Ho Chi Minh City University of Technology and Education

Faculty of Chemical and Food Technology

Department of Food Technology
Program for High Quality Training

PRINCIPLE OF FOOD PROCESSING

AND FOOD PROCESSING EQUIPMENT
(PFPE120350E – 2 credits)

Lecturer: Dang Dinh Khoi, Ph.D.

Email: [email protected]
Microbial involvement in
Food processing
Fermentation
• Fermentation is one of vital processes in living systems. As biochemical definition,
fermentation is a metabolic process which occurs when the electron transport chain
is unusable (often due to the lacking of electron receptor, such as oxygen) to
produce energy (as ATP) instead of respiration.
• In other words, fermentation is a metabolism in which organic substances (mostly
sugars) were transformed to other molecules, such as alcohol, acids, gas (CO2), or
other secondary metabolites by microorganism activities.
Fermentation
In food processing, fermentation is important because both food and
spoilage can be caused by fermentation.
Fermentation
In food processing, fermentation is one of the most traditional
methods.

Making bread and drinking beer in ancient Egypt

Fermentation
Typical events of Fermentation Time (AD)

Wine production promoted by Roman Emperor 3rd century

Fermentation of yeast by Erxleben 1818

Lactic acid and beer fermentation by Pasteur 1860s

Fermentation of enzymes in yeast by Buchner 1897

Discovery of penicillin by Alexander Fleming 1929

Discovery of other antibiotics and metabolites 1945 - present

History of events in fermentation

(Anthony Pometto, Kalidas Shetty, Gopinadhan Paliyath, 2006)
 Fermentation
Advantages and Disadvantages of fermentation
Advantages Disadvantages
• Preserving foods and beverages, • Fermentation is vulnerable to
enhancing flavors. contamination and spoilage.
• Increasing number of probiotics, • It requires equipment and
improving the health of techniques to avoid
intestinal tracts. contamination.
• Improving cardiovascular health • High acidity in fermented
and performance of immune foods may caused
system. stomachache.
Fermentation
• Two types of fermentation:

Solid-state Submerged
fermentation fermentation

Very high
Low moisture
moisture
content
Rice bran content

Solid Fermentation
substrates in liquid Submerged
fermentation
Rice husk
Fermentation
• Two types of fermentation:

Solid-state Submerged
fermentation fermentation

Limiting the Vulnerable to

contamination contamination

Hard to Easy to
harvest and
harvest apply in
products industry
Fermentation
Solid-state fermentation
• Solid state or solid substrate fermentation (SSF) involves the growth
of microorganisms (mainly fungi) on moist solid materials that have
a large surface area per unit volume (typically 103 - 106 m2/cm3).
• Traditional SSF foods include koji, Indonesian tempeh, Japanese
sake, Vietnamese “tương bần”…
• SSF is also used to produce enzymes (e.g. amylases,
amyloglucosidase, cellulases, proteases, pectinases, xylanases and
glucoamylases), organic acids, aroma compounds and flavours and
for edible mushroom cultivation.
• Substrates include rice, tubers, wheat, millet, barley, beans, maize
(corn), sugar beet pulps, soy beans, and also food processing
residues (e.g. wheat bran and soy flakes remaining after oil
extraction).
Fermentation
Solid-state fermentation
• The stages in an SSF are first to prepare a substrate by
homogenisation, enzymatic hydrolysis, grinding or flaking, and
sometimes heating to soften the material and remove
contaminants.
• It is then inoculated and incubated with the microorganism.
• Finally the required extracts are either collected or leached from the
fermented solids
Fermentation
Solid-state fermentation

Production of pectinase by using Aspergillus niger in SSF

Fermentation
Solid-state fermentation
• For microorganisms to grow on a solid substrate they should be able
to tolerate lower water activities, penetrate the substrate and utilise
complex mixtures of different polysaccharides.
• Filamentous fungi meet these requirements and their hyphae grow
into solid substrates and secrete enzymes that enable the utilisation
of available nutrients at low water activity and high osmotic
pressure/nutrient concentration.

Aspergillus niger
Fermentation
Solid-state fermentation
• Bacteria (Bacillus spp.) are used for enzyme production, and for
producing fermented products such as sausages, Japanese natto and
fermented soybean paste.
• Yeasts are used for fermentation of cassava and rice, ethanol
production and protein enrichment of agricultural residues for feeds.

Bacillus spp. Yeast

Fermentation
Solid-state fermentation
The factors that control the fermentation are:
1. Temperature;
2. Rates of gas and nutrient diffusion;
3. Water activity of the substrate.
Fermentation
Solid-state fermentation
Temperature
• Metabolic activity increases the temperature in SSFs: the extent
depends on the type of microorganism, and the porosity, particle size,
moisture content and depth of the substrate bed.
• If heat removal is inadequate, it affects spore germination, cell growth
and product formation.
• Temperature control is more difficult than in submerged
fermentations and relies on mechanical agitation or forced aeration to
transfer heat from the substrate to air and remove the heated air
Fermentation
Solid-state fermentation
Aeration
Aeration has two other functions: to supply oxygen for aerobic
metabolism and to remove CO2 and volatiles produced by cell
metabolism.
Moisture content
The water activity of the substrate affects biomass growth, metabolic
reactions and mass transfer processes. Depending on the application,
the optimum substrate moisture content (usually between 30% and
75%) allows maximum cell growth or production of metabolites.
Fermentation
Submerged fermentation
• There are a number of different designs of fermenter that are used for
submerged cultures using liquid substrates, including stirred tank,
bubble column, air-lift and tower fermenters…
• Stirred tank fermenters are widely used and have stainless steel tanks
that are fitted with cleaning-in-place (CIP) and sterilising-in-place (SIP)
facilities.
• Batch and continuous fermenters have similar controls but continuous
fermenters have additional arrangements to continually collect the
products and remove the residues.
Fermentation
Submerged fermentation
• In operation, the inoculum is produced from a freeze-dried stock
culture and introduced into a fermentation medium in a sterilised
fermentation vessel.
• The medium contains a carbon source (e.g. glucose syrup) with added
nutrients depending on the requirements of the microorganism for
growth and/or metabolite or enzyme production.
• In aerobic fermentations, air is pumped into the fermenter to initiate
a period of growth under constant conditions of temperature,
dissolved oxygen (DO) and pH, which are maintained by automatic
control mechanisms.
 Fermentation
Submerged fermentation

• Batch fermenter showing controls and

instrumentation.
• S = steam sterilising points.
Fermentation
Submerged fermentation
• In batch operation, the fermenter is emptied at the end of the
fermentation, judged by accumulated biomass and/or enzymes or
metabolites, or by depletion of nutrients.
• In continuous fermentations, a feed of nutrients is maintained with
the simultaneous removal of fermenter broth so that an excess of
nutrients remains in the supernatant.
• In both batch and continuous fermentations the main factors that
require control are temperature, pH, dissolved oxygen, degree of
agitation and foaming.
Fermentation
Submerged fermentation
Temperature control
There are three ways to control the temperature for optimum cell
growth and/or metabolite production:
1. The growth medium is continuously recirculated through a heat
exchanger;
2. The fermentation vessel is fitted with a water-jacket and water is
recirculated through the jacket and a heat exchanger;
3. The fermenter is fitted with an electric heating blanket, with each
type linked to a thermostatic controller.
Fermentation
Submerged fermentation
pH control
• A digital pH probe is linked to a controller that measures and controls
pH to an accuracy of 60.01 pH units.
• The microprocessor controls both the calibration of the pH probe
using standardised buffers and pumps to add acid or alkali to keep the
pH within preset limits.
• Electronic timers determine the length of time that pumps operate to
provide a delay between corrective additions, thus minimising
overshooting.
Fermentation
Submerged fermentation
Dissolved oxygen control
• A digital DO probe continuously monitors the DO concentration in the
medium. It is calibrated by the software in the controller with
temperature compensation, and a display indicates DO concentration
in a range of 0 - 100%.
• The required DO concentration range is maintained using upper and
lower set points, and the controller is linked to gas flowmeters and
the agitator speed monitor.
• It controls air pumps or regulators on pressurised gas tanks to add
oxygen, air, carbon dioxide or nitrogen to the vessel, and/or to control
the speed of agitation, to maintain the required DO concentration.
Fermentation
Submerged fermentation
Agitation control
A controller equipped with digital speed measurement monitors the
degree of agitation and adjusts a variable-speed motor on the mixer
shaft, typically operating between 50 and 1250 rpm.
Foaming control
Agitation and aeration may result in excessive foaming, depending on
the growth medium and the types of metabolites produced by the
fermentation.
Foaming is controlled to prevent foam being forced out of the probe
ports, thereby risking contamination; and to prevent it inhibiting
oxygen transfer.
Fermentation
Submerged fermentation
Foaming control
• Foam-sensing probes are mounted within the fermentation vessel and
when foam contacts the probes a controller drives a pump to
introduce antifoam.
• Antifoam chemicals used in food fermentations include silicone,
mineral oils, polyols, fatty acids, alcohols, hydrophobic silica and
polyalkylenes.
• Flowmeters are linked to the controller and adjustable electronic
timers determine the length of the antifoam addition and the period
of delay between doses.
Fermentation
Submerged fermentation
Foaming control
• Foam-sensing probes are mounted within the fermentation vessel and
when foam contacts the probes a controller drives a pump to
introduce antifoam.
• Antifoam chemicals used in food fermentations include silicone,
mineral oils, polyols, fatty acids, alcohols, hydrophobic silica and
polyalkylenes.
• Flowmeters are linked to the controller and adjustable electronic
timers determine the length of the antifoam addition and the period
of delay between doses.
Fermentation
Submerged fermentation
Time control
• The duration of the fermentation is controlled using a programmable
timer with automatic shutdown of the fermenter at the end of the
fermentation period.
Displays and data logging
• Data logging and control software enable the fermentation cycle to be
programmed from a computer or touchscreen, and to save different
fermentation cycle programmes.
Fermentation
Submerged fermentation

Fermenter control screen

Microbial enzymes
• Microbial enzymes are enzymes produced by microorganisms,
including molds, yeast and bacteria.
• Advances in biotechnology have had a significant effect on the
numbers and types of enzymes that are available for use in processing
and there are currently (in 2016) 250 enzymes that are commercially
available.
• Applications include bakery products, fruit juices, starch processing,
glucose syrups, dairy products and meat products…
Microbial enzymes
• Microbial enzymes have optimum activity under similar conditions to
those that permit optimum cell growth.
• They are either secreted by the cells into the surrounding medium
(“extracellular” production) or retained within the cell (“intracellular”
enzymes).
• Enzymes are separated and purified from microbial cells or
fermentation liquor, and are added to foods as either concentrated
solutions or powders.
Microbial enzymes
Extracellular enzyme production occurs in either the logarithmic phase
or the stationary phase of growth, whereas intracellular enzymes are
produced during logarithmic growth but are only released into the
medium when cells undergo lysis in the stationary or decline phases.
Microbial enzymes
Advantages of using microbial enzymes:
• Enzymic reactions are carried out under mild conditions of
temperature and pH, and are highly specific, thus reducing the
number of side reactions and byproducts;
• Enzymes are active at low concentrations and the rates of reaction are
easily controlled by adjustment of incubation conditions;
• There is minimal loss of nutritional quality at the moderate
temperatures employed;
• Lower energy consumption than corresponding chemical reactions;
• They enable the production of new foods, not achievable by other
method;
• They are friendly to the environment.
Microbial enzymes
Disadvantages of using microbial enzymes:
• The cost of many enzymes is high and, in some products, enzymes
must be inactivated or removed after processing, which adds to the
cost of the product;
• Enzymes are mostly proteins and may cause allergic responses in
some people;
• The activity of enzymes may declined quickly if they are not
adequately preserved or transported;
• Enzymes are highly specific, so it needs to use various kinds of
enzymes in the processing of foods.
Microbial enzymes
The requirements of commercial enzyme production from
microorganisms are as follows:
• Microorganisms must grow well on an inexpensive substrates;
• Substrates should be readily available in adequate quantities, with a
uniform quality;
• Microorganisms should produce a constant high yield of enzyme in a
short time;
• Methods for enzyme recovery should be simple and inexpensive;
• The enzyme preparation should be stable.
Microbial enzymes
• Enzymes are produced by either solid substrate fermentations (e.g.
rice hulls, fruit peels, soy bean meal or wheat flour) or by submerged
culture using liquid substrates in fermenters (e.g. molasses, starch
hydrolysate or corn steep liquor).
• Submerged cultures have lower handling costs and a lower risk of
contamination and are more suited to automation than are solid
substrates.
• Extracellular enzymes are recovered from the fermentation medium
by centrifugation, filtration or membrane separation, fractional
precipitation, chromatographic separation, electrophoresis, freeze
drying, or a combination of these methods.
Microbial enzymes
• Intracellular enzymes are extracted by disruption of cells in a
homogeniser or mill.
• Recovery is more difficult and the yield is lower than for extracellular
enzymes, because some enzymes are retained within the cell mass.
• If required, the specific activity of the enzyme is increased by
precipitation using acetone, alcohols or ammonium sulphate or by
ultrafiltration.
Microbial enzymes
• In batch operation, the enzyme is mixed with food, allowed to catalyse
the required reaction, and then either retained within the food or
inactivated by heat.
• This method is used when the cost of the enzyme is low. In continuous
operation, immobilised enzymes are either mixed with a liquid
substrate and removed by centrifugation or filtration and re-used, or
the feed liquor is passed over an immobilised bed of enzyme in a
bioreactor.
• Immobilisation is used when an enzyme is difficult to isolate or
expensive to prepare.