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
IV SEM - CORE – FERMENTATION AND INDUSTRIAL MICROBIOLOGY
UNIT – 2
INOCULUM DEVELOPMENT AND PRODUCTION MEDIA
REG NO : 20211232516104
ASSIGNED ON: 01-03-2023
TAKE ON : 20-03-2023
ASSISTANT PROFESSOR & HEAD,
SRI PARAMAKALYANI COLLEGE,
• DEFINITION OF INOCULUM
• INDUSTRIAL FERMENTATION MEDIA
• PRESERVATION OF CULTURE
• INOCULUM DEVELOPMENT
• EXAMPLES OF INOCULUM MEDIA USED IN
VARIOUS INDUSTRIAL PRODUCTION
• 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.
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
• Retain its capability of the formation of
desired product formation.
INDUSTRIAL FERMENTATION MEDIA
• The basic ingredients of media carbon source,nitrogen
sources, inorganic salts, growth factors and distilled
• 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
• Foaming is the serious problem in a fermentation
• Hence, defoamers (eg. oil mixed with Octadecanol for
penicillin fermentation) should be used for controlling
• To maintain economic competitiveness, low
cost crude material are frequently used.
• Levels of minerals and growth factor may be
C + E + N +02
BIOMASS + PRODUCT +
BY-PRODUCTS + CO2 +
WATER + HEAT
• Small amount of organic compounds
necessary for microbial growth
• eg, amino acids, vitamins, biotin
• SOURCES: Normally organic nitrogen source.
• eg com steep liquor
• Industrial important microorganism have the
capability to produce high yields of desirable
metabolites in large scale-up production
• 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
Four major classes of preservation procedure
1. Subculturing or active slant transfer
2. Desiccation in soil or on porcelain beads
3. Freeze drying or Lyophilization
SUBCULTURING OR ACTIVE SLANT
• 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
• Convenient and inexpensive (does not require
• Followed by incubation at a suitable temperature.
DESICCATION IN SOIL OR ON
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.
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
• Ultra cold Temperture freezing(-60-80°C)
• For long term preservation.
2. Liquid nitrogen ferrers(-156-196°C)
• 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
• 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.
EXAMPLES OF INOCULUM MEDIA
• Composition of bennett’s medium used in the
preparation of inoculum for vitamins
COMPONENTS AMOUNT (g/l)
Yeast extract 1.0
Beef exttract 1.0
N-Z-Amine A 2.0
Distilled water 15.0
CRITERIA FOR GOOD INOCULUM
• Healthy, active state-minimise lag period
• Available in sufficient quantities
• Suitable morphological form
• Free from contamination