Kecap Ikan_AnantaLevinaSavitri_13.70.0003_C3_UNIKA SOEGIJAPRANATA
Indonesian Soy Sauce (Kecap)
Transcript of Indonesian Soy Sauce (Kecap)
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INDONESIAN SOY SAUCE (KECAP)FERMENTATION
SardjonoDepartment of Food and Agricultural Product Processing TechnologyFaculty of Agricultural TechnologyGadjah Mada UniversityYOGYAKARTA, INDONESIA
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HEALTH BENEFIT OF JAPANESE FERMENTED SOYSAUCE
http:// www. Raysahelian.com/soysauce.html
Promote digestion
Antimicrobial activity againts : Staphylococcus aureus, Shigellaflexneri, Vibrio cholera, Salmonella enteridis, Esherichiacoli
Antihypertensive effects
Inhibit benzopyrene induce neoplasia
Antioxidant activities, anticarcynogenic effects
Inhibit histidine decarboxylase anti alergy
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KECAP IS LIQUID SEASONING, BROWN
BLACK IN COLOR, PRODUCED FROM
FERMENTED SOYBEAN, WITH ADDITION OF
COCONUT SUGAR
PRODUCED BY TWO STEP OF FERMENTATION :
1. MOLD FERMENTATION ( solid substrate fermentation,with different of mold used ) produced KOJI
DEGRADATION OF POTEIN
2. BRINE FERMENTATION , (22% NaCl solution ,natural
selective microorganisms: lactic acid bacteria and
halophyilic yeast )
produced MOROMIPROTEIN DEGRADATION AND EXTRACTION, FLAVOR
DEVELOPMENT
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KOJI FERMENTATION
Degradation by extracellularmold enzymes
Extraction : enzymatic, physical?Flavor development :
microbiological? Chemical?
FERMENTATION TIME ??
MOROMI FERMENTATION
KEY OF PROCESS FOR
SOY SAUCE FACTORY
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SIMPLYFIED PROCESS FLOW SHEET OF KECAP
PRODUCTION
soybean
COOKINGMOLD FERMENTATION
DRYING THE KOJI
BRINE FERMENTATION
SEPARATION OF BEANCOOKING / BOILING
starter or wildfermentation
Coconut sugar
NaCl solutIon (22%)
kecap
soybean
koji
moromi
OPTIONAL
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KOJI MOROMI
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BRINE FERMENTATION (MOROMI)BRINE FERMENTATION (MOROMI)
Salt concentration, pH
and temperature
enzymes activity,population ofmicroorganism
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KoikuchiKoikuchi--shoyushoyu
(representative fermented soy sauce in Japan)(representative fermented soy sauce in Japan)
Soybean orDefatted soybean roasted wheat (50%)
cooked
KOJI FERMENTATION starter
lactic acid bacteria MOROMIyeast FERMENTATION
(pH, temp)
PRESSED cake
PASTEURIZED
koikuchi-shoyu
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KOJI FERMENTATION(traditional method)
Small factory Medium / big factory2 4 DAYS FERMENTATION
Un-control condition
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MYCOFLORA AND AFLATOXIN IN KOJI FROM SEVERALTRADITIONAL FACTORIES (Sardjono, 1985)
Spesies jamur 1 2 3 4 5 6 7 8 9 10
Aspergillus
A. flavus/parasiticus 34 20 50 50 30 50 22 16 18 16
A. niger var. niger 0 0 0 0 0 0 0 0 0 0A. versicolor 0 14 2 0 0 0 0 0 0 0
A. alliaceus 32 0 0 0 0 0 0 0 0 0
A. fumigatus 0 0 0 0 50 0 0 28 0 0
A. sojae 0 32 0 32 30 58 0 38 36 30
A. oryzae 0 0 58 0 0 0 100 0 38 30
Eurotium
E. rubrum 0 30 0 30 40 38 58 54 62 50E. chevalieri 60 32 30 42 32 20 0 46 42 54
E. amstelodami 0 46 40 32 32 0 46 0 0 0
Fusarium
F. moniliforme 38 48 0 32 36 0 0 0 0 0
F. chlamydosporum 0 0 36 0 32 0 0 0 0 0
F. poae 42 0 0 36 0 0 0 0 0 0
PenicilliumP. echinulatum 1 4 0 0 0 0 0 0 0 0
P. melinii 0 0 0 2 0 0 0 0 0 0
Genera lainnya
Rhizopus oryzae 20 48 30 48 48 70 20 68 62 100
Curvularia lunata 0 0 12 0 0 0 0 0 0 0
AFB1 (ppb) 14 15 ND 36 16 13 ND 30 ND ND
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Aspergillus sojae
Aspergillus oryzae
COMMON MOLD USED IN SOY SAUCE FERMENTATION
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Aspergillus flavus
Aspergillus oryzae
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Aspergillus oryzae
Aspergillus sojae
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konidia
fialida
metula
vesiklua
BISERIATE UNISERIATE
GeneraAspergillus
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Aspergillus flavus
Medium Skim Milk Agar Medium AFPA
Aflatoxigenic test
Clear zone
Orange inreverse color
Proteolytic test
Simple method for selection of mold strain
Pepton 10.0 g/l
Yeast extract 20,0Ferri Amm. Sitrat 0,5
Dichloran 0,002
Agar 15,0
pH : 6,2 + 0,2
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IMPORTANT FACTORS IN KOJI FERMENTATIONIMPORTANT FACTORS IN KOJI FERMENTATION
1. Softness of beans promotes the fungal growth andpenetration of mold important to protein digestibility and formolnitrogen production
2. Molds used for koji productionAspergillus oryzae : produce amylase and protease enzymesAspergillus sojae : produce more complex of protease enzymes
and produce glutaminase3. Fermentation conditions :Relative Humidity (affect water activity of bean)
important to fungal mycelia growth andprevention on spore formationOxygen important to fungal growth and affecting thespore formationTemperature important for production and activities of enzymes
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The role ofThe role of AspergillusAspergillusoryzaeoryzae andand AspergillusAspergillussojaesojae
Aspergillus oryzae : amylase and protease enzymes
sugars important to Lactic acid bacteriaand yeast in brine fermentationAspergillus sojae : more complex of protease enzyme andproduce glutaminaseBeans contained a lot of glutamine and glutamic acid as amino acid
residue
easily change glutaminaseglutamic acid
pyroglutamic acid meaningless as flavorcomponent
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ENZYMES PRODUCED DURING KOJI FERMENTATIONENZYMES PRODUCED DURING KOJI FERMENTATION
ProteinaseAlkaline (the highest proportion and activity) Optimum pH : 10.5Semi alkaline 8.3Neutral I 7.0Neutral II 6.0
Acid I 3.2Acid II 3.0
Acid III 3.0
Acid carboxypeptidasesAcid carboxypeptidase (I, II, III and IV ) 3 4
Leucine aminopeptidase(Enzyme I, II, III, IV, V, VI, VII and Arylamidase) 5.0 8.5
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CEREAL PROTEIN
PEPTIDES
AMINO ACIDS
GLUTAMINE PYROGLUTAMIC ACID
GLUTAMIC ACID
ALKALINE PROTEINASE
SEMI ALKALINE PROTEINASE
NEUTRAL PROTEINASE I AND IIACID PROTEINASE I-III
ACIDCARBOXYPEPTIDASES I IV
LEUCINE AMINOPEPTIDASES I - VII
GLUTAMINASE((Aspergillus sojae)
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THE DIFFERENCES BETWEEN MOROMI FERMETATION OFTHE DIFFERENCES BETWEEN MOROMI FERMETATION OFJAPANESE SHOYU AND KECAPJAPANESE SHOYU AND KECAP
JAPANESE SHOYU KECAPMoromi are put inside Moromi are put outside
( no sun light ) ( direct sun light )
Low temp. in the beginning higher temperaturefermentation
slower Lactic bact growth faster Lactic bactgrowth
slower pH change faster pH change
higher protease activity lower protease activity
higher formol nitrogen lower formol nitrogen
No pellicle yeast pellicle yeast( because Lack of sugars)
not necessary sun light sun light inhibit their growth
Pellicle forming yeast cause off-flavor in Japanese shoyu
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Advantage and disadvantaged drying ofAdvantage and disadvantaged drying of kojikoji
DRYING and CRUMBLING OF KOJIDRYING and CRUMBLING OF KOJI
Removed someRemoved some DecreasedDecreased glutaminaseglutaminase Protease andProtease and LeucineLeucineVolatile compoundVolatile compound activityactivity amino peptidaseamino peptidasediminished stronglydiminished strongly
diminished production ofdiminished production of
glutamicglutamic acid ammonia,acid ammonia, formolformol nitrogennitrogen
lower buffering capacitylower buffering capacity
Inhibitory effectsInhibitory effectsonon microbmicrob. growth. growthIn brine fermentation?In brine fermentation? Fermentation not moreFermentation not more allowedallowed
affecting the production ofaffecting the production ofmoromimoromiflavorflavor
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Whole soybean : black/yellow?
RAW MATERIAL :
Defatted soybean
DIFFERENT CHARACTERISTIC OF FINAL PRODUCT?
Different in : 1. Composition oil in final product?Amino acid in final product?
2. Physical structure solid substratefermentation?
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R3
R1
R2
H OH
OCH3
CH=CHCOOH
R1 R2 R3
H H -O-CH(COOH)-CH(OH)COOH
OH H -O-CH(COOH)-CH(OH)COOH
OH OH -O-CH(COOH)-CH(OH)COOH
H H H(Daidzein)
(Isoflavon derivatives)
(Isoflavon derivatives)
(Isoflavon derivatives)
Ferulic acid
DEFFERENTIATION OF NON-VOLATILE COMPOUND OF SOY SAUCEFROM WHOLE AND DEFATTED SOYBEAN
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KOJI FERMENTATIONKOJI FERMENTATION
Solid Substrate FermentationSolid Substrate Fermentation
Principles?Principles?
Design of Bioreactor?
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Environmental change during Solid SubstrateEnvironmental change during Solid SubstrateFermentation and their effectsFermentation and their effects
waterMold growthProduct formationSpore formationMycotoxin/pathogen
heat
Mold growth
Product formationEnzyme activitiesMycotoxin
pH Enzyme activitiespathogen
O2, CO2
Mold growthProduct formationMycotoxin
Biomass determination:Viable count methodMetabolic activities
OCR and CPRlost of solidproduct formation
Biomass componentN and proteinDNAGlucosamineErgosterol
Other phenomena
Growth kinetics and the model of growth in SSF?
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AERATIONHEAT
REMOVAL
HUMIDIFY( aw)
agitation
OCR and CPRTransfer of O2 into interparticlesDiffusion of O2 substrate particlesTransfer of CO2 into interparticles
Remove by conduction
Remove by convectionJacket system (externally?)Chilled water
Chilled water?
Fungi are susceptible for shear process
Mixing frequency depend on void fraction
Handling of
substrate
pH?
PROCESS OPERATION IN
Solid Substrate Fermentation REACTOR
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A. oryzae, soybean
0
0.001
0.002
0.003
0.004
0.005
1 2 3 4 5 6 7 8
Fermentation time (days)
OCR,C
PR
(u
mol/s/mg)
0
24
6
8
10
12
Gluc
osamine(mg
)
OCR(umol/s/mg) CPR(umol/s/mg) Gluc(mg)
Sardjono, et al., 1998
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A. oryzae, soybean
0
2
4
6
8
10
12
14
1 2 3 4 5 6 7 8Fermentation time (day)
OCR
,CPR(mmol/
mginitialdrymatter)
0
0.02
0.04
0.06
0.08
0.1
0.12
0.140.16
0.18
Drym
atterloss
(g/ginitialdrymatter)
OCR(mmol/mg)CPR(mmol/mg)Dry matter loss(g/g initial dry matter)
Sardjono, et al., 1998
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Glucosamine, wateractivity and sporeconcentration
Sardjono, et al., 1998
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GlucosamineGlucosamine and enzymes activitiesand enzymes activities
Sardjono, et al., 1998
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KOJI FERMENTATION(traditional method)
Small factory Medium / big factory
2 4 DAYS FERMENTATION
Un-control condition
Sardjono, et al., 1998
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THE SURFACE OF DRIED KOJITHE SURFACE OF DRIED KOJI(After crumbling the(After crumbling the kojikoji to remove spore and mycelia)to remove spore and mycelia)
FRACTURED / CRACKED
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Batch type with rectangular perforated plateBatch type with rectangular perforated plate
Solid Substrate Fermentation equipmentSolid Substrate Fermentation equipment
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Materials turn over?
Heat, CO2 removal only
on the surface of substrate
Limited aeration
Batch type with rectangular perforated plateBatch type with rectangular perforated plate
Solid Substrate Fermentation equipmentSolid Substrate Fermentation equipment
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BRINE FERMENTATION (MOROMI)BRINE FERMENTATION (MOROMI)
NaCl concentration, pHand temperature
enzymes activity,population ofmicroorganism 5 6 MONTH FERMENTATION
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MOROMI FERMENTATIONMOROMI FERMENTATION
Continuing the activity of koji enzymes:
pH of moromi , temperatureExtraction of protein from koji:
salt concentration, time of fermentation
Flavor development / formation:
pH, temperature microbial activityBacillus died ( 2 months) ???
Lactic acid bacteria at first, followed by salt tolerantyeast
( pH 4 - 5 )Detoxification of mycotoxins, if any
cyclopiazonic acid ( 3 months), AFB1?
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SALT TOLERANT LACTIC ACID BACTERIA ANDSALT TOLERANT LACTIC ACID BACTERIA AND
OSMOPHYLIC YEASTOSMOPHYLIC YEAST
Lactobacillussp
Saccharomyces ruoxii
MOROMI
Pediococcussp
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What must be happened during moromi fermentation?
Prepare the condition for the activity of koji enzymes
The growth of microorganisms which play an important roleduring brine fermentation
Extraction of soluble protein from the fermented bean (koji)
To prevent the pH from declining too quickly as a result of
sudden growth of Lactic acid bacteria
A rapid drop in pH obstructs the action of proteolyitic enzymesand glutaminase enzyme of koji decreasing the soluble
protein and glutamic acid in final products
pH and salt concentration the growth of Lactic acidbacteria and yeast which are important during brine
fermentation
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Aspartic Acid
Glutamic Acid
Serin
HistidinGlysin
Arginin
Alanin
Tirosin
Metionin
Valin
FenilalaninIsoleusin
leusin
Lysin
43,722
79,110
68,703
22,63927,294
32,038
10,006
26,45
25,08452
24,06604
38,2510842,85296
32,61771
63,9268
AMINO ACIDS ppm
Umami taste
Mengalamitransaminase
oleh bakteri
asam laktatmenjadikomponenflavor moromi
AMINOA
CID
S
IN
MOROMI
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COMPOUNDS
Propylene glycol
1-Propanol, 2-methyl-
2-Pentanol
1-Butanol, 3-methyl-
1-Butanol
2-Octanol
1-Octen-3-ol
Acetic acid
Propanoic acid
Propanoic acid, 2-methyl
Butanoic acid
Butanoic acid, 3-methyl-
Butanoic acid, 2-methyl-
Phenol, 2-methoxy-
Phenol
2(3H)-Furanone, dihydro-
2-furanmethanol
Furfural
Benzaldehyde
pyrones
Maltol
unknown
unknown
unknown
unknown
unknown
unknown
unknown
unknown
furans
aldehyde
unknown compounds
alcohols
acids
phenols
furanones
odorant
flowery
sweet, caramel like
condiment-like
sour
sweet, caramel-like
sour, spoiled like
sweet sour
green-like
stale (apek)
astringent, stale (ape
cereal like
mushroom-like
spoiled-like
soysauce like
rancid
roasted potato
nutty
condiment like
cheesy
KO
MPONEN
FL
AVOR
MORO
MI
YANGB
ERHASILDIDETEK
SI
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3-methyl -1-butanol and 1- propanol-3-methyl, might attribute to a powerfullmeaty and soy sauce odor and flavor at high dilution.
Furan methanol, a powerfull aromatic compound at very low threshold value.
2-methyl propionic acid also produce stronger flavor (Zhang Yanfang and TaoWenji, 2009).
Rich in amino acid able to get more flavor compound through transamination bytransaminase enzyme of Lactic Acid bacteria (Mengjin Liu et al., 2008).
SENYAWA YANG MEMBERIKAN KONTRIBUSI PADA
FLAVOR PENTING PADA MOROMI
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?