 507.
6. Inês, A., & Falco, V. (2018). Lactic Acid Bacteria Contribution to Wine Quality and Safety. Generation of
Aromas and Flavours.
7. Heaton, T., & Prasain, K. (2019, July 13). Nepali winemakers, buoyed by demand, are experimenting and
innovating in local vineyards. Retrieved from THE KATHMANDU POST:
https://kathmandupost.com/food/2019/07/13/nepali-winemakers-buoyed-by-demand-are-experimenting-and-
innovating-in-local-vineyards