FE 310 FOOD TECHNOLOGY

3 WEEK
RD

THERMAL PROCESSING

Prof. Dr. Ahmet Yemenicioğlu

 THERMAL RESISTANCES OF MICROORGANISMS
Thermal Resistance of Virus
While a few minutes at 50-70oC is sufficient to
denature the proteins in the viral capsid (i.e.,
protein membrane enveloping the viral DNA or RNA),
the nucleic acids can remain infectious when extracted
from the denatured virus. In order to inactivate the
viral nucleic acids, a higher input of heat is necessary.
The actual thermal resistance of virus in canned foods
is not relative since most that have public health
significance, such as hapatitis A virus, are denatured in
at least 10 minutes in moist heat at 100oC, a process
that is exceeded in almost all present-day thermal
processes for low-acid canned foods. 2
 THERMAL RESISTANCES OF MICROORGANISMS
Thermal Resistance of bacteria

Those of most concern to canners form spores when vegetative growth has
been restricted and can survive several hours of moist heat at 100oC

(For example it takes almost 22min to inactivate 90 % of C. sporogenes-3679

spores at 116oC).

However, only a minority of the bacterial species are capable of forming

spores, which is a cellule that is more resistant than the vegetative form.

Sporulation permits survival of the organism for extended periods in a

physicochemical environment not conducive to vegetative growth.

When the availability of carbon, nitrogen, and phosphorus necessary for

growth is limited, many species of bacteria (e.g., aerobic Bacillus, the
anaerobic Clostridium, as well as a few Sporosarcina and Actinomycetes)
produce dehydrated elements, or endospores that contain the essential
cellular components.

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 THERMAL RESISTANCES OF MICROORGANISMS
Thermal Resistance of bacteria

Bacterial ENDOSPORES show no metabolic activity and

are well adapted for prolonged survival under adverse
conditions.

The SPORE WALL made up of normal peptidoglycan provides an

irreversible osmotic stabilization to the endospore and the
elements necessary for the germination.

The CORTEX a less cross-linked peptidoglycan represents 15% of the

spore mass.

The very thick SPORE COAT is rich in keratin, which shows a number
of disulfide bridges and represents 80% of the total protein content
of the spores.

Spore germination is initiated under the influence of a stress such

as heat shock, pH deviation, -SH compounds.
ENDOSPORE
 THERMAL RESISTANCES OF MICROORGANISMS
Thermal Resistance of bacteria

Bacterial ENDOSPORES also contain EXOSPORIUM the most

ENDOSPORE
external coat that surround the endospore. The exospotium
is formed by the mother cell. The exposporium gives
endospore resistance against chemical disinfectants.
THERMAL RESISTANCES OF MICROORGANISMS

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 THERMAL RESISTANCES OF TOXINS
Toxins
Most toxins are globular proteins. If you denaturate them by heating,
they loose their activity.
Botulinal toxins are heat stable. The toxin which can be destroyed
by exposing moist heat for 10 min at 100oC is extremely potent (one
millionth of a gram may kill man).
Staphylococcal toxins (protein) and mycotoxins (not a protein, but
an organic compound) produced by molds are also extremely heat
stable. Especially, aflatoxins of Aspergillus flavus and A. parasiticus
are very heat stable and they can survive from canning. Aflatoxin B1
may survive from deep frying, but it can be destroyed at 267oC.
Except for patulin, the mycotoxins can not diffuse from infected to
healthy tissues. Thus, peeling or removal of affected tissue controls
Staphylococcal
mycotoxin level. enterotoxin Aflatoxin B1
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 THERMAL RESISTANCES OF ENZYMES
Enzymes
Most enzymes are inactivated by canning.
However, mold polygalacturonase which may be
found in infected fruits may survive from canning
and cause softening of canned fruits.
Sometimes we observe active peroxidases (POX)
in canned food. Why?
Answer:
1.Some extremely thermostable plant POX may
survive from canning.
2.After complete thermal inactivation some
peroxidase (not all) might show renaturation (in past
called wrongly regeneration) and regain its activity.
However, in general, most peroxidases are
inactivated irreversibly during blanching. 8
 THERMAL RESISTANCES OF PRIONS
Prions
-Transmissible Spongiform Encephalopathies (TSE) are the best known prion
diseases. The TSE cause fatal neurodegenerative disorder.
-Two diseases caused by prions are; 1.BSA mad cow disease observed in 1987 in
GB and 2.CJD (Creutzfeldt-Jacob Disease), the best known human prion
diseases
-Usual food processing parameters are largely unable to kill prions.
The Fo (the heating times at 121.1oC) desired to inavtivate prions ranges
between 200-600min, but in food industry the Fo is between 20-60 min.
-The US Department of Health and Social Security (DHSS 1984) suggested
18min at 134 oC (± 4 oC) for sterilisation of CJD infected material.
-The American Neurological Association (ANA 1986) suggested 60min at
132oC. 9
 DEATH OF BACTERIA SUBJECTED TO MOIST HEAT
The death of bacteria subjected to moist heat follows a first order reaction kinetic and can be expressed by the following
equation;
– dC / dt = k C
C= concentration of decomposing reactant; k= rate constant; – dC / dt= rate at which concentration decreases
– dC / C = k dt

𝐶𝐶𝐶 𝑡𝑡2
- ∫𝐶𝐶1 dC / C = k ∫𝑡𝑡1 dt
– ln C2 – (–ln C1)= k (t2 – t1)
k = ln C1 – ln C2 / (t2 – t1)= [2.303 / (t2 – t1)] log C1 / C2
k= [2.303 / t] log Co / C
Co= initial conc. (No= initial number of m.o.); C= conc. after t time (N= number of m.o. after t time)
k= [2.303 / t] log No / N
k= [1 / t] ln No / N
ln N / No = – k t 10
CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)

D value (Decimal reduction time)

Time at a constant temperature,
required to kill 90% of the organisms
present or time at a constant
temperature, required to obtain one
decimal (log) reduction in the number
of m.o. The D value can be calculated
from heat inactivation curve (thermal
inactivation curve, survivor curve)
which obtained by plotting number of
survivals versus heating time on semi-
log scale.
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CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)

Tan α= log 100 – log 10 / tb-c

Tan α= slope = 1 / D
D=1 / Slope

k= [2.303 / t] log No / N
k= [2.303 / t] log 100 / 10
k= 2.303 / D= Slope x 2.303

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CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)

Analysis of heat inactivation curve

(Equil triangles)

ABC ~ abc
AB / ab = BC / bc
log No – log Nt / log 100 – log 10 = t / D
log No / N = t / D
log N / No = -t / D
D= t / log No – log N

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CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)

PROBLEM 1
The initial number of PA-3679 is 104 spore/tube. The number of
spores reduced to 90 spore/tube after 10min heating at 240 oF
constant temperature. Calculate D value of the microorganism.

D= 10 / log 104 – log 90

= 4.88 min
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 CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)
PROBLEM 2
The remaining activities of peroxidase (POD) enzyme, determined after various heating times at 70o, 75o and 80oC at
pH 7.0 phosphate buffer were given below. Calculate the D values of the enzyme at different temperatures.

Time (min) Residual activities (%)

70oC 75oC 80oC
0 100 100 100
1 90 70 54
3 75 50 5.3
5 58 30 1.7
7 47 18
9 37 7.9
11 29 4.6
13 24 3.2
15 19 - -
17 16 - -
15
20 13 - -
 CALCULATION OF HEAT INACTIVATION PARAMETERS (1-D value)
PROBLEM 2
The remaining activities of peroxidase (POD) enzyme, determined after various heating times at 70o, 75o and 80oC at
pH 7.0 phosphate buffer were given below. Calculate the D values of the enzyme at different temperatures.
100

70oC
Remaining activity (%)

10

75oC
80oC

1
0 5 10 15 20 25
Time (min)
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 CALCULATION OF HEAT INACTIVATION PARAMETERS (2-z value)

z value: Number of degrees of Fahrenheit or Centigrade required for

thermal death time curve to traverse one logarithmic cycle.

This inactivation parameter shows the temperature dependency of heat

inactivation. High z means low temperature dependency while low z
means high temperature dependency.

The z value is calculated by using two different methods.

1. From Decimal Reduction Curves (Phantom curve)

2. From Thermal Death Time curves (TDT curves)

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 CALCULATION OF z FROM PHANTOM CURVE
Decimal Reduction Curves (Phantom curve) 1. Decimal Reduction Curves (Phantom
curve)
The phantom curve is obtained by plotting
D values versus temperatures on a semi-
log curve.

Tan α = log 10 – log 1/ z

Slope = 1 / z
z = 1 / Slope

Analysis of Phantom curve (Equil triangles)

ABC ~ abc
ab / AB = bc / BC
log 10 – log 1 / log D2 – log D1= z / T1- T2
z = T1- T2 / log D2 – log D1
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 CALCULATION OF z FROM PHANTOM CURVE
PROBLEM 3
Calculate the z value of POD enzyme given in problem 2.

100

22,00
D value

10 9,00

3,00

1
60 70 80 90
Temperature (o C)
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 CALCULATION OF z FROM PHANTOM CURVE
PROBLEM 3
Calculate the z value of POD enzyme given in problem 2.

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 CALCULATION OF z FROM THERMAL DEATH TIME CURVE (TDT CURVE)

The use of Thermal Death Time curves (TDT curves)

In this method Thermal Death Time tubes (TDT tubes; inside diameter 9mm; wall thickness 1mm) or cans containing
the same number of microbial cells or spores (or the same level of enzyme activity) are heated at different temperatures
to find the required time period to reduce the initial number of microorganisms to 100 at each temperature.

This time period is between the highest times that we observe a survivor (H) in TDT tubes and the lowest times
that we observe no survivor (L) in TDT tubes. To calculate the z value the maximum and minimum times are plotted
versus temperature. The reciprocal of the slope of best straight line drawn immediately below the death points and
immediately above the survivor points gives the z value (Log H + Log L/2=A; Antilog (A)=Average).

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 CALCULATION OF z FROM THERMAL DEATH TIME CURVE (TDT CURVE)

TDT experiments conducted by a single tube at 120oC

constant temperature

TDT at 120oC is between 4 and 5 min (~4.5 min)

Heating 0 1 2 3 4 5 6 7 8
time, min
Results + + + + + - - - -

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 CALCULATION OF z FROM THERMAL DEATH TIME CURVE (TDT CURVE)

TDT experiments conducted by 5 tube at 120oC

constant temperature

TDT at 120oC is between 3 and 6 min (average 4.5 min)

Heating 0 1 2 3 4 5 6 7 8
time, min
+ + + + + + + - -
+ + + + + + - - -
Results + + + + + - - - -
+ + + + - - - - -
+ + + - - - - - -
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CALCULATION OF z FROM THERMAL DEATH TIME CURVE (TDT CURVE)

Problem 4: Calculate the z value of the given bacterial spore from given data

Temp Log H Log L Average Antilog (A)

(oF) La Hb (A)

212 120 150 2,08 2,18 2,13 135

222 40 50 1,60 1,70 1,65 45
230 20 24 1,3 1,38 1,34 22
239 6.25 7 0,80 0,90 0,82 6.6
248 2 2.5 0,3 0,40 0,35 2.25

A Lowest time that we observe no survivors (min)

B Highest time that we observe a survivor (min)
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 CALCULATION OF z FROM TDT CURVE

Problem 4: Calculate the z value of the given bacterial spore from given data

1000,00

100,00
TDT times (min)

10,00

1,00
210,00 220,00 230,00 240,00 250,00
Temperature (o F)
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ÇOK TEŞEKKÜRLER

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