SHREE RAMKRISHNA INSTITUTE OF

1 COMPUTER
EDUCATION AND APPLIED SCIENCES

NUTRIENT RECYCLE &

BUFFERS
PRESENTATION BY : KALSARIYA TWINKAL
2 INTRODUCTION
Most fermentations require liquid media, often referred to as
broth, although some solid-substrate fermentations are operated.
Fermentation media must satisfy all the nutritional requirements
of the microorganism and fulfil the technical objectives of the
process.
The nutrients should be formulated to promote the synthesis of
the target product, either cell biomass or a specific metabolite.
 Where biomass or primary metabolites are the target product,
the objective is to provide a production medium that allows
optimal growth of the microorganism.
3 Continued…
 For secondary metabolite production, such as antibiotics, their
biosynthesis is not growth related.
 Consequently, for this purpose, media are designed to provide an initial
period of cell growth, followed by conditions optimized for secondary
metabolite production.
 At this point the supply of one or more nutrients (carbon, phosphorus or
nitrogen source) may be limited and rapid growth ceases.
4 FERMENTATION MEDIA
COMPONENTS

CARBON SOURCE

NITROGEN SOURCE

WATER

MINERAL

VITAMINES AND GROWTH FACTOR

NUTRIENT RECYCLE

BUFFERS

PRECURSORS

INHIBITORS

INDUCERS AND ELICITORS

OXYGEN

ANTIFOAM
5 MEDIA ADOPTED ALSO DEPEND ON
THE SCALE OF THE FERMENTATION
 SMALL SCALE FERMENTATION
 For small-scale laboratory fermentations pure chemicals are often
used in well-defined media.

 INDUSTRIAL SCALE FERMENTATION

 However, this is not possible for most industrial-scale fermentation
processes, simply due to cost, as media components may account
for up to 60-80% of process expenditure.
6 FACTORS AFFECTING THE FINAL
CHOICE OF INDIVIDUAL RAW
MATERIAL
 Cost and availability
 Ease of handling in solid or liquid forms
 Media making and Sterilization
 Formulation, mixing, complexing and viscosity characteristics that
may influence agitation, aeration and foaming during fermentation
and downstream processing stages.
 The concentration of target product
 undesired products formation
 Overall health and safety implications
7 MINIMAL QUANTITIES OF NUTRIENTS

carbon and energy source + nitrogen source + O ₂ +

other requirements →
biomass - products +CO₂ + H₂O + heat
8 NUTRIENT RECYCLE
Nutrient reuse is the process of converting municipal waste water or
industrial nutrient into nutrients that can be reused for a variety of
purposes.
The reuse or efficient use of water is normally of high priority.
ICI plc(Imperial Chemical Industries)-
When ICI plc`s and John Brown Engineering developed a continuous-
culture single cell protein (SCP) process at a production scale of 60,000
tons/ year.
After this production it was realized that very high costs would be
incurred if fresh purified water was used on a once-through basis, since
operating at a cell concentration of 30 g biomass (dw) dm would require
2700 × 10 dm of water per annum.
9 Continued…
 After that scientist carry out Laboratory tests to simulate the
process showed that the Methylophilus methylotrophus could be
grown successfully with 86% continuous recycling of supernatant
with additions to make up depleted nutrients.
 This approach was therefore adopted in the full scale process to
reduce capital and operating costs and it was estimated that water
used on a once through basis without any recycling would have
increased water costs by 50% and effluent treatment costs 10-fold.
10 BUFFERS :

 DEFINITION

Buffers are compound or mixture

of compounds that by their presence
in the solution resist changes in the
pH upon the addition of small quantities
.of acid or alkali.
11 Necessity of a buffer system:

 Sometimes it is necessary that a solution of a definite pH be

prepared and stored. The preservation of such a solution is even
more difficult than its preparation. If solution comes in contact with
air, it will absorb CO₂ and becomes acidic. On the other hand, if
solution is stored in a glass bottle, alkaline impurities from the glass
may alter its pH. Due to these reasons, pharmaceutical solutions are
buffered as the buffer solutions are capable of maintaining pH at
some fairly constant value when even small amounts of acid or base
are added.
 Continued…
12
 Many media are buffered at about pH 7.0 by the incorporation of
calcium carbonate (as chalk).
 If the pH decreases the carbonate is decomposed.
 Obviously, phosphates which are part of many media also play an
important role in buffering.
 However, high phosphate concentrations are critical in the
production of many secondary metabolites.
 The balanced use of the carbon and nitrogen sources will also form
a basis for pH control as buffering capacity can be provided by the
proteins, peptides, and amino acids, such as in corn-steep liquor.
 The pH may also be controlled externally by the addition of
ammonia or sodium hydroxide and sulfuric acid.
 TYPES OF BUFFERS :
13 1. ACIDIC BUFFER
2. BASIC BUFFER

BUFFER ACTION :
The resistance of a buffer solution
toa change in pH is known as buffer
action.
14 APPLICATION OF BUFFERS :

 PHARMACEUTICAL INDUSTRIES
 IN FERMENTATION
 FOOD INDUSTRY
 DYEING INDUSTRY
 LEATHER INDUSTRY
 OTHER MISCELLANEOUS ..
15 IN FERMENTATION :
 Fermentation reactions such as in beer or yogurt are highly affected
by varying pH.
 This means it's essential to use buffer solutions to avoid harsh
changes and allow fermentation to progress to obtain maximum
yield.
 During fermentation of baking bread the pH of the dough will
decrease due to released carbon dioxide and other organic acids. In
dough, flour and milk act as buffering agents and they resist the pH
drop due to the release of carbon dioxide.
 Apart from these natural buffers some chemical buffers like calcium
carbonate are also used to maintain pH during fermentation process.
16 REFERENCES

 Stanbury P. F., Whitaker A. and Hall S. J., (2016), Principles of

Fermentation Technology, 3th Edition, , Elsevier, (ISBN: 978-0-08-
099953-1)
 Waites, M.J., (2001) Industrial microbiology: An Introduction, 1st
Edition, Blackwell publishing, (ISBN: 0-632-05307-0)
 Google Wikipedia