1. Microbial Production of Ganjang and Dyeonjang
(Soy Sauce)
Some photos from Google
Some videos from Youtube

2. 1. Classification of Ganjang
Definition of Ganjang soy sauce in Korean codex (식품공전)
1. Fermented sauce mixed with NaCl solution and Meju (메주) or koji solution
containing proteins and carbohydrates
2. Sauces processed with filtrated materials of hydrolyzed soybean solution by
enzymes and acids
Classification 1
Category
Source
Microorganism
Soy sauce
Korean Ganjang
Soybean
Bacillus subtilis
재래간장 (막간장, 겹장)
Japanese Ganjang
Soybean, starch
Aspergillus oryzae
양조간장, 다마리장유, 혼합장유
Fish(魚) Ganjang
fish
protease
어간장(한국), Patis(필리핀), Budu(말레이시아), Nuoc-mam(베트남)

3. 1. Classification of Ganjang
1. Fermented soy sauce (양조간장) : soy sauce from fermented solution, which comes from a mixture of NaCl and koji solution made with plant protein (soy bean) only or a mixture of starchy sources (rice, barely and wheat)
2. Mixed soy sauce (혼합간장) : a soy sauce mixture of original fermented solution and acid-hydrolyzed solution made of soy bean
(1) Current type of fermented soy sauce
: a soy sauce extracted from fermented solution consisting of original solution of acid-hydrolyzed soybean, and plant proteins or starch sources
(2) Mixed soy sauce without additional fermentation steps
*Quality standard of 혼합간장 : dependant on the content of fermented soy sauce
(>60% 특급, >40%  고급, >20%  표준)

4. 3. Healthy effects of dyeonjang/ganjang
Components of soybean
1. Protein - 30~50% in total mass
- amino acid supplement for Asian people
2. Fat ~20% in total mass
- unsaturated fatty acids of linoleic acid and linolenic acid
 prevention of cholesterol attachment on veins
and removal of cholesterol in arteries
3. Carbohydrate – 25~35% in total mass
- no starch. Containing rare sugars such as raffinose and stachyose
 Growth promotion of intestinal lactic acid bacteria
Suppressing caries production
 Reduction of blood cholesterol level. Prevention of colon cancer
4. Inorganic matters – 4~6% in total
- 80% is potassium and phosphorus  alkali foods (=foods containing a large amount of inorganic matters) (tofu, milk etc.)
5. Others – vit B1 and B2 are sufficient, no vit C.
- abundant Vit. E  tocopherol : anti-aging effect

5. 3. Healthy effects of dyeonjang
Excellent composition of amino acid in dyeonjang
1. Soybean contains 16 amino acids among total 20 amino acids.
- It has 8 essential amino acids such as threonine, valine, leucine, isoleucine,
phenylalanine, lysine, methionine, tryptophan
- Lysine & leucine are insufficient in rice and barely
2. Composition change by fermentation
 Making new amino acids and
changing the composition by
microbial fermentation

6. 3. Healthy effects of dyeonjang
Effects of methionine on caring fatty liver
1. Normal liver : destruction of fat and provision of fatty acid to other organelles
Abnormal liver : unable to decompose fat which accumulates inside the liver
(reason : nutrient imbalance, excess alcohol
2. Methionine is essential for fatty acid metabolism.
3. Intake of methionine  choline synthesis for fatty acid hydrolysis
 activation of fatty acid hydrolysis  removal of fat on fatty liver
4. Additional effect : removal of aldehyde from alcohol and nicotine toxin in tobacco
Reduction of cholesterol level by soybean lipid
1. Lecithin(레시틴) : one of phospholipid, important in cell structure and metabolism
- Present in outer membrane of nerve fiber
 When its level is decreased, memory ability will be reduced.
2. Lecitin and linoleic acid : prevention of cholesterol attachment on blood vein
 prevention of the arteries hardening and high blood pressure

7. 3. Healthy effects of dyeonjang
Anti-cancer effect of dyeonjang
1. Intake of dyeonjang reduced the occurrence of stomach cancer by 30% (Japan).
2. Traditional jang sauces prevented gene mutagenesis causing cancers (Korea)
- Dyeonjang (70%), Kochujang (50%), Ganjang (30%)
3. Dyeonjang has the cancer prevention effect and growth inhibition of cancer cells.
- Cancer prevention effect
: traditional Korean dyeonjang >> Japanese miso dyeonjang
- All toxins produced by fungi are destroyed in fermentation, washing,
submerging and maturing processes.
Other effects
1. Treatment of high blood pressure – both dyeonjang and chunggukjang
2. Acceleration of intestinal lactic acid bacteria growth : prevention of toxin-producing microorganisms and acceleration of lactic acid bacteria growth such as Bifidobacteria and Lactobacillus

8. 4. Korean Soy Sauce (1) Production Process
1. Making meju + Fermentation & Maturation
2. Important in controlling the content of NaCl
Soybean
Washing+Submerging
Raw
Soy sauce
Boiling
Soy sauce
Steam cooking
Submerging
Maturation
Separation
Meju mass
Maturation
Dyeonjang
Grinding+Shaping
NaCl
Solution
NaCl
Hanging
NaCl
Meju

9. 4. Korean Soy Sauce (1) Production Process
Pretreatment of soybean
- Washing and submerging in water for 8 ~12 hrs
- Boiling soybean in hot water for more than 2 hrs  Protein denaturation
Making Meju
- Grinding with 절구 or 초파기 (직경 8~10 mm)
- Shaping : It is regionally different (전라도 메주: 15*15*20cm)
- Drying its surface on the rice straw for about 2 weeks
- Hanging : tiding Meju with rice straw and hanging on the ceiling in winter
 Microorganisms are inoculated naturally and grown on the surface of and inside Meju.

10. 4. Korean Soy Sauce (1) Production Process
Making Meju
- Microorganisms working for Meju fermentation
Class
Distribution
Microorganism
Characteristics
Fungi 1%
Rizopus sp.
Mucor abundans
Scopulariopsis brevicaulis
Aspergillus oryzae
Penicillium lanosum
Aspergillus sojae
Mainly present in the surface of Meju
Hyphae grows on the cracking fissure of Meju.
Bacteria
99%
Bacillus subtilis
Bacillus pumilis
Mucor griseocyanus
Growth on the surface of and inside Meju
Only bacteria can grow inside Meju.
Secretion of strong proteolytic and carbohydrate hydrolysis enzymes
Yeast
0.01%
Rhodotorula flava
Torulopsis dattila
Involved in flavors and tastes

11. 4. Korean Soy Sauce (1) Production Process
Submerging & Maturation
- Submerging the Meju in NaCl solution
- Generally late winter and early spring
- Formulation of NaCl solution : Strongly related with microbial growth and maturation of sauce
Traditional method : water 4 두(斗) + NaCl 1 두(斗) similar to the current method (Be’19o)
- Method :
Washing the surface of Meju & dividing to 2~3 pieces & fully dry by sun
After submerging the Meju, spread NaCl powder on the Meju which is over the surface of the NaCl solution  Prevention of spoiling M/O
During fermentation and maturation, open the cover in the sunshine
 Prevention of lowing flavor and taste

12. 4. Korean Soy Sauce (1) Production Process
- Microorganisms related to maturation
Aerobic bacteria : Bacillus species
Salt-resistant lactic acid bacteria : Pediococcus halophilus, L. casei, L. plantarum, Le. mesentoroides
Salt-resistant yeast : S. rouxii, S. acidofaciens, Torulopsis dattila
Lactic acid bacteria and yeast are related to unique flavor and taste of soy sauce.
Change of microbial population
during maturation of Korean soy sauce

13. 4. Korean Soy Sauce (1) Production Process
Filtratio & boiling (pasteurization)
- Filtration : Separation of solution part (soy sauce) and solid part (meju)
- Raw soy sauce : Soy sauce before boiling. Contains enzymes and M/O.
Young taste and flavor
- Boiling : boiling raw soy sauce for 10 ~20 min
Improvement of storage time, flavor and taste
Concentration of soy sauce
Color of soy sauce : Melanin and melanoidin from amino acid residues
* For acid-hydrolysis soy sauce, caramel (like 물엿) is used for coloring.
Flavor of soy sauce : alcohols, ketones, aldehydes, volatile acids, esters, phenols
Traditional taste of soy sauce: β-mercaptopropyl alcohol

14. 4. Korean soy sauce (2) composition
Various kinds of Korean soy sauce
Major component : amino acids, flavors, alcohols, organic acids, salt
Amino acid : Its content and composition varies according to making process
Salt content : 19~25%
Total nitrogen : 0.36~0.95%
Organic acid : butyric acid propionic acid, acetic acid, formic acid
(total acid: 0.5%)
Flavor & taste : Aromatic and sweet taste, salty taste
General composition of sauce made of soybean meju (scale, %)
Water
Ash
Protein
Lipid
Fiber
Carbohydrate
Salt
Soybean
9.0
4.6
38.0
20.7
4.7
23.0
0.2
Traditional meju
3.2
4.1
43.0
17.8
5.8
6.1
0.3
Korean soy sauce
68.9
24.2
4.4
0.1 -
2.3
23.6

15. 5. Japanese soy sauce Japanese soy sauce (Japanese Shoyu)
: Major sources are soybean and wheat.
Making Japanese soy sauce
: Koji making (Aerobic growth of fungi to produce hydrolyzing enzymes)
+ Fermentation & Maturation (lactic acid bacteria + yeast)
: Koji  enzyme sources for hydrolysis of protein and carbohydrates in soybean
M/O : Aspergillus oryzae or A. sojae
Types of Japanese soy sauce
Koikuchi type : more than 90% of Japanese soy sauce. Dark color
- Source : mixture of the same amount of soybean and wheat
- Hydrolysis : A. oryzae
- Fermentation & maturation : lactic acid and alcohol fermentation
- Sterilization : High temperature condition
Usukuchi type : less than 10%. 1.2% total nitrogen content. Bright color

16. 5. Japanese soy sauce (1) Material
Soybean : supply of protein and fat, essential fatty acid, lecitin, flavonoid
- More than 92% is imported : Soybean oil (75%) + food product (25%)
Skimmed soybean : Used after oil extraction with n-hexane
Wheat grain : starch source (more than 80%)
- Unique flavor and taste are formed
Salt & water : NaCl (more than 90%) + various salts (CaSO4, MgSO4, MgCl2, KCl)
- MgCl2 : bitter taste
- Use NaCl salt with a low content of iron

17. 5. Japanese soy sauce (2) Process
Source treatment + fermentation + shaping
Source : soybean+wheat grain
Fermentation starter : koji
Additional inoculation
: P. soyae, S. rouxii
Filtration & boiling
Process for making koikuchi-tyype shoyu

18. 5. Japanese soy sauce (2) Process
Source treatment : washing, submerging, cooking
Cooking soybean
- process for easy hydrolysis of soybean protein by proteases from koji fungi
- Protein denaturation by cooking increases the proteolytic ability of protease
- N.K.(Nippon Kikkoman)-type cooking (평압증자법, 회전증자+진공냉각)
Koji making
After processing wheat grain (cooking), Aspergillus species is inoculated and grown in processed wheat grain (and cooking soybean) for production of protease and amylase
Aspergillus strains (국균) : A. oryzae, A. sojae

19. 5. Japanese soy sauce (2) Process
Submerging
Koji is added into a container with salt solution
Ratio : kogi 1L + 1.1~1.3L salt solution
Use of salt solution at -5oC : prevention of acid-forming M/O
 remaining of hydrolysis enzymes
Content of salt : NaCl 23.2~24.6%
Salt content in final submerging solution : 16.5~18%
Maturation : fermentation by salt-resistant M/O
P. sojae, L. delbruckii, S. rouxii
Mixing and Heating
Mixing : control of total nitrogen content by mixing with sub-materials (sugarcane, caramel) or acid-hydrolysis soy sauce
Heating : destroy of residual enzyme, sterilization, harmony of taste and flavor, 60~70oC, 10 min

20. 6. Acid-hydrolyzed Soy Sauce
Acid-hydrolyzed soy sauce : amino acid soy sauce or chemical soy sauce
Sauce solution : Amino acid solution by hydrolysis of soybean and wheat protein with hydrochloric acid, neutralization with alkali and concentration
18%
60~70시간 분해
분해취 억제
NaOH, pH 4.8~5.2
산분해간장 제조공정

21. 6. Acid-hydrolyzed Soy Sauce
Components :
- Amino acid : No tryptophan. Low Serine/threonine. High arginine
- Organic acid : High level of oxalic acid, formic acid, levulinic acid (chemically)
Low level of lactic acid, succinic acid, propionic acid (biologically)
<양조간장과 산분해간장에서 총질소 및 유기산 함량 (단위, mg%)>

22. 7. Specification of soy sauce