This document discusses inoculum development and production media for fermentation processes. It defines inoculum as a culture of microbes used for industrial fermentations that is in an active growth phase and free of contamination. It describes the basic components of industrial fermentation media including carbon, nitrogen, salt, and vitamin sources. It also discusses various methods for preserving microbial cultures long-term, including subculturing, desiccation, freeze drying, and cryopreservation. The key aspects of inoculum development are preparing a seed culture in sufficient quantities and a healthy state to maximize productivity and yield in large-scale fermentations. Examples of media used for specific inoculum development are also provided.

1. SRI PARAMAKALYANI COLLEGE
( Reaccredited with A+ Grade with a CGPA of 3.39 in the III Cycle by NAAC
Affiliated to Manonmaniam Sundaranar University, Tirunelveli
ALWARKURICHI 627 412 TAMIL NADU, INDIA
POST GRADUATE & RESEARCH CENTRE - DEPARTMENT OF MICROBIOLOGY
(Government Aided)
IV SEM - CORE – FERMENTATION AND INDUSTRIAL MICROBIOLOGY
UNIT – 2
INOCULUM DEVELOPMENT AND PRODUCTION MEDIA
K.ARUL THIVYA
REG NO : 20211232516104
II M.SC.MICROBIOLOGY
ASSIGNED ON: 01-03-2023
TAKE ON : 20-03-2023
Submitted to,
Dr.S.VISWANATHAN, Ph.D,
ASSISTANT PROFESSOR & HEAD,
SRI PARAMAKALYANI COLLEGE,
ALWARKURICHI.

2. SYNOPSIS
• INTRODUCTION
• DEFINITION OF INOCULUM
• INDUSTRIAL FERMENTATION MEDIA
• PRESERVATION OF CULTURE
• INOCULUM DEVELOPMENT
• EXAMPLES OF INOCULUM MEDIA USED IN
VARIOUS INDUSTRIAL PRODUCTION
• REFERENCES

3. INTRODUCTION
• Production and yield of fermentation product
depends upon number of factors that controls
fermentation process directly or indirectly
• A good fermentation always start with a good
fermenting strain and culture of such strain in
appropriate density and healthy state.
• Therefore successful fermentation process start
with a excellent inoculum development to obtain
maximum possible for productivity and yield.

4. DEFINITION OF INOCULUM
• Inoculum we use for industrial fermentations
• Inoculum is the mixture of cultured microbes
along with inwhich it is growing.
• Transfer about 0.5-5% inoculum.In its active,
healthy, and exponential growth phase.
• Available free of contamination required large
volumes.
• Retain its capability of the formation of
desired product formation.

5. INDUSTRIAL FERMENTATION MEDIA
• The basic ingredients of media carbon source,nitrogen
sources, inorganic salts, growth factors and distilled
water etc.,
• Maintain of pH in the optimum range is necessary for
making the process successful. If Buffer (e.g. CaCO,) is
the solution to maintain the correct level of pH of the
medium.
• Foaming is the serious problem in a fermentation
industry.
• Hence, defoamers (eg. oil mixed with Octadecanol for
penicillin fermentation) should be used for controlling
foam

6. KINDS OF MEDIA

7. MEDIUM DEVELOPMENT
• To maintain economic competitiveness, low
cost crude material are frequently used.
• Levels of minerals and growth factor may be
critical.
• SOURCES:
C + E + N +02
BIOMASS + PRODUCT +
BY-PRODUCTS + CO2 +
WATER + HEAT

8. FERMENTATION MEDIANUTRIENTS
NUTRIENTS RAW MATERIALS
CARBON MOLASSES, STARCH
NITROGEN
CONSTIPLIQUER , SOYBEAN MEAL ,
PURE AMMONIA OR AMMONIUM
SALTS , UREA , NITRATE SALTS ,
PHOSPHATE SALTS.
VITAMINS
BIOTINE , YEAST EXTRACT , BEEF
EXTRACT
GROWTH FACTORS CONSTILIQUER , WHEAT GERM MEAL.

9. CARBON SOURCES
Starch (corn, wheat, potato)
• Widely used in fermentation industry
• Starch dextrin glucose
• Advantages: cheap than glucose

10. CARBON SOURCES
Molasses
• By-product of cane or beet sugar production
• A dark viscous syrup containing 50%-
75%fermentable sugars (mainly sucrose) with
2%nitrogen, vitamins and minerals
• Cheaper

11. CARBON SOURCES

12. NITROGEN SOURCE
Inorganic nitrogen source
• Ammonia. Ammonium and nitrate
• Microbes can utilize it faster
• After it is utilized, the pH of medium will be
changed.

13. NITROGEN SOURCES
Organic nitrogen source
• Urea
• Yeast extract
• Peptones
• Corn steep liquor
• Soybean cake powder
• Peanut powder
• Brane hydrolysis liquid

14. TRACE ELEMENT
• Fe
• Mn
• B
• Zn
• Mo
• Cu

15. GROWTH FACTOR
• Small amount of organic compounds
necessary for microbial growth
• eg, amino acids, vitamins, biotin
• SOURCES: Normally organic nitrogen source.
• eg com steep liquor

16. CULTURE PRESERVATION
• Industrial important microorganism have the
capability to produce high yields of desirable
metabolites in large scale-up production
fermentation.
• Highly productive mutant strains are preserved
for long periods free from phenotypic change.
• With particular respect to the capability of high
production of a primary or secondary metabolic
product

17. CULTURE PRESERVATION
Four major classes of preservation procedure
are
1. Subculturing or active slant transfer
2. Desiccation in soil or on porcelain beads
3. Freeze drying or Lyophilization
4. Cryopreservation

18. SUBCULTURING OR ACTIVE SLANT
TRANSFER
• Once the available substrate surface is covered by
cells , growth slows and then ceases.
• Necessary to subculture them at regular intervals.
• In order to keep the cells healthy and actively
growing .
• Convenient and inexpensive (does not require
special equipment).
• Followed by incubation at a suitable temperature.

19. DESICCATION IN SOIL OR ON
PORCELAIN BEADS
Many spore forming fungi and streptomycetes
can be preserved can be preserved by drying the
spores on the surface of various insert solid
substrates such as soil, silica gel or glass beads.

20. FREEZE DRYING OR LYOPHILIZATION
• Broth culture or cells harvested from slant
culture are dispensed into tubes or vials
stored in frozen in an ordinary freezer with
temperature ranging from 5-20°C.
• The viability of many microorganism can be
maintained for 1 to 2 years

21. CRYOPRESERVATION
• Ultra cold Temperture freezing(-60-80°C)
• For long term preservation.
1.Mechanical freezers(-80°C)
2. Liquid nitrogen ferrers(-156-196°C)

22. INOCULUM DEVELOPMENT
• The development of active logarithmic microbial
culture that is suitable for the final industrial
production level is known as inoculum development.
• To obtain seed culture in appropriate proportion and
healthy state that gives maximum productivity and
yield.
• This is usually done using flask cultures ranging in
between 50 ml to 12 It and the volume of the Bask or
container can be increased as per the need.
• The volume of inoculum we add to fermenter should
be about 5% to that of media volume.

23. INOCULUM DEVELOPMENT FOR
BACTERIAL CULTURE

24. INOCULUM DEVELOPMENT

25. EXAMPLES OF INOCULUM MEDIA
• Composition of bennett’s medium used in the
preparation of inoculum for vitamins
productions
COMPONENTS AMOUNT (g/l)
Yeast extract 1.0
Beef exttract 1.0
N-Z-Amine A 2.0
Glucose 10.0
Distilled water 15.0

26. CRITERIA FOR GOOD INOCULUM
• Healthy, active state-minimise lag period
• Available in sufficient quantities
• Suitable morphological form
• Free from contamination

27. REFERANCE
• Book Title: Industrial Microbiology
• Author: AH PATEL Edition
• First edition 1985
• Online: https://www.imedpub.com scientific-
journals-list » inocu
• https://www.sciencedirect.com/topics/engine
ering/inoculum-development

31. THANKS
TO