FERMENTATION

FERMENTATION
 The oldest form of food preservation.
 Encourage multiplication of m/o.
 Definition
– Chemical conversion of carbohydrates into
alcohol or acids

Sugar yeast Alcohol/acid/CO2

Cont.
 Zymology – science of fermentation
 Ethanol fermentation product – wine, beer, bread
 Lactic acid fermentation product – sour food
(yogurt,kimchi)– also by mamalian muscle

 Microbes
– Alter the texture of foods
– Preserve foods (product of alcohol/ acid)
– Produce subtle flavours and aromas

 Supplemented with other operation

– Pasteurization
– Chilling
– MAP
 Lactobacillus lactis – dairy
 Steptococcus thermophilus –dairy
 Leuconostoc sp. – wine, dairy
 Pediococcus sp. – meat, vegetable,
ripening of some cheeses
 Lactobacillus sp. – meat, vegetable, dairy,
sourdough
 Bifidobacterium sp. – dairy – promote
intestinal health
 Propiopnibacterium sp. – Swiss cheese
 Yeast – bread, beer, wine, liquors
 Molds – ripening cheeses, soy sauce
 Lactobacillus delbruekii,subspecies
bulgaricus & Streptococcus thermophilus –
yogurt
 Benefits !
 Preservation – inhibitory to the common pathogenic
m/o
(Clostridium botuinum)-can’t grow at
pH<4.6.

 More nutritious – not only catabolic, but also

anabolic  synthesize complex of
vitamin (riboflavin, B12,C) protein

– mold  rich in cellulose-splitting

mycilea
enzyme

 Disease prevention – Paracoccus denitrificans (carrot

 Factors Controlling Fermentation
 Acid
– Have inhibitory effect
– Naturally formed/added to prevent further
spoilage
– In acid cond, preservative power increased. If
O2 available – oxidation occurred – natural
cond.
– Eq: milk – naturally fermented cheese

 Alcohol
– Preservative effect
– Depends on concentration (12-15% by volume)
– Sometimes need pasteurization (increase
 Starter
– Selected strains of lactic acid organism
– Quick & dependable acid production
– Specified culture are available
– Before put the starter, heated-- inactivate
other competitors
– Ancients times – naturally created

 Temperature
– Depends on the ability of m/o. temp, acid
content.
– <37°C, more bacteria produce acid-- kill other
m/o.
 O2
– Aerobic condition encourage development of
mold
– Mold oxidize acetic acid to CO2 & water

 Salt
– Added to control/inhibit proteolytic organism &
spoilage types.
– Frequently added as water drawn from vege
will dilute brine solution.
 Sauerkraut – 2.0-2.5% added into cabbage
 Pickled olive – 7-10% added into olive
 Pickled cucumber – 15-18% added
 Eskimos
– Prepared fermented fish by digging a
hole about 2 ft in the ground.
– Put the freshly caught fish in the hole
– Covers with earth & let it buried for a
couple of weeks/> 1 month.
– After reaching the desired level of
fermentation – unearth – freeze
immediately eat
– Tastes best raw and frozen
 African
– Fermented fish popular in Ghana,
Uganda, Burundi.
– However unclean practices may cause
dangerous consequences.
– Factors
 Water
 Cleaning practices
 Gutting
 Environment
 Fish Sauce
 Salty, smelly brown liquid made from
fish that is the single, most important
flavoring ingredient in Thai cooking
(Laos, Cambodia, Vietnam, Burma &
Philipines).
 Called “nam bplah” in Thai, genuine
fish sauce is the water, or juice, in
the fish that is extracted in the
process of prolonged salting &
fermentation.
 Processing
Small fish

Rinsed & drained

Mixed with sea salt

Filled into large earthenware jars

On the bottom with a layer of salt topped with a layer

of salt

Covered & left in a sunny location for 9 months-year

* Protein content increase (9.6 to 15.2%)-from

 SOY SAUCE
MANUFACTURING
 INTRODUCTION
SOY SAUCE (shoyu)
THIN SAUCE THICK SAUCE
 Salt > 14% (salty)  Salt ( 10 – 13%)
 Contains more sugar,
caramel, conditioner, flavouring
Fermentation 40 days – 6 Fermentation 6 months – 24
months months.
 cheaper  Expensive than thin sauce.

Under the same legislation : Regulation 340 & Regulation

28 (Food Act 1983 & Food Regulation 1985)
 INGREDIENTS
 Defatted soy bean/mature
soy bean
 Wheat flour
 Innoculum (culture of
Aspergillus oryzae)
 Brine solution
(conc.:depends on types of
soy sauce)
 Sugar
 Caramel
 Food preservatives (eg:
benzoic acid)
 Flavour enhancer (eg:
 MANUFACTURING METHOD
SOY SAUCE

Chemical and microbiological

Chemical Method
Method
- Mineral acid – hydrolyse - Acid hydrolysed and
the protein & carbohydrate fermentation to produced
shinshiki
Microbiological Method

- 2 step fermentation
 PROCESSING OF SOY SAUCE

 Traditional method

 Modern method
 produce high Q of soy sauce
 fulfill the requirement in Food
Regulation 1985.
 MODERN METHOD
washing
Soaking Boiling
Soy bean Cooling Mixing
(10 hrs) (20min,15psi) (Wheat Flour,20%
*remove seed Innoculum,0.1%)
coats/hulls *horizontal retort *Aspergillus spp.
Incubate in
Put In Koji Put onto
Koji Room
(greenish rack Put In Tray-2cm thick
Fiber Tank (30°C,RH 90%,
yellow) (stainless steel)
48 hrs)
+ brine(22%)
[1:3-koji wt :brine wt]
Moromi
*sampling for Kjeldahl
Fermentation method-not less than
2-18 months 0.9%N2

Raw Soy Aging Tank Sedimentation

Extraction Filter
Sauce (1 week – sunlight) Tank
& Filtration

Pasteurization Mixing
Bottling (80°C,30min) Tank
PROCESSING PLANT
 CRITICAL CONTROL POINT
STEP HAZARD

1. Ingredient of  moisture
koji; soy bean, Insect
wheat flour. Microorganisms

2. Boiling of soy Uncooked soy bean: effect growth of mould & N2

bean production
3. Innoculum  low Q of innoculum – effect fermentation of koji.
4. Koji  uncontrolled temp:-
- high temp - inhibit the growth of A. oryzae
- low temp. – growth of Rhizopus nigrificans (black
mould)
*Best temp. in fermentation of koji
30°C - 35°C.
 uncontrolled moisture

- high moisture – growth of bacteria

* Best moisture content of soy bean
43% - initial of incubation
 CRITICAL CONTROL POINT….cont.

STEP HAZARD
5. Concentration of brine  less 20% - enhance
the growth of undesired
microorganism

6. Pasteurization  incomplete – effect the

flavour and the shelf life
of soy sauce.

7. Cleaning and  unsanitized of the

sanitizations factory will effect the
whole of soy sauce
production.
 CURRENT ISSUE
 Before the Gazette of Food Regulation 1985, a lot of soy
sauce manufacturers:

 Used saccharin as sweetener in soy sauce.

 Used recycled alcoholic bottles.
 Addition of Acid Hydrolysed Vegetable protein to accelerate
production of soy sauce.

 Problem in meeting the requirements of the total

nitrogen of soy sauce – because of the poor processing

3. The main source of 3-MCPD (3-monochloropropane-1,2-

diol) in soy sauces is from the addition of acid-hydrolysed
vegetable protein (acid-HVP), a savoury food ingredient
which has been found to contain 3-MCPD.
4. 3-MCDP is a group of chemicals known as
chloropropanols.
 MAKING OF KOJI
 After boiling, the soy bean will be
cooled and mixed with wheat flour
and spreaded on the bamboo tray –
incubate for 3 – 4 days for allowing
the growth of selected mould
(Aspergillus spp.) KOJI formed

 No deliberate innoculation repeated

 Use of same trays (ensure that same
organism are carried from one
fermentation batch to another)
 FERMENTATION
 Koji and brine (16 –
18%) are put in
earthen jar under sunlight
(10 – 12 weeks).
 Cover during the night
& rainy day
 Moromi formed
 SORTING
 Good Q of soy bean is
important in making soy
sauce.
 During sorting – defected
soy beans are separated.
 BOILING
 Soy beans are boiled in
a boiler (horizontal retort
(20 min, 15 psi).
 Boiling – breakdown
the protein structure of
the soy beans – *will be
hydrolyzed by the
protease (Aspergillus
oryzae).
MIXING
 Cooled soy beans +
wheat flour (20%) +
innoculum of
Aspergillus oryzae
(0.1%) in mixer, eg:
ribbon mixer.

Aspergillus oryzae
STAGE 1 :KOJI
 The mixture is spread (2cm
thick) onto the stainless
steel tray (with hole).
 These trays are covered
with muslin cloth.
 Incubate in koji room–
30°C/48 hrs/RH 90%
 At this stage – protein,
oligo and polysaccharides
are hydrolyzed –
Aspergillus enzyme
 This results in production
of fermentable sugar,
peptides and amino acids.
 Mould grow Koji formed
(greenish yellow - good Q).
 Longer incubation – allow
the growth of undesirable
black moulds.
FERMENTATION
 Koji - put into food grade fiber
tank – equipped with cover,
valve, pipe, filter, air flow.
 Brine (22%) is put into this
tank. 1:3 (koji’s wt : brine’s wt)
– to kill the growth of
Aspergillus oryzae.  moromi
formed
 Lactic fermentation occurs
– Lactobacillus delbruckii &
Pediacoccus soyae
 Followed by alcoholic
fermentation (by yeast) –
Zygosaccharomyces rouxii
 Exposed to the sunlight –
Bright and sunny weather
tends to be more conducive in
producing soy sauce of better
Q and aroma.
 Exposure for 2 months – to get
the min aroma of soy sauce
and continuously developed.
 To ensure the conc. of
the brine not less than
20% - use brinometer.

 Fermentation of
moromi within 45 days
(min period to dissolve
56%- 68% of total Brinometer
nitrogen from soy
bean).

 Total nitrogen in raw

soy sauce (45 days) -
0.9 – 1.1% - depends
on the type of
fermentation tank ; Moromi
fiber/earthen
jar/stainless steel.


 EXTRACTION & FILTRATION
 After fermentation, raw soy sauce
will be extracted and filtrated to
aging tank – using hydraulic/pump.
 In aging tank – the mixture sunned
for a week– to develop aroma and
flavour.
 SEDIMENTATIONS
 The raw soy sauce will be pumped to
the sedimentation tank.
 Sampling for the total nitrogen using
Kjeldahl method – the total nitrogen
supposed to at least 0.9% before
mixing with other ingredients and
pasteurization – may lost during
heating.
 The processed soy sauce must
contains at least 0.6% total nitrogen
MIXING
 Raw soy sauce with
sufficient nitrogen will be
pumped to the mixing
tank.
 Brine (18%), sugar,
caramel and food additive
will be added in the ratio
1:2 (vol ingredient : vol
raw soy sauce).
 Caramel and sugar  to
condense and sweeten the
sauce.
 Caramel  calouring agent
 Food preservative ;
eg : benzoic acid – not
more than 1000 ppm (Food
Regulation 1985).
 Flavour enhancer also can
 PASTEURIZATION
 Raw soy sauce will be
pasteurized at 80°C/30
min in mixing tank
equipped with
 stirrer and
jacketed thermal
oil (to prevent –
burnt and crusty),
or
 Plate heat
exchanger.
BOTTLING
 The processed soy
sauce will be filled
into sterilized
bottles using high
pressure vacuum.
 Soy sauce –
capped, labelled
and distributed.
 Conclusion
 Allof the food potential to be
fermented (naturally/ modified), but
if there’s no controlled handling,
proteolitic and lipolytic will replace
the process and spoiled the food.
 TODAY’S THORN

IS TOMORROW’S FLOWER
 CHARACTERISTICS OF FIBRE
TANK
 Food grade
 recyclable and allows for easy
disposal
 high-temperature performance
 corrosion resistance