CHE 503: BIOCHEMICAL ENGINEERING

STERILIZATION, DESIGN AND

OPERATION

Lecturer:
ENGR. MICHAEL ALLAN G. RAMOS
Department of Chemical Engineering
Technological Institute of the Philippines
1st Semester, 2019-2020
 STERILIZATION
Definition
- absence of detectable levels of viable organisms in a culture
medium or gas.
- complete removal of all microorganisms.
- A requirement for an aseptic process.
- reason for sterilization: economics, loss of product due to
contamination is a costly price to pay. No chance of recovery.

Disinfection
- to reduce number of viable pathogenic microorganisms where
they can no longer infect

Pasteurization
- method of reducing the # of viable microorganism from a
product (such as milk or beverage) to extend shelf-life
 METHODS OF STERILIZATION

1. Thermal
- preferred for large-scale operation; moist heat usually
- pressurization leading to production of wet steam
- 30psia, 1210C, 15 minutes

2. Chemical
– preferred for heat-sensitive equipment
- ethylene oxide (gas) for equipment
- 70% ethanol solution (acidified to pH =2 using HCl) for surfaces of
equipment
- 3% sodium hypochlorite solution for surfaces

3. Radiation – UV for surfaces, X-ray for liquids

4. Filtration – membrane ultra-filters with 1 nm size to filter off viruses and

bacteria; for oxygen or air sterilization
 METHODS OF STERILIZATION

1. sterilization of all equipment especially inside surfaces where

contamination can start at crevices, roughness etc.

2. sterilization of the medium, so that contaminants will not

compete getting ‘food’ vs. your target or microorganism of choice

3. can be done direct or indirect. Batch or continuous

RESISTANCE OF MO’S TO MOIST HEAT
 THERMAL METHOD
1. Not all organisms have identical death kinetics.
→ (increasing difficulty; vegetative cells < spores < virus)

2. Individuals within a population of the same organism respond

differently

Probability Theory:

P(t) = the probability that an individual cell is still viable at time t.

- k d .t
P(t ) = e
(simplest form assuming 1st order death kinetics)
- 0 = will not occur; 1 = will occur
1 - Po (t ) = 1 - 1 - e [ ]
- k d .t N o probability of an
unsuccessful sterilization
 THERMAL METHOD

[
1 - Po (t ) = 1 - 1 - e ]
- k d .t N o

Po (t ) = probability of extinction of a population at time=t,

<=1
kd = death rate constant, 1/time (1st order kinetics)

t= holding time, exposure time, time (temp. dependent)

No = initial number of viable microorganism at time=0,

# of spores

Use probability chart to evaluate 1-Po(t), 0.001 is the acceptable safe

standard probability
With increments,
0.18,0.018 etc..
 Temperature Effects on Kinetics
of Thermal Sterilization

Arrhenius equation
- Ed
k d = a .e R .T

kd = Death rate constant, 1/time

a= Frequency factor, 1/time
R= Gas constant, 8.314 J/kmole-K
Ed = Deactivation energy, (50-150 kcal/g-mole = spores)
(2-20 kcal/g-mole = vitamins/ growth factors

T = Sterilization temperature, K
 Temperature Effects on Kinetics
of Thermal Sterilization

Ed
-
k d = a .e R .T

Ed 1
ln k d = ln a - . ln k d
R T
 THERMAL METHOD

dN
= - k d .N
dt
dN
= -k d .dt
N
N
ln = -k d .t
No
- k d .t
N = N o .e
N = final # of viable MO’s
 THERMAL INACTIVATION
Goal is to kill microorganisms as contaminants as well as to preserve
the chemical compounds/ nutrients in fermentation broths
 HTST
- Degradation of vitamins/ nutrients in the media can be avoided via brief
exposure to high temperature
 BATCH vs. CONTINUOUS
HEAT STERILIZATION

BATCH steam out

STERILIZATION
steam in
BATCH vs. CONTINUOUS
HEAT STERILIZATION

CONTINUOUS
STERILIZATION
Pe/ Da chart
Pe/ Re chart
END.