Tane koje and starter cake

1. Microbiology andTechnology of Fermented Foods
Graduate Program in Biotechnology
School of Life Sciences andTechnology

2.  Tane koji, means seed koji, is a spore mass of koji mould
produced in rice grains.Tane-Koji is also called Moyashi.
'Moyashi' means 'sprout’
 Tane koji is an important aspect in producing koji for
Japanese food fermentation such as Sake, Miso, Soy
sauce, Sochu, etc.
 Tane koji has been produced commercially in Japan, as
inocula in koji preparation for a variety of Japanese
fermented food since 14th century

3.  Tane-Koji is a starter culture of Asp. oryzae for making
Koji. Koji fungi are cultured on steamed rice at
polishing ratio 96% with trace of ash at 34 to 36℃ for
one week during the production ofTane-Koji.
 The ash is made by burning leaves such as camellia,
zelkove or oak in the roster.Ash are rich in minerals
such as potassium, magnesium and phosphate.
 The effects of ash are to increase spore yield and
survival rate, and to inhibit various contaminants.

4. Some differences in makingTane-Koji and Koji.
• The main purpose of Koji making is producing enzyme. However,
that ofTane-Koji is producing spores for Koji making.
• For Koji making, polished rice or barley is used and it takes about
two days. But forTane-Koji making, nutritious rice with high
polishing rate (brown rice) is used and it takes about one week.
• Tane-Koji is normally dried for quarity keeping.
• InTane-Koji making, about one parcent of ash is added to steamed
and cooled rice to improve growth and spore forming capacity of
Koji fungi, and also to improve preservation. The best ash is
camellia ash, but sometimesTane-Koji is made without ash.

5.  Several kinds ofTane-Koji have been developed
from different strain of Aspergillus that have been
handed down as heirlooms by every producers and
some of them are obtained by natural or artificial
mutation.

10. Starter cake composed of mixed cultures of
filamentous fungi, comprising mostly the Mucorales
and yeast  used as inocula in fermentation of
starchy raw materials into sweet alcoholic foods
(sweetened rice) or alcoholic beverages (rice wine)
The technology concerning preparation of these
inocula is believed to have originated in China.The
product is called by various names in Chinese such as
chiu-yueh, peh-yueh, chi-chu, etc.
Similar starter cakes produced in otherAsian
countries are called by various local names

11. Rice or wheat is
ground and mixed
with certain spices
Water is added to
make a dough-like
material
The dough is shaped
(usully small ball) and
placed on a wooden
tray
Powder from an old
starter is sprinkled over
the ball, and tray is
covered with cloth
Incubated in room
temperature for 2-5
days
Drying in the sun,
and cakes can be
kept for many
months

12. Spices used in starter cake recipes:
• As selective inhibitors, preventing growth of
undesirable microorganisms
• Various spices has been used, most producers
regard their recipes as trade secrets passed
from generation to generation
• In general, a mixture of garlic, pepper, rhizomes
of Alpinia galanga and Capsicum frutescens is
used in preparation of the Indonesian ragi

13. LOOG-PANG KAOMAG
(sweetened rice)
 Allium sativum 420 g
 Zingiber officinale 120 g
 Glycyrrhiza galabra 180 g
 Piper nigrum 60 g
 Piper chaba 120 g
 Alpinia galanga 60 g
 Rice 12kg
LOOG-PANG LAO
(Thai rice wine)
 Allium sativum 40 g
 Zingiber officinale 40 g
 Alpinia sianensis 20 g
 Myriopteron extensum 40 g
 Piper nigrum 6 g
 Piper chaba 6 g
 Allium ascalonicum 20 g
 Rice 2,5 kg

14. Local Name Country Product
Chiu-Yueh, peh-yueh China Sweetened rice
Chi-chu China Rice wine, spirit
Bukhar, murcha India Rice wine
Nuruk, chi-chu Korea Spirit
Ragi tape Indonesia Sweetened rice
Ragi peuyeum Indonesia Sweetened cassava
Ragi tempe Indonesia Tempe
Ragi tapai, jupiang Malaysia Rice wine
Loog-pang kao-mag Thailand Sweetened rice
Loog-pang lao Thailand Spirit, rice wine
Bahn men Vietnam Rice wine, sweetened rice
Bubod Philippines Fermented toddy

15. Numerous filamentous fungi and yeasts have been isolated
from starter cakes from various sources.
Among the moulds,
Amylomyces rouxii
(Clamydomucor oryzae),
Rhizopus spp., Mucor
spp., are those isolated
from most of the
samples
Hansenula malanga and
Endomycopsis fibuligera
are the yeasts common
in many starter cakes.
Lactobacilli and the
cocoid lactic acid
bacteria have been
isolated from some
samples of starter cakes
Among microorganisms isolated from various sources, only certain genera
are found to play leading roles in the fermentation of sweetened rice and rice
wine: Amylomyces rouxii, Rhizopus spp., Endomycopsis spp., Hansenula spp.,
and Saccharomyces spp

16. Starter cake Molds
Chinese yeasts Rhizopus javanicus, R. chinensis,
Amylomyces rouxii
Ragi Amy.rouxii, Mucor dubius, M. javanicum, R.
pryzae, Aspergillus niger, R. stolonifer, M.
rouxii, R. cohnii, Zygorrhynchus moelleri, A.
oryzae, A. flavus, R. oligosporus, R. arrhizus,
Fusarium spp.
Loog-pang Amy.rouxii, Mucor spp., Rhizopus spp., A.
oryzae
Murcha M. fragilis, M. rouxii, R. arrhizus
Bubod Rhizopus spp., Mucor spp.

17. Starter cake Yeasts
Chinese yeast Endomycopsis spp
Ragi Torula indica, Hansenula anomala,
Saccharomyces cerevisiae, Endomycopsis
chodati, E. fibuligera, H. subpellliculosa,
H.malanga, Candida guiliermondii, C.
humicola, C. intermedia, C. japonica, C.
pelliculosa
Loog-pang Endomycopsis fibuligera
Murcha H. anomala
Bubod Endomycopsis spp., Saccharomyces spp.

18. Tane koji and starter cakes are similar in usage
The difference between koji and starter cake:
• Microorganisms involved:Tane koji mostly consist of
A.oryzae, A.sojae or A.awamori , whereas starter cakes
usually consist of mixed culture of moulds of the
Mucorales and yeasts
• Materials: koji molds grown on cooked rice grains,
whereas mixed culture of starter cake grown on raw rice
meal

19. Koji process has been applied to almost all solid
substrate fermentation
The term Koji derives from the chinese character,
meaning moldy grains
• Shui in Chinese
• Koji in Japanese
• Ku in Korean
Pronounced differently
in the various
languages deriving
from Chinese

20. Koji can be name according to
The cereal used
• Kome (rice) Koji
• Muji (barley) Koji
The colours of
spores/pigments
produced
• Ki-Koji (yellow) : A.
oryzae
• Kuro-Koji (black) : A.
awamori
• Beni-Koji (red) :
Monascus purpureus
The end use of
Koji
• Miso Koji
• Shoyu Koji
• Sake Koji

21. • Solid substrate cultivation of molds to produce
hydrolytic enzymes
• Koji provides a source of enzymes necessary to
convert solid, raw materials containing complex
substrates into soluble simple products that can be
easily fermented
• Various chemicals essential for the 2nd fermentation
stage or those imparting flavor to the products are
also produced
• The use of Koji is analogous to malt in beer
Koji preparation is essential step in fermentation of
variety of fermented food of the Orient

22. Raw material treatment is to enhance its digestibility,
enhances protein extraction from raw material
Soybean
• Soaked for about 12
hr
• Cooked with steam
under pressure for 1
hr (puffing)
Wheat (soybean
koji + wheat)
• Roasting wheat
kernels
• Crushing
Rice (Rice koji)
• Polished rice (rice
minus the bran)
• Steamed

23. Should produce
proteolytic,
peptidolytic and
amylolytic enzymes
with high activity
Should sporulate and
grow well on their
substrate
Should not produce
toxins under
production condition
Should be genetically
stable and produce
products having
consistent flavor and
color properties

24. Inoculation of koji
substrates:
Inoculated with a pure culture
containing spores of fungi
(Aspergillus oryzae and/or A. sojae)
Inoculated with 0.1% to 0.2% of
seed culture (Tane Koji) that is
usually contains multiple strains

25. Koji substrate that has
been inoculated is
mixed and incubated in
large rectangular trays
or boxes
Perforations in the
trays enhance air and
moisture circulation
Temperature control is
important:
• Proteolytic enzyme: 25oC –
30oC
• Amylase: > 30oC – 35oC

26. Two-stage
fermentation
• Stage one: hydrolytic
enzyme production by
koji molds
• Stage two:
fermentation stage
• Sake brewing, shoyu,
miso, etc.
Single-stage
fermentation
• Tempe: Rhizopus
oligosporus
• Tempe bongkrek:
Rhizopus oligosporus
• Oncom: Neurospora
sitophila
• Sufu: Mucor