1.

0 INTRODUCTION Soy sauce is a condiment in which it is gone through the same method with miso, which is fermented and aged product. The soy sauce is fabricated by fermenting soybeans with Aspergillus oryzae or Aspergillus sojae moulds along with water and salt. The soy bean which is salted and fermented food are created back in the days in order to preserve for longer shelf life and flavour. The soy sauce is important especially in for South East Asian cooking which comprise mostly salted and fermented food product too, where it is used in cooking and as a condiment. It originated in China 2,800 years ago and spread throughout Asia. In more recent times, it is also used in Western cuisine and prepared foods. All varieties of soy sauce are salty, earthy, brownish liquids intended to season food while cooking or at the table. The sauce is in dark brown liquid, in which it is table at ambient temperature and does not need refrigeration due to its lower water activity and high salt content. Generally soy sauce is made from a mixture of soy beans, wheat, salt and yeast. Some types of soy sauce also include fermented rice (Kome Koji) or Amazake to give sweetness to the flavour. Mass produced soy sauce is made from defatted soy residue (soy from which the oil has been extracted), which makes the soy sauce ferment a lot faster than using the whole bean. After the fermentation, which yields fermented soybean paste, the paste is pressed, and two substances are obtained: a liquid, which are the soy sauce and a cake of (wheat and) soy residue, the latter being usually reused as animal feed. Most commonly, a grain is used together with the soybeans in the fermentation process, but not always. Also, some varieties use roasted grain.

1.1 TYPE OF SOY SAUCE 1

There are various types and brands of soy sauce which available in market due to the extensive use in flavouring in East Asian and South East Asian cuisines. Basically, there are two types of soy sauce which normally used in cooking which are light and dark, or sweet and salty. Dark soy sauce is aged longer than light soy sauce, giving it a brownish black colour and much thicker texture. Light sauce meanwhile has a lighter colour and saltier in flavour. Sweet and salty soy sauce as its name suggest, providing sweetish or salty flavour for cooks. Soy sauce generally poses the same appearance, but depending on the regions and origin, it may have different in taste, consistency, fragrance and saltiness. Below are the type and characteristic of soy sauce based on the region origin. China

Figure 1: Chinese soy sauce Majority of Chinese soy sauce was made from soybean and seldom use other grains. It can be divided into two classes based on the manufacturing process in which it is brewed or blended. Brewed soy sauce was made brewed directly from a fermentation process using wheat, soybeans, salt and water without additional additives. The result from the brewing normally will produce light soy sauce. Blended meanwhile are finished brewed soy sauce in which later will be mix with additives and produce dark and thicker soy sauce.

Japan 2

Figure 2: Japanese soy sauce Japanese soy sauce can be divided into five main category based on their differences of ingredients and production method. Japanese soy sauce does not include wheat as their main ingredient where impact on the sweeter taste than Chinese soy sauce. Certain soy sauce may be added with small amount of alcohol as natural preservative, white wine, red flavour or beef stock, fish stock in which produce different taste of soy sauce. Categories of Japanese soy sauce are Koikuchi which is thick in flavour, Usukuchi that has weaker taste, Tamari that have richer flavour and darker colour, Shiro is white soy sauce and Saishikomi that is double brewed and have darkest and strongest taste.

Korea 3

Figure 3: Korean soy sauce Korean soy sauce is produced from the Korean fermented soybean paste that have thin and dark brown in colour which have different degree of saltiness based on the producer. Indonesia

Figure 4: Indonesian soy sauce Soy sauce or known as kecap have three varieties which is Kecap Asin (salty soy sauce), Kecap Manis (sweet soy sauce) and Kecap Manis Sedang (medium sweet soy sauce). Salty soy sauce is quite similar to Chinese light soy sauce but usually thicker and stronger in flavour. Sweet soy sauce is thicker and sweeter due to addition of palm sugar and medium sweet soy sauce is categorized in between of both soy sauce which has less thick consistency and saltier.

Philippines

Figure 5: Filipinos soy sauce The Philippine soy sauce is similar to Japanese soy sauce which have thinner texture and saltier than most of soy sauce product in South Asian countries.

Malaysia

Figure 6: Malaysian soy sauce Malaysian use the same dialect with Indonesian by calling soy sauce as kicap, and produced in two types which is kicap lemak (rich fat soy sauce) that similar to sweet soy sauce and kicap cair (thin soy sauce) equivalent to salty soy sauce. Taiwan

Figure 7: Taiwanese soy sauce Taiwanese soy sauce use the black bean and known as black bean soy sauce and took around 6 months to produce. 1.2 SOY SAUCE GRADE Soy sauce in other regions is normally graded based on their texture or taste but in Japan, soy sauce is carefully graded based on their quality and production process. Japanese highest grade of soy sauce called as Special grade where it is exclusively made by the fermentation process and marketed under the Japanese Agricultural Standard (JAS) for the grade. This grade is characterized by a pleasant aroma, complex flavour and deep reddish brown colour. The second grade is Upper grade soy sauce where they have lighter reddish brown colour and milder aroma and flavour. The third grade is Standard grade which is characterized by slightly higher amino acid content and have lack of aroma. Special and Upper grades produced from equal amount of wheat and soy beans meanwhile for Standard rely on soy bean and use small amount of wheat.

2.0 SOY SAUCE PROCESS 6

There are two type of production method to produce soy sauce which is brewing and hydrolysis process. Brewing method use the combination of soy beans, wheat grain, water and salt. It consists of four (4) steps which are koji making, brine fermentation, refinement and pressing, lastly pasteurization. The second production method is the hydrolysis where the defatted soya bean is combined with food grade hydrochloric acid in a pressure cooker. The process goes through neutralization, refinement, treatment and filtration. The hydrolyzed soy sauce is lack in aroma and flavour and normally used to make less expensive and lower quality of soy sauce.

2.1 STEP IN MAKING SOY SAUCE

2.1.1 BREWED METHOD Brewing method is very famous among the coffee maker or even the coffee drinker because by brewing the coffee, it can give the best aroma and taste for the coffee. Same like coffee, the soy sauce also use brewing method to get the original taste and to get the best aroma of the soy sauce. The method will help in preserving the colour, taste and aroma in which producing the high quality soy sauce. Brewing is the traditional method of making soy sauce and in this method; it consists of four steps which are koji making, brine fermentation, refinement and pressing and also pasteurization.

1. Koji making

Figure 8: Koji Making In the first step of brewing method, the soybean and wheat grains are added with koji which is the source of proteolytic enzymes for converting soybean proteins into peptides and amino acids, and amylase for hydrolyzing gelatinized starch into simple sugars. The substances converted by the enzymes in Koji become the nutrients for yeasts and lactic bacteria in the subsequent brine fermentation. During this step, the soybean and the wheat grains have been carefully selected and have been washed. Than the soybeans that have been washed are soaked in water to increase the moisture content and will be steam for about 180 C until boiling for 4 hours. Upon this process, the soaked soybeans containing 30% to 45% moisture were cooked with saturated 6-7kg cm 2gauge pressure cooking. The wheat grains contains 8% moisture is heated with continuous roasting on the pan roast for 25 minutes at 150 C at 20-30 second in atmospheric pressure and the wheat grains that have 8

been roasted will be crushed into several pieces in order to let yeast to penetrate the wheat grains during fermentation process. When the steam soybeans are ready, it will be speared into a large tray to let the soybean cold until 33C. Then the cold soybean will be mix with the crushed wheat grains. Then the yeast (Aspergillus oryzae or Aspergillus sojae) will be added to the mixture of steam soybean and crushed wheat grains. The ratio of wheat and soybean may vary depending on the type of soy sauce to be prepared. In order to activate the yeast, the mixture will allowed maturing for 2 or 3 days in a large of perorated vats in a control air circulated room. This process will generate heat and because of that it is good to check the temperature to prevent overheating that can kill the yeast. The right temperature of the mixture is around 30C to 31C. After the incubation, the Koji becomes greenish yellow as a result of mold growth and sporulation.

2. Brine fermentation

Figure 9: Brine Fermentation

The second step in making fermented soy sauce is brine fermentation. It utilizes the lactic bacterium, Pediococcus halophiIus and the yeasts Zygosaccharomyces rouxii and Candida species both of which tolerate a salt concentration of 20 g per 100 ml. The brine effectively prevents growth of undesirable microorganisms. The harvested Koji is mixed with 20% salt brine and water, and transferred by means of a spiral pump into deep fermentation steel tanks coated with epoxy resins on the interior. The resultant mixture is called moromi mash. The moromi will be blend for a few minutes and the tank will be seal in order for fermentation process occur. It is important to control the microorganisms in the brine fermentation. The specially selected Pediococcus halophilus is cultured and added to the mash. To control its 9

growth rate it is necessary to keep the fermenting mixture at 15 C for the first month, allowing the pH of the mash to decrease slowly from 6.5 to 5.0. Then cultures of Z rouxii and Candida species are added as a starter. The temperature of the moromi is allowed to rise slowly to nearly 28 C until vigorous alcoholic fermentation starts. The temperature in the tank can be controlled by coil type heat exchangers with mixing devices, thermocouples, and control systems in which controlled the tank to be open and mixing the moromi once a day for the first 6 weeks in order to remove the gases that build up and to stimulate yeast growth, mixes the contents and at the same time developing the flavour. During the fermentation period, proteolytic enzymes from Koji hydrolyze the proteins in soy bean and wheat to form amino acids and low molecular weight peptides. Starch is converted to simple sugars which are fermented primarily to lactic acid, ethanol and carbon dioxide. During the brine fermentation, the pH of the mixture drops from 6.5 to 5.0 in the first month at 15 C .This is followed by fermentation at 28 C for four months. The mixture will be open once a week for mix the moromi. After 6 month fermentation, the tank will be open and the moromi will be mix for the last time and it finally ready for next process. This fermentation process creates over 200 different flavours.

3. Refinement

and

Pressing

10

Figure 10: Refinement and Pressing The third process which and is the The

refining pressing

including

filtering.

aged moromi will be removed from the tank and it will be placed on the filtration cloth. Than the moromi that on the filtration cloth will be fold and place on the equipment for pressing process. During pressing process, moromi is placed into special equipment where the mash is strained through filtration cloth and pressed vertically to separate soy sauce from the residue. The moromi will be give 6 tan of pressure under the force of gravity and the moromi then will mechanically pressed slowly and steadily for about ten hours. It takes a considerable period of time to gradually press the mash in order to get delightfully clear and plain soy sauce. After pressing, the filtered raw soy sauce is pasteurized in a heat-exchanger at 70-80 ~ for a few minutes to ensure clarity, to inactivate residual enzymes, and to inactivate any undesirable microorganisms.

4. Pasteurization The soy sauce that has been pressed from moromi is known as raw say sauce. Raw soy sauce will be left in a clarifier tank for about 3 to 4 days to separate the raw say sauce into various components that is oil floating on the surface and sediment settling on the bottom. This oil can be reuse as a fuel for machinery operation and the sediments can be reuse for livestock feed. The clarified soy sauce will be undergoing for pasteurization process. This clarified soy sauce 11

will be run at steam pipe at temperature of 80C. There will be three

purpose of the

pasteurization process that is halts the activity of enzyme to stabilize the quality of soy sauce, prolong the shelf life of soy sauce and at the same time to adjust colour, taste and smell. Finally the pasteurized soy sauce will be bottle automatically.

2.1.2 FLOWCHART OF BREWING METHOD

12

Figure 11: Brewing Method Flowchart 1

13

Figure 12: Brewing Method Flowchart 2 14

2.2 NON BREWED METHOD (HYDROLISIS) There are some of soy sauce are been made by using chemical known as hydrolysis process. This process is different from brewed process and it much faster. Using a hydrolysis process only takes a few day compare to brewed method that take several months in order to produce soy sauce. Below are the steps for making a soy sauce using hydrolysis process; 1. Soybean has been boiled in hydrochloric acid for about 15 to 20 hours in order to remove the amino acid. 2. When the maximum amount of acid amino has been removing, the mixture will be cold to stop the hydrolytic reaction. 3. The mixture will be neutralized by using sodium carbonate. 4. Then the mixture will be pressed through the filter and mix with the active carbon for purified though the filtration. This solution is called as hydrolyzed vegetable protein. The result of this process is dark colour liquid and produces the glutamic acid that provides the flavour to the soy sauce. 5. The caramel colour, corn syrup and salt are added to this mixture in order to get the right colour and taste. Than the mixture will be refined and package. The soy sauce that are produce from chemical method are hasher and did not have pleasant flavour and character compare to those soy sauce that are made from brewed method. The difference is the flavour because the used of hydrolysis acids that tends to be more complete than fermentation counterpart. This means almost all protein in non brewed method are convert into amino acid while for brewed method the amino acid will stay as a peptides that will give a different flavour. Besides flavour, the non-brewed soy sauce is generally different with brewed soy sauce because it is dark in colour. Furthermore, it has a harsh, overpowering flavour and a pronounced chemical aroma. The production of chemical soy sauce usually only takes a few weeks because there is no natural brewing process. The soy protein is not separated by enzymes, as it is in the natural brewing process, but with hydrochloric acid. Colour, flavour and aroma are not produced naturally, but in the form of additives such as corn syrup, salt and caramel colour. The quality of the resulting soy sauce is therefore much inferior to that of Kikkoman naturally brewed soy sauce. 15

2.3 INGREDIENTS IN MAKING SOY SAUCE Certain soy sauce has different additives included during the process of making the soy sauce to give them special characteristic such as flavour and aroma. Below are the essential ingredients that need to be presence in order to produce soy sauce. Soybean Barley grains Salt Water Yeast Preservatives and other additives

2.3.1 EFFECT OF THE INGREDIENTS Soy bean 16

Figure 13: Soybeans

Soybeans have been known as King of Legumes because of precious nutritive contain in soybeans. Soybean has lowest in starch and high in protein. Moreover soybean also contain of high in minerals, particular calcium, magnesium and vitamin B. Soybean have variety of colour from yellow, black and green. The main ingredients in making a soy sauce is soybean and it will be mixing and mashed with other ingredients. The type of soy sauce is based from the protein contain inside the soybean. The protein in soybeans changes to components that produce flavour and colour unique to soy sauce as they are acted upon by microorganisms. Soybean will be soaking in water for certain period of time and it will be steam in high temperature. In soy sauce production, they use almost 70% of soy bean in the sauce.

Barley grains

Figure 14: Barley grains Barley is a cereal grain that was originally native to Asia. Barley was very important in ancient times and was one of the first grains to be widely cultivated. However, when leavened bread became popular, consumption of barley was replaced by wheat and rye. Barley grains contain high in carbohydrates. In making a soy sauce barley grains will blend and the crushed barley grains will be mix together with the soybean. Carbohydrates that contain in the barley grains are 17

the element that provide soy sauce with the pleasant smell and also give sweetness to the soy sauce.

Salt

Figure 15: Salt Salt will be added during the beginning fermentation process and give around 12-18% of the weight for finished product. The salt is added in the form of salt water and the salt does more than to add salty taste to the soy sauce. The salt helps to protect the soy sauce from spoilage because of high salt concentration. In making soy sauce, the salt not only has a function for flavouring but at the same time it a gives suitable chemical surroundings for lactic acid bacteria and yeast to do fermentation process properly. The salt protects Moromi from unnecessary microorganisms and helps slow activities of lactic acid bacilli and ferment, which are indispensable in soy sauce production. For a salty soy sauce, the amount of salt will be greater than the amount of salt in the sweet soy sauce.

Yeast

18

Figure 16: Yeast Microorganisms have been used by human to enrich eating habits. Yeast is one of the microorganisms that have been used in food production such as alcohol beverage, soy sauce, bakery item and etc. In soy sauce production, the mixture of barley grains and soybean exposed to specific mold that called Aspergillus Oryzae or Aspergillus Soyae. This kind of mold has been used to ferment soybean. Aspergillus Oryzae will breakdown the protein and carbohydrate to foaming a koji. Than the yeast and saltwater will be combine to form of moromi. Fermentation happened when additional bacteria called Lactobaccillus and yeast which enzymatically react with protein to produce a number of amino acid and peptides where all these protein will contribute to the flavour at the soy sauce.

Preservatives and other additives

Food preservation is the process of treating and handling food to stop or slow down the spoilage and thus allow for longer the shelf life of any products. Preservation usually involves preventing the growth of bacteria, yeasts, fungi, and other micro-organisms as well as retarding the oxidation of fats which cause rancidity Besides that, preservative food additives can be used alone or in conjunction with other methods of food preservation. For non brewed method in processing a soy sauce, additional colour and flavour agent have been added to give desirable colour and flavour to the soy sauce. The food preservatives also will be added in the soy sauce in order to make the sauce become high in quality and longer the shelf life of the products.

3.0 CHEMICAL COMPOSITION OF SOY SAUCE

19

In general, good quality soy sauce contains 1.0-1.65% total nitrogen (w/v), 2-5% reducing sugars, 1-2% organic acids, 2.0-2.5% ethanol and 17-19% sodium chloride (w/v). About 45% of the total nitrogen is found in simple peptides, and 45% in amino acids.

Table 1: Composition of soy sauce The properties of some Japanese soy sauces are shown in Table 1. Koikuchi is a representative of the fermented soy sauce popular in Japan. It is an all-purpose seasoning characterized by a strong aroma, attractive flavor, and a deep reddish brown color. More than 56% of Japanese soy sauces are of this type. The 'Upper grade' or 'Usukuchi shuyu' is used mainly in the western part of Japan. The 'Standard grade' or 'Tamari shuyu' sauce is characterized by higher total nitrogen and formol nitrogen compared with the other varieties. It is consumed largely in the Nagoya region of Japan. The Saishikomi and Shiro soy sauces are produced and consumed only in isolated localities or for special industrial uses. The amino acid composition of the soy bean, wheat and a representative Koikuchi soy sauce is presented in Table 2. Glutamic and aspartic acids are the major amino acids present in the fermented soy sauce. A total of 18 amino acids are present in the sample. Arginine, lysine, phenylalanine, serine, threonine, leucine, isoleucine, valine, alanine and proline are present in significant quantities in the sauce. A comparison of the amino acids present in the fermented soy sauce with those in the soy bean and wheat indicates the following changes: a) arginine is converted into ornithine in the fermentation process; b) tryptophan and cystine may have been used during the fermentation as nutrients; and c) tyrosine decreases in the moromi mash. 20

A representative Japanese fermented soy sauce contains the following sugars % (w/w): arabinose, 0.08%; glucose, 2.05%; mannose 0.06%; galactose, 0.17%; xylose, 0.06%; disaccharide, 0.65%; polysaccharide, 1.15%; unidentified sugar 0.23%; total sugar, 4.45% (as glucose). Organic acids: acetic acid, 0.16%; citric acid, 0.04%; formic acid 0.02%; lactic acid, 0.68%; succinic acid, 0.05%; total 0.95%.

Table 2: Distribution in Koikuchi soy sauce

3.1 FLAVOR COMPOUND

21

Based on distillation-extraction techniques and using various types of glass and fused silica capillary columns, it is reported that there are more than 200 peaks in the chromatograms which indicate the volatile compounds. These volatile compounds included: 1) hydrocarbons, 2) alcohols, 3) esters, 4) aldehydes, 5) acetals, 6) ketones, 7) furans, 8) lactones, 9) furanones, 10) pyrones, and 11) pyrazines. Of these aromatic compounds, HEMF (4-hydroxy- 2 (or 5)ethyl-5 (or 2) methyl-3-(2H)-furanone) appears to be a very important flavor component of fermented soy sauce. HEMF possesses a strong flavor and resembles the aroma of the moromi mash of Koikuchi soy sauce. The volatiles are formed by microorganisms during the Koji and brine fermentation as well as the heating and pasteurization process. There is also a report which reported on the quantitative aspects of 12 major flavor components in Japanese soy sauces as follows: isobutyl alcohol, 3.07-18.35 p.p.m.; n-butyl alcohol, 1.41-11.48p.p.m.; isoamyl alcohol, 4.47-22.0p.p.m.; acetoin, 5.08 8.44p.p.m.; ethyl lactate, 7.35-27.12p.p.m.; furfuryl alcohol, 4.35-10.07 p.p.m.; methionol, 2.60- 4.47 p.p.m.; 2-phenylethanol, 3.71-10.25 p.p.m.; HDMF (4- hydroxy-2,5-dimethyl-3(2H) furanone), 1.83-5.39p.p.m.; 4- EG (4-ethyl-2-methoxyphenol(4-ethylguaiacol)), 1.12-3.67 p.p.m.; HEMF (4-hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl- 3(2H)-furanone), 177.8-418.7 p.p.m.; HMMF (4 hydroxy-5- methyl-3(2H)-furanone), 84.54-153.6 p.p.m.

4.0 CONCLUSION

22

In a nutshell, the soy sauce production method have evolved from the conventional method to more sophisticated method which involved machineries, techniques, processes and additives that helps to preserve the taste and quality of the soy sauce. Innovations are made in producing varieties of soy sauce product that is available in the market but the main purpose is remaining the same, to serve the customer the best quality and safe soy sauce product to be enjoyed. Even though the process of making soy sauce is very hard and complicated even with the presence of technology, but the effort to preserve the taste of the soy sauce deserve a compliment. As most Asia Pacific and South East Asia countries used soy sauce in their traditional cuisines, the effort by Japanese Agricultural Standard (JAS) in grading the soy sauce should be exemplary and followed by others in which would be very helpful for customer to choose which soy sauce is the best and fits for them. The Halal certification should also be a benchmark for all soy sauce manufacturers and labelled and declaring all their product ingredients so that it will not raise any morals and religious codes especially for Muslim as they may find it unacceptable to consume non-halal food or indeed containing any ingredient that not declared. Undeclared ingredients used can have ramifications for minority groups such as vegetarians, vegans, pregnant women, children and anyone on special diets.

23