fermentation

FERMENTATIONFERMENTATION

FERMENTATIONFERMENTATION

The oldest form of food preservation. The oldest form of food preservation. Encourage multiplication of m/o.Encourage multiplication of m/o. DefinitionDefinition

– Chemical conversion of carbohydrates into Chemical conversion of carbohydrates into alcohol or acidsalcohol or acids

SugarSugar yeast Alcohol/acid/CO2 Alcohol/acid/CO2

Cont.Cont.

Zymology – science of fermentationZymology – science of fermentation Ethanol fermentation product – Ethanol fermentation product – winewine, , beerbeer, bread, bread Lactic acid fermentation product – sour food Lactic acid fermentation product – sour food

(yogurt,kimchi)– also by mamalian muscle(yogurt,kimchi)– also by mamalian muscle

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

Supplemented with other operation Supplemented with other operation – PasteurizationPasteurization– ChillingChilling– MAPMAP

Lactobacillus lactis Lactobacillus lactis – dairy – dairy Steptococcus thermophilusSteptococcus thermophilus –dairy –dairy Leuconostoc Leuconostoc sp. – wine, dairysp. – wine, dairy Pediococcus Pediococcus sp. – meat, vegetable, ripening sp. – meat, vegetable, ripening

of some cheesesof some cheeses Lactobacillus Lactobacillus sp. – meat, vegetable, dairy, sp. – meat, vegetable, dairy,

sourdoughsourdough Bifidobacterium Bifidobacterium sp. – dairy – promote sp. – dairy – promote

intestinal healthintestinal health Propiopnibacterium Propiopnibacterium sp. – Swiss cheesesp. – Swiss cheese Yeast – bread, beer, wine, liquorsYeast – bread, beer, wine, liquors Molds – ripening cheeses, soy sauceMolds – ripening cheeses, soy sauce Lactobacillus delbruekii,Lactobacillus delbruekii,subspecies subspecies

bulgaricusbulgaricus & & Streptococcus thermophilusStreptococcus thermophilus – – yogurtyogurt

Benefits !Benefits ! Preservation – inhibitory to the common pathogenic m/o Preservation – inhibitory to the common pathogenic m/o

((Clostridium botuinumClostridium botuinum)-can’t grow at pH<4.6.)-can’t grow at pH<4.6.

More nutritious – not only catabolic, but also More nutritious – not only catabolic, but also anabolic anabolic synthesize complex of synthesize complex of

vitamin (riboflavin, B12,C) proteinvitamin (riboflavin, B12,C) protein – – moldmold rich in cellulose-splitting enzyme rich in cellulose-splitting enzyme

Disease prevention – Disease prevention – Paracoccus denitrificans Paracoccus denitrificans (carrot (carrot juice)juice)

– – reduce reduce nitratenitrate content in body content in body Develop flavor/aromaDevelop flavor/aroma

mycilea

Factors Controlling FermentationFactors Controlling Fermentation

Acid Acid – Have inhibitory effectHave inhibitory effect– Naturally formed/added to prevent further Naturally formed/added to prevent further

spoilagespoilage– In acid cond, preservative power increased. If O2 In acid cond, preservative power increased. If O2

available – oxidation occurred – natural cond. available – oxidation occurred – natural cond. – Eq: Eq: milkmilk – naturally fermented – naturally fermented cheese cheese

AlcoholAlcohol– Preservative effectPreservative effect– Depends on concentration (12-15% by volume)Depends on concentration (12-15% by volume)– Sometimes need pasteurization (increase Sometimes need pasteurization (increase

preservation)preservation)

StarterStarter– Selected strains of lactic acid organismSelected strains of lactic acid organism– Quick & dependable acid productionQuick & dependable acid production– Specified culture are availableSpecified culture are available– Before put the starter, heated-- inactivate other Before put the starter, heated-- inactivate other

competitorscompetitors– Ancients times – naturally createdAncients times – naturally created

TemperatureTemperature– Depends on the ability of m/o. temp, acid content.Depends on the ability of m/o. temp, acid content.– <37<37°°C, more bacteria produce acid-- kill other m/o.C, more bacteria produce acid-- kill other m/o.

O2O2– Aerobic condition encourage development of Aerobic condition encourage development of

moldmold– Mold oxidize acetic acid to CO2 & waterMold oxidize acetic acid to CO2 & water

SaltSalt– Added to control/inhibit proteolytic organism & Added to control/inhibit proteolytic organism &

spoilage types.spoilage types.– Frequently added as water drawn from vege Frequently added as water drawn from vege

will dilute brine solution.will dilute brine solution. Sauerkraut – 2.0-2.5% added into cabbageSauerkraut – 2.0-2.5% added into cabbage Pickled olive – 7-10% added into olivePickled olive – 7-10% added into olive Pickled cucumber – 15-18% addedPickled cucumber – 15-18% added

Eskimos Eskimos – Prepared fermented fish by digging a Prepared fermented fish by digging a

hole about 2 ft in the ground. hole about 2 ft in the ground. – Put the freshly caught fish in the holePut the freshly caught fish in the hole– Covers with earth & let it buried for a Covers with earth & let it buried for a

couple of weeks/> 1 month.couple of weeks/> 1 month.– After reaching the desired level of After reaching the desired level of

fermentation – unearth – freeze fermentation – unearth – freeze immediately eatimmediately eat

– Tastes best raw and frozenTastes best raw and frozen

African African – Fermented fishFermented fish popular in Ghana, popular in Ghana,

Uganda, Burundi.Uganda, Burundi.– However unclean practices may cause However unclean practices may cause

dangerous consequences.dangerous consequences.– Factors Factors

WaterWater Cleaning practicesCleaning practices GuttingGutting EnvironmentEnvironment

Fish SauceFish Sauce

Salty, smelly brown liquid made from Salty, smelly brown liquid made from fish that is the single, most important fish that is the single, most important flavoring ingredient in Thai cooking flavoring ingredient in Thai cooking (Laos, Cambodia, Vietnam, Burma & (Laos, Cambodia, Vietnam, Burma & Philipines).Philipines).

Called “nam bplah” in Thai, genuine Called “nam bplah” in Thai, genuine fish sauce is the water, or juice, in the fish sauce is the water, or juice, in the fish that is extracted in the process of fish that is extracted in the process of prolonged salting & fermentation.prolonged salting & fermentation.

ProcessingProcessing

Small fishSmall fish

Rinsed & drainedRinsed & drained

Mixed with sea saltMixed with sea salt

Filled into large earthenware jarsFilled into large earthenware jars

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

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

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

SOY SAUCE SOY SAUCE MANUFACTURING MANUFACTURING

INTRODUCTIONINTRODUCTION

SOY SAUCE (shoyu)SOY SAUCE (shoyu)THIN SAUCETHIN SAUCE THICK SAUCETHICK SAUCE

Salt > 14% (salty)Salt > 14% (salty) Salt ( 10 – 13%)Salt ( 10 – 13%) Contains more sugar, Contains more sugar, caramel, conditioner, flavouringcaramel, conditioner, flavouring

Fermentation Fermentation 40 days – 6 40 days – 6 monthsmonths

Fermentation Fermentation 6 months – 6 months – 24 months.24 months.

cheapercheaper Expensive than thin sauce.Expensive than thin sauce.

Under the same legislation : Regulation 340 & Regulation 28 (Food Act 1983 & Food Regulation 1985)

INGREDIENTSINGREDIENTS Defatted soy bean/mature soy Defatted soy bean/mature soy

beanbean Wheat flourWheat flour Innoculum (culture of Innoculum (culture of

Aspergillus oryzaeAspergillus oryzae)) Brine solution (conc.:depends Brine solution (conc.:depends

on types of soy sauce)on types of soy sauce) SugarSugar CaramelCaramel Food preservatives (eg: Food preservatives (eg:

benzoic acid)benzoic acid) Flavour enhancer (eg: MSG)Flavour enhancer (eg: MSG)

MANUFACTURING METHODMANUFACTURING METHOD

SOY SAUCESOY SAUCE

Chemical Method

Microbiological Method

Chemical and microbiological Method

- Mineral acid – hydrolyse the protein & carbohydrate

- 2 step fermentation

- Acid hydrolysed and fermentation to produced shinshiki

PROCESSING OF SOY SAUCEPROCESSING OF SOY SAUCE

Traditional methodTraditional method

Modern method Modern method produce high Q of soy sauce produce high Q of soy sauce fulfill the requirement in Food fulfill the requirement in Food Regulation 1985. Regulation 1985.

MODERN METHODMODERN METHOD

CoolingBoiling

(20min,15psi)Soaking(10 hrs)

Soy bean

Put In Tray-2cm thick(stainless steel)

Koji(greenishyellow)

Mixing(Wheat Flour,20%Innoculum,0.1%)

Put In Fiber Tank

Incubate in Koji Room

(30°C,RH 90%,

48 hrs)+ brine(22%)

[1:3-koji wt :brine wt]

Moromi

Extraction& Filtration

Sedimentation Tank

Aging Tank(1 week – sunlight)

Pasteurization(80°C,30min)

*horizontal retort *Aspergillus spp.

Put onto rack

Fermentation2-18 months

*sampling for Kjeldahl method-not less than 0.9%N2

Raw Soy Sauce

Filter

MixingTankBottling

washing

*remove seed coats/hulls

PROCESSING PLANTPROCESSING PLANT

CRITICAL CONTROL POINTCRITICAL CONTROL POINTSTEPSTEP HAZARDHAZARD

1. Ingredient of 1. Ingredient of koji; soy bean, koji; soy bean, wheat flour.wheat flour.

moisturemoistureInsectInsectMicroorganismsMicroorganisms

2. Boiling of soy 2. Boiling of soy beanbean

Uncooked soy bean: effect growth of mould & NUncooked soy bean: effect growth of mould & N22 productionproduction

3. Innoculum3. Innoculum low Q of innoculum – effect fermentation of koji.low Q of innoculum – effect fermentation of koji.

4. Koji4. Koji uncontrolled temp:-uncontrolled temp:-

- high temp - inhibit the growth of - high temp - inhibit the growth of A. oryzaeA. oryzae

- - low templow temp. – . – growth ofgrowth of Rhizopus nigrificans Rhizopus nigrificans (black (black mould)mould)

*Best temp. in fermentation of koji *Best temp. in fermentation of koji

3030°C - 35°C.°C - 35°C. uncontrolled moisture uncontrolled moisture

- high moisture – growth of bacteria- high moisture – growth of bacteria

* Best moisture content of soy bean * Best moisture content of soy bean

43% - initial of incubation43% - initial of incubation

30% - end of incubation30% - end of incubation

CRITICAL CONTROL POINT….CRITICAL CONTROL POINT….contcont..

STEPSTEP HAZARDHAZARD

5. Concentration of brine5. Concentration of brine less 20% - enhance less 20% - enhance the growth of undesired the growth of undesired microorganismmicroorganism

6. Pasteurization6. Pasteurization incomplete – effect the incomplete – effect the flavour and the shelf life flavour and the shelf life of soy sauce.of soy sauce.

7. Cleaning and 7. Cleaning and sanitizationssanitizations

unsanitized of the unsanitized of the factory will effect the factory will effect the whole of soy sauce whole of soy sauce production.production.

CURRENT ISSUECURRENT ISSUE1.1. Before the Before the GazetteGazette of Food Regulation 1985, a lot of soy of Food Regulation 1985, a lot of soy

sauce manufacturers:sauce manufacturers:

Used saccharin as sweetener in soy sauce. Used saccharin as sweetener in soy sauce. Used recycled alcoholic bottles.Used recycled alcoholic bottles. Addition of Acid Hydrolysed Vegetable protein to accelerate Addition of Acid Hydrolysed Vegetable protein to accelerate

production of soy sauce.production of soy sauce.

Problem in meeting the requirements of the total Problem in meeting the requirements of the total nitrogen of soy sauce – because of the poor processingnitrogen of soy sauce – because of the poor processing

2.2. The main source of 3-MCPD (3-monochloropropane-1,2-The main source of 3-MCPD (3-monochloropropane-1,2-diol) in soy sauces is from the addition of acid-hydrolysed diol) in soy sauces is from the addition of acid-hydrolysed vegetable protein (acid-HVP), a savoury food ingredient vegetable protein (acid-HVP), a savoury food ingredient which has been found to contain 3-MCPD.which has been found to contain 3-MCPD.

3.3. 3-MCDP is a group of chemicals known as 3-MCDP is a group of chemicals known as chloropropanols.chloropropanols.

MAKING OF KOJIMAKING 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)

FERMENTATIONFERMENTATION

Koji and brine (16 – Koji and brine (16 – 18%) are put in 18%) are put in earthen jar under sunlight under sunlight (10 – 12 weeks).(10 – 12 weeks). Cover during the night & rainy day Moromi formedMoromi formed

SORTING Good Q of soy bean is important in making soy sauce.

During sorting – defected soy beans are separated.

BOILINGBOILING

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).

MIXINGMIXING

Cooled soy beans + Cooled soy beans + wheat flour (20%) + wheat flour (20%) + innoculum of innoculum of AspergillusAspergillus oryzaeoryzae (0.1%) in mixer, eg: (0.1%) in mixer, eg: ribbon mixer. ribbon mixer.

Aspergillus oryzae

STAGE 1 :KOJISTAGE 1 :KOJI

The mixture is spread (2cm The mixture is spread (2cm thick) onto the stainless thick) onto the stainless steel tray (with hole).steel tray (with hole).

These trays are covered These trays are covered with muslin cloth.with muslin cloth.

Incubate in koji room– Incubate in koji room– 3030°C/48°C/48 hrs/RH 90% hrs/RH 90%

At this stage – protein, At this stage – protein, oligo and polysaccharides oligo and polysaccharides are hydrolyzed – are hydrolyzed – Aspergillus Aspergillus enzymeenzyme

This results in production This results in production of fermentable sugar, of fermentable sugar, peptides and amino acids.peptides and amino acids.

Mould grow Mould grow Koji formed (greenish yellow - good Q).(greenish yellow - good Q).

Longer incubation – allow Longer incubation – allow the growth of undesirable the growth of undesirable black moulds.black moulds.

FERMENTATIONFERMENTATION Koji - put into food grade fiber Koji - put into food grade fiber

tank – equipped with cover, tank – equipped with cover, valve, pipe, filter, air flow. valve, pipe, filter, air flow.

Brine (22%) is put into this tank. Brine (22%) is put into this tank. 1:3 (koji’s wt : brine’s wt) – to kill 1:3 (koji’s wt : brine’s wt) – to kill the growth of the growth of Aspergillus oryzae. Aspergillus oryzae. moromi formed moromi formed

Lactic fermentation occursLactic fermentation occurs– Lactobacillus delbruckiiLactobacillus delbruckii & &

Pediacoccus soyaePediacoccus soyae Followed by alcoholic Followed by alcoholic

fermentation (by yeast) – fermentation (by yeast) – Zygosaccharomyces rouxiiZygosaccharomyces rouxii

Exposed to the sunlight – Bright Exposed to the sunlight – Bright and sunny weather tends to be and sunny weather tends to be more conducive in producing soy more conducive in producing soy sauce of better Q and aroma.sauce of better Q and aroma.

Exposure for 2 months – to get Exposure for 2 months – to get the min aroma of soy sauce and the min aroma of soy sauce and continuously developed. continuously developed. Recommended: ferment the Recommended: ferment the moromi at least 2 months.moromi at least 2 months.

To ensure the conc. of To ensure the conc. of the brine not less thanthe brine not less than 20% - use brinometer.20% - use brinometer.

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

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

Aroma developed Aroma developed

Moromi

BrinometerBrinometer

EXTRACTION & FILTRATIONEXTRACTION & FILTRATION

After fermentation, raw soy sauce After fermentation, raw soy sauce will be extracted and filtrated to will be extracted and filtrated to aging tank – using hydraulic/pump. aging tank – using hydraulic/pump.

In aging tank – the mixture sunned In aging tank – the mixture sunned for a week– to develop aroma and for a week– to develop aroma and flavour. flavour.

SEDIMENTATIONSSEDIMENTATIONS

The raw soy sauce will be pumped to the The raw soy sauce will be pumped to the sedimentation tank. sedimentation tank.

Sampling for the total nitrogen using Sampling for the total nitrogen using Kjeldahl method – the total nitrogen Kjeldahl method – the total nitrogen supposed to at least 0.9% before mixing supposed to at least 0.9% before mixing with other ingredients and pasteurization with other ingredients and pasteurization – may lost during heating.– may lost during heating.

The processed soy sauce must contains at The processed soy sauce must contains at least 0.6% total nitrogen (in Food least 0.6% total nitrogen (in Food Regulation 1985).Regulation 1985).

MIXING MIXING Raw soy sauce with Raw soy sauce with

sufficient nitrogen will be sufficient nitrogen will be pumped to the mixing tank.pumped to the mixing tank.

Brine (18%), sugar, caramel Brine (18%), sugar, caramel and food additive will be and food additive will be added in the ratio 1:2 (vol added in the ratio 1:2 (vol ingredient : vol raw soy ingredient : vol raw soy sauce).sauce).

Caramel and sugar Caramel and sugar to to condense and sweeten the condense and sweeten the sauce.sauce.

Caramel Caramel calouring agentcalouring agent Food preservative ; Food preservative ;

eg : benzoic acid – not more eg : benzoic acid – not more than 1000 ppm (Food than 1000 ppm (Food Regulation 1985).Regulation 1985).

Flavour enhancer also can Flavour enhancer also can be added (eg: MSG)be added (eg: MSG)

PASTEURIZATIONPASTEURIZATION

Raw soy sauce will be Raw soy sauce will be pasteurized at 80pasteurized at 80°C/30 °C/30 min in mixing tank min in mixing tank equipped with equipped with

stirrer and jacketed stirrer and jacketed thermal oil (to thermal oil (to prevent – burnt prevent – burnt and crusty), orand crusty), or

Plate heat Plate heat exchangerexchanger..

BOTTLINGBOTTLING The processed soy The processed soy

sauce will be filled sauce will be filled into sterilized into sterilized bottles using high bottles using high pressure vacuum.pressure vacuum.

Soy sauce – Soy sauce – capped, capped, labelledlabelled and distributed.and distributed.

Conclusion Conclusion

All of the food potential to be All of the food potential to be fermented (naturally/ modified), but fermented (naturally/ modified), but if there’s no controlled handling, if there’s no controlled handling, proteolitic and lipolytic will replace proteolitic and lipolytic will replace the process and spoiled the food.the process and spoiled the food.

TODAY’S THORN TODAY’S THORN

IS TOMORROW’S FLOWERIS TOMORROW’S FLOWER

CHARACTERISTICS OF FIBRE CHARACTERISTICS OF FIBRE TANKTANK

Food gradeFood grade recyclable and allows for easy recyclable and allows for easy

disposal disposal high-temperature performance high-temperature performance corrosion resistance corrosion resistance