TEMPEH

Traditional Fermented Food

of Indonesia

Assoc. Prof. Le Hong Phu

Tran Minh Quan BTFTIU16080

Dinh Yen Nhi BTFTIU17080
Tran Huynh Phuong Khanh BTFTIU17056
Trinh Quoc Khanh BTFTIU18188
 01 Introduction

02 Processing

03 Physicochemical changes
during fermentation
Outline
04 Effect of tempeh

05 Summary

06 References
1. Overview
• Tempeh is a mold-fermented soybean food. Occasionally other legumes, seeds,
and cereal grains are also used along with soybeans. The mold fermentation
results in a matrix of dense, cottony, mycelia in which cooked soybeans are
embedded, forming a compact greyish-white cake.

• Tempeh is a popular fermented food of Indonesia. However, in the last two

decades it has attracted the attention of the Western world, particularly
North America, because of its unique flavor, sliceable meat-like texture,
and nutritional attributes.
• It is free of cholesterol, a good source of vitamins and minerals, and highly digestible,
and is also considered to impart growth-stimulating effects.
• It can serve as an excellent substitute for animal protein products such as meat,
chicken, fish, etc. Fresh tempeh contains approximately 19-20% protein, which
compares well with the protein content in chicken (21 %), beef (20%), hamburger
(13%), whole eggs (13%), and whole milk (3%).

Table: Nutrient Composition of Tempeh (per 100 g) Table: Vitamin and Mineral Contents in Fresh Tempeh
Dishes with Tempeh

Tempeh taco
Teriyaki Tempeh Stir-fried Tempeh

Tempeh BBQ

Baked Tempeh Tempeh curry

Fried Tempeh Tempeh burger
2. Production of Tempeh

• There are different scale of tempeh production: home scale, small cottage
industries, larger commercial processing plant
• High protein plant sources are used as alternative to animal sources through
solid state fermentation (SFF)
• Fermentations producing a meat-like texture in a cereal grain or legume
substrate by means of a fungal mycelium that knits the particles together
• Ingredient: Beans or grains, mold, container,
 Step of Tempeh processing

Wrapping and
Cleaning Innoculation
incubation

Draining and
Dehulling Packaging
cooling

Hydration
and Acid Cooking
fermentation
• Disadvantage of small home scale: high total solid loss 22 to27%.
Which is caused by dehulling (8 to 17%), soaking and cooking (6
to 12%). and fermentation (1 to 4%)

The reduction of dry matter losses and required processing

energy is a challenge for food technologists.

• New processes are proposed involving dry technologies and

reduced cooking periods to ensure higher tempeh yields and less
waste.
2.1 Cleaning

The first step in preparation for the production of tempeh.

This step is carried out in order to remove dirt, stones, weed, seeds, damaged and
possibly decomposed beans, and any other foreign matter.
 Only the whole and healthy bean were selected
 2.2 Dehulling
Dehulling is the process of removing the hulls, outer
coverings or husk from beans and other seeds. 
 Dehulling is essential in the production of tempeh
as the mold R. Oligosporus cannot grow on whole
soybeans with hulls
 Dehulling enables the mold to reach the
nutrients in the cotyledons.
2 types of duhulling: wet and dry dehulling.
2.2 Dehulling

Wet dehulling
+ Generally done after precooking, which facilitates hydration of the soybeans.
+ Requires no mechanical devices other than the hands or feet to rub the hulls
from the cotyledons.
+ Thus, this method may not be suitable for large-scale tempeh production
2.2 Dehulling
Dry dehulling
+ The soybeans are heated for 10 min at 93°C to shrivel
the cotyledons and loosen the seed coats.
+ Dry dehulling is performed before any hydration
procedure and is a desirable, efficient method providing that
suitable mechanical equipment is available( burr, corn, or
steel roller mills may be used to crack the hulls).
+ Following dry dehulling, the hulls can be separated
from the cotyledons by the use of an aspirator, a gravity
separator, or by a winnowing process.

 Applicable for large scale production

2.3 Hydration and Acid fermentation

• Hydration is a process whereby soybeans are soaked in excess water for 12 to

15 h at room temperature in order to facilitate mycelial penetration.
• Under natural conditions, bacterial fermentation occurs during hydration
(soaking) and results in acidification.
• Soak water may be acidified artifically using lactic or acetic acid. Lactic acid
(50.5%) or acetic acid (10.25%) was reported to control microbial spoilage when
added during hydration.
• Lowering the initial pH allows for the mold to grow, while bacterial growth is
suppressed  the mold is not inhibited until the pH falls below 3.5.
2.4 Partial cooking

• Necessary to destroy contaminating bacteria that could interfere with

fermentation, to destroy antinutritional factors such as trypsin and
chymotrypsin inhibitors and to release some of the nutrients required for mold
growth.
• Boiling in excess water serves the purpose of partial cooking, which facilitates
fungal penetration and human digestion.
• Traditional cooking times varied from 10 minute to 3h. Cooking via steaming for
about 30 min at 100°C also has been reported as an acceptable method of
partial cooking.
• The bitter soybean taste disappeared in 115 min at 95°C. During the boiling
process, a heat-stable, water-soluble mold inhibitor is discarded as leaching out
into the water.
2.5 Draining, Cooling and Surface drying

• The boiling water is drained and the cotyledons are spread onto plated
bamboo trays, a wire mesh or woven basket was used to drain the soy beans.
• Excess water favors bacterial growth and spoilage of tempeh
Improperly drained soybeans decrease the shelf life of tempeh.
• The soybeans should be cooled to 38°C prior to inoculation. Cooled cotyledons
should be dull in appearance, indicative of a relatively dry surface. The
Malaysians surface dry the soybeans by rolling them in a piece of cloth, whereas
other manufacturers coat the beans with wheat flour.
2.6 Inoculation
• Indonesians used small pieces of soybean tempeh from a previous batch to
inoculate soybeans.
• While in the U.S., pure culture inoculation is used to make tempeh. Pure culture
inocula of Rhizopus spores are usually freeze-dried or suspended in water.
• Excess inoculum promoted rapid and uniform fermentation, if too little inoculum
was used, bacteria would be allowed to grow.
• Fermentation failures and excessive heat production were reported to be caused
by insufficient packing density with pockets of air and heavy inoculation.
2.6 Inoculation

• R. Oligosporus is the principal species of mold used to make tempeh because

the mold perform strong protease and lipase activity.
• Rhizopus strain used for tempeh production should have the following
characteristics:
1. Rapid growth at 37°C
2. High lipolytic activity
3. Production of strong antioxidant activity
4. Inability to ferment sucrose
5. Ability to produce the typical tempeh flavor, aroma, and texture
6. High proteolytic activity, resulting in the release of free ammonia after 48 to
72 h of fermentation.
2.6 Inoculation

• Other species such as R. Oryzae, R. Chinensis and R. Arrhizus also are used in
soybean tempeh production.
• R. Arrhizus to produce broad bean tempeh with the best flavor, texture, and
nutritional value. While R. Chinensis and R. Cohnii produced high quality
soybean and winged-bean tempeh.
• Other qualities, such as biosynthesis of B vitamins and inhibition of the growth,
sporulation and production of alfatoxin, also were reported as desirable
characteristics.

Broad bean
Winged bean
2.7 Fermentation containers

Wilted banana leaves or other large leaves

such as golden berry or tropical almond
leaves were reported to serve as excellent
wrapping for traditionally fermented
tempeh.
 Keep the soybean cotyledons moist
during fermentation
 Allow for gaseous exchange to occur.
2.7 Fermentation containers

• Some small factories used covered stainless steel pans or dryer trays
to ferment tempeh. These trays were lined with waxpaper to help
the soybeans maintain moisture and to prevent excessive aeration,
which would lead to sporulation and darkening of the mycelium.

• Perforation in lastic bags and plastic tubes at 0.25- to 1.3-cm intervals

to allow access of oxygen exchange for the mold. Plastic bags were
successfully adopted for use in tempeh manufacture in Indonesia.
2.7 Fermentation containers

• Leaves, plastic, glass, wood or stainless steel can serve as appropriate container
material as long as certain criteria are met.
• A fermentation container is appropriate as long as it permits access of sufficient
oxygen for the mold's growtah, but not too much to promote sporulation and
darkening of the mycelium
• Temperature can be controlled, the soybeans maintain moistness during
fermentation, there is no free water in contact with the soybeans, and the
fermenting tempeh remains clean and wholesome.
2.8 Incubation
• Temperature, length of fermentation and relative humidity are three crucial
factors that dictate the outcome of tempeh fermentation.
• Tempeh fermentation can be carried out at temperatures ranging from 25 to
37°C. The time required for fermentation decreases as the temperature
increases.
• Higher temperatures (up to about 37oC) influenced mold growth and led to a
shorter incubation time. The temperature within the fermenting beans should
not exceed 42°C because growth of the mold would be hindered.
 Conditions for producing high quality tempeh are quite variable. Temperature,
relative humidity, fermentation length, oxygen availability, and other conditions
must guarantee that the overall growth requirements of the mold are met.
 With proper control of these conditions, R. oligosporus grows at a rapid rate,
yielding tempeh in a minimum length of time.
2.9 Harvesting, Storage and Preservation

• Tempeh should be harvested after the soybean

cotyledons have been overgrown with mold and have
knitted into a compact cake. Furthermore, the
cotyledons should feel soft and pasty (not rubbery) when
pressed between the fingers.
• Freshly made tempeh can be stored for several days at
room temperature without adversely affecting the
nutritional or organoleptic properties.
• Storage stability of tempeh can be extended by drying,
frying, dehydration, freezing, canning, steaming,
blanching and other preservation methods.
 3. Physicochemical changes during fermentation
• The neutral fat of soybeans is composed of palmitic, stearic, oleic, linoleic, and
linolenic acids. Enzyme produced during fermentation is strongly lipolytic and
hydrolyzes one third of the fat during fermentation at 37 °C.
 The acid content of soybeans increases approximately 30- to 50-fold during
tempeh fermentation.

• Protein content in tempeh increased 16% on average in relation to that of

soybean grains
Protein content increased in tempeh due to the cooking and fermentation steps
during tempeh processing, which favor losses of soluble substances (mineral and
sugars) from soybean grains.
3. Physicochemical changes during fermentation
• Isoflavone content of tempeh double to those in grain before fermentation
isoflavone aglycons are not formed in fresh harvested soybean grains

Fermentation or moisture activates the β-glycosidase enzyme hydrolyzing

isoflavone glycosides to form aglycon isoflavones

 Isoflavone, primarily their aglycones (genistein and daidzein) involved in a

number of biological activities including breast and prostate cancer chemo-
preventive activity, and the ability to modify carcinogenesis,

• The content of Kunitz trypsin inhibitor was remarkable reduced in tempeh.

Cooking, soaking, and dehulling all contribute to the reduction of trypsin
inhibitor and reduce about 80% of the trypsin inhibitor activity in the soybean
grains
 enhance nutritional value of tempeh
3. Physicochemical changes during fermentation

Traditional tempeh fermentations contain vitamin B, whereas tempeh made with

pure mold culture under aseptic conditions was found to lack vitamin B,
identified the bacterium responsible for vitamin B, activity as Klebsiella pneumoniae,
which is a common inhabitant of plant material.
 K.pneurnoniae becomes essential for tempeh fermentation in order for tempeh to
provide vitamin
  Rich in nutrients
4. Effect of
Tempeh on health  Provide prebiotics

 Support digestion

 Promote bone health

Benefits
 Decrease oxidative

stress

 Bloating
Non-benefits
 Soy allergy

 Peptic ulcer
5. Summary

 An important function of microorganisms involved in the fermentation process is

the synthesis of enzymes that hydrolyze the food ingredients and contribute to the
development of a desirable texture, flavor and aroma of product.

 Fermentation also decreases antinutritional constituents, and the nutritional

quality and digestibility of the fermented product is improved.

 Fermented foods could enhance the barrier function of the gastrointestinal tract,
possibly resulting in reduced diarrheal disease risks.
 Fermented soybean foods being nutritious, easily digested and absorbed,
culturally acceptable, palatable and possibly protective against diarrhea might break
the viscous cycle of diarrhea and malnutrition.
 Futhermore, the present or potential role of fermented soybean foods in
prevention and treatment of chronic diseases merits increasing attention.
6. References

• B. K. Mital, S. K. Garg, G. B. Pant, 1990. Tempeh—Technology and food value.

Food Reviews International, 6(2), 213-224.
• Kathleen A. Hachmeister and Daniel Y. C. Fung, 1993. Tempeh: A Mold-Modified
Indigenous Fermented Food Made from Soybeans ,and/or Cereal Grains. Critical
Reviews in Microbiology, 19(3):137-188.
• Miladinovic, Jegor & Djordjevic, Vuk & Balesevic Tubic, Svetlana & Petrović,
Kristina & Ceran, Marina & Cvejic, Jelena & Bursać, Mira & Miladinović, Dragana.
(2019). Author Correction: Increase of isoflavones in the aglycone form in
soybeans by targeted crossings of cultivated breeding material. Scientific Reports.
9. 17137. 10.1038/s41598-019-53778-y.
• Bavia, Ana & Silva, Carlos & Ferreira, Márcia & Leite, Rodrigo & Mandarino, José
& Carrão-Panizzi, Mercedes. (2012). Chemical composition of tempeh from
soybean cultivars specially developed for human consumption. Food Science and
Technology (Campinas). 32. 613-620. 10.1590/S0101-20612012005000085.
Thank you very much for
your attention