FOOD FERMENTATION TECHNOLOGY

NON-SOYBEAN TEMPEH FERMENTATION

QURRATA A’YUNI F24160069

MICHAEL OSCARIUS F24160081
MARTHA ULI SINAGA F24160088
M.ALDY MUSTOFA F24160019
ADELE MARVAUD F24188525

FOOD SCIENCE AND TECHNOLOGY

FACULTY OF AGRICULTURAL TECHNOLOGY
IPB UNIVERSITY
2019
 ABSTRACT
Tempe is a fermented food product by several microorganisms. Some of
them are Rhizopus oligosporus, Rhizopus oryzae, which are molds, LABs, and
bacteria such as Bacillus sp and Klebsiella pneumoniae. Bogor beans represents
an interesting source of raw material to make tempeh, it could promote a local
ingredient and have economical impact. It contains the nutritional content for
every 100 g of Bogor bean seeds includes: total carbohydrates 61.7%, fat 6.3%,
protein 17.7%, fiber 4.9%, ash 3.3%, thiamin 0.28 mg / 100g, riboflavin 0.12 mg /
100 g, calcium 73 mg / 100 g, iron 7.6 mg / 100 g, vitamin A 30 iu / 100 g, and
phosphorus 0.38 mg / 100 g (Hamid 2009). The principles of tempe processing
using Bogor beans are similiar to the common tempeh processing. Those are
washing, boiling, soaking, washing, adding of inoculum, packaging, and
fermentation. The fermentation process result the chemical and physical changes
of Bogor beans, and compact texture of tempeh because of micelial growth
allover the beans.

Keywords: Bogor beans, tempeh, fermentation

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 TABLE OF CONTENTS

TABLE OF CONTENTS ii

INTRODUCTION 4

Background 5

Purpose 5

LITERATURE REVIEW 6

The availability of Bogor beans 6

Bogor beans Processing 7

Result of Biochemical and Physical Changes caused by Fermentation 9

CLOSING 11

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 CHAPTER I

INTRODUCTION

Background

Soybeans have long been used as raw materials in making tempeh (Haliza
et al. 2007). Tempe provides an easily digestible source of protein, as well as
essential vitamin B12 for the body (Muchtadi 2010). Soybean production based
on BPS data in 2012 only reached 843.15 (in thousands of tons), while Indonesia's
national needs reached 2.3 million tons per year, this caused the Indonesian
government to import soybean commodities.
Tempe is a fermented food product by several microorganisms. Some of
them are Rhizopus oligosporus, Rhizopus oryzae, which are molds, LABs, and
bacteria such as Bacillus and Klebsiella pneumoniae (Nuraida et al. 2008; Sukardi
et al. 2008). The growth of mold in soybeans forms biomass in the form of white
mycelium, the mold hydrolyzes the protein contained in soybeans into amino
acids and also converts large molecules such as fats and carbohydrates into simple
molecules such as fatty acids and glucose. In addition, tempeh also has isoflavone
content formed during the fermentation process which has an antioxidant function
(Muji et al 2011).
The limitations of soybeans as raw material for making soybeans make the
researchers think of doing research to make tempeh from various sources of
beans. Because in principle, making tempe is an increase in biomass by molds that
use protein from peanuts as nutrients to grow, this means that tempeh should be
made from sources of beans other than soybeans. Previous research has explored
the development of tempe products from green beans and kidney beans. The
results of the study generally indicate that tempeh made from red beans and green
beans has not been able to show potential as a raw material.
Bogor beans as one type of beans are quite abundant with production
reaching 4 tons per hectare. In other hand bogor beans more easily to culivate
because of its high temperature tolerance and low soil demands. results showed
that the nutritional content for every 100 g of Bogor bean seeds includes: total

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carbohydrates 61.7%, fat 6.3%, protein 17.7%, fiber 4.9%, ash 3.3%, thiamin 0.28
mg / 100g, riboflavin 0.12 mg / 100 g, calcium 73 mg / 100 g, iron 7.6 mg / 100 g,
vitamin A 30 iu / 100 g, and phosphorus 0.38 mg / 100 g (Hamid 2009). Because
of that high potential, Bogor beans is a potential sources to make tempeh.

Purpose

The purpose on making non-soy tempeh are to increase the utilization the
high potential of Bogor beans, decrease the utilization of soy as raw material of
tempeh, and increase the diversification of non-soy tempeh fermentation product.

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 CHAPTER II

LITERATURE REVIEW

The availability of Bogor beans

Tempe is a fermented food from Indonesia and made from soybeans. In

Indonesia, soybean production in 2012 was only 843.15 thousand tons, while
national demand was 2.3 million tons ([BPS] 2014). The high soybean needs
make the government import soybeans from other countries to meet the domestic
soybean needs. Indonesia has very large biodiversity, it should be able to give a
solution to overcome the needs of soybean as raw materials for making tempeh.
Some domestic beans has the potential to be developed into raw materials for
tempeh. Many types of beans can be used to make tempeh, including kidney
beans, mung beans, Bogor beans, and peanuts. (Radiati and Sumarto 2018). One
type of those beans that has not been widely used by the community is Bogor
bean.
Vigna subterranea (L.) Verdcourt, better known as Bogor beans, is one of the
type of legume less known in Indonesian market. This plant has been much
attention with a lot of research that reveals that beans Bogor is a promising food
source but is not very concerned. Utilization of Bogor beans that are not fully ripe
is directly for consumption while the dried ones are used to be processed into
flour (Astawan 2009).
Bogor beans allow it to be developed in semi-arid areas and nutrient-poor
soils. This plant can adapt well for short days. In the tropics, Bogor beans are
cultivated to an altitude of 1600 m above sea level. This plant live in a daily
temperature range of 20-28 ° C. Suitable rainfall conditions are 600-750 mm /
year. Optimal results are obtained on slightly sandy soils and have sufficient
calcium content with pH 5.0-6.5. In Indonesia, Bogor beans have long been well
adapted in the Bogor region and the eastern part of West Java, so that they are
better known as Bogor beans. At present, Bogor beans have spread to Sukabumi,
Majalengka, Tasikmalaya, Bandung, Central Java (Pati and Kudus), East Java
(Gresik), Lampung, NTB and NTT (Kuswanto et al 2012). The average yield of

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Bogor beans grown by farmers is still below 4 tons per hectare. Bogor beans are
harvested at the age of 115 - 120 days with the oldest leaf characteristics the
plants have begun to brownish and some dry out and the pods look full and the
seed coat is brownish. (Kurnia and Wahyudi 2018). By that quite high
productivity, Bogor beans represents an interesting source of raw material to
make tempeh.
Bogor Beans Tempeh Processing
Basically, non-soybean tempeh processing is developed based on the method
to make soybean tempeh. The principles of non-soybean tempe processing such as
washing, boiling, soaking, washing, adding of inoculum, packaging, and
fermentation (Handoyo and Morita 2006). The raw materials non soybean tempe
used Bogor beans as local ingredient from West Java. So in this process, no need
soaking for for a long time, cause the skin of Bogor beans easier to remove. The
method of Bogor beans processing such as:
● Boiling. This step aim to allow the beans to absorb as much water as
possible, till the Bogor beans were softer and easier for fermentation
process by acidification as a parameter in the initial stages. The ideal
boiling for non-soybean tempeh process is done twice to maximizing the
number of isoflavones as bioctive compound contained id tempeh. If
without boiling process in the initial stage, it can be used a longer soaking
time within acidic. The second boiling process is actually needed to ensure
that the beans are fully cooked and to kill contaminant bacteria. The
heating process likely boliling has a significant effect on decreased phytic
acid content of tempeh. The boiling process can also be decrease
allergenicity in Bogor beans. According to (Wilson et al. 2005) the
allergenicity of food can be influenced by the process of heating (include
boiling and steaming), fermentation, enzymatic hydrolysis, conjugation
with carbohydrates, and extrusion processes. The heating process of Bogor
beans allergen proteins at extreme temperatures can reduce IgE binding
capacity. Heating causes a decrease in the reactivity of allergenicity
(Sitorus 2014).

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● Stripping of Bogor beans shells. This process aims in order to lactic acid
to enter more easily into beans seeds and mycelium growth during
fermentation. Peeling peanuts on a small scale can be done using legs, but
if the beans are in large quantities using a peeling machine ((Handoyo and
Morita 2006).
● Soaking. This process aims for lactic acid fermentation to occur and the
occurrence of acidic conditions so as to encourage the growth of tempeh
mold, which will be achieved if the pH is around 3.5–5.2. The mixture of
peanut shells in tempeh will inhibit the growth of lactic acid bacteria
during soaking and reduce the acidification of Bogor beans. Bacterial
growth is characterized by the release of acidic odors during soaking and
the presence of foam on the surface of immersion water. Adding vinegar
to soaking water can accelerate the acidity process, even saving up to 10
hours of soaking time. Acidity and soaking also give a benefit bacterial
growth for the synthesis of vitamin B2, vitamin B6, vitamin B12, niacin,
biotin, folic acid, and pantothenic acid. In this study, soaking with vinegar
acid up to pH 4-5 for 7 hours ((Handoyo and Morita 2006).
● Washing. Aim that the beans do not become acidic and remove mucus
produced by lactic acid bacteria. The presence of bacteria and mucus will
hinder the final fermentation process. After washing, some producers boil
the beans a second time. This will make the beans softer. In addition, it
will also kill bacteria that live and multiply during immersion. Bogor
beans with two boiling times will be cleaner, longer storage, and not sour
Steaming was done to prevent damage to the texture of the beans
((Handoyo and Morita 2006).
● Cooling. Aim to cool before the beans are given yeast. While waiting for
the beans to cool, the beans are also cleaned of dirt that might still be
there. Dirt that is usually found in nuts is gravel, twigs, and other physical
contaminants (Handoyo and Morita 2006).
● Adding of Molds. The beans must be completely clean, dry and cold
before the yeast is spread on the surface of the beans. Some types of yeast
are mixed with rice flour or corn flour, the purpose of which is to facilitate

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 the yeast being spread in beans. R. oligosporus is the principal species
used in Indonesia for tempeh fermentation. One strain, R. oligosporus
NRRL 2710, is the recommended strain for producing tempeh
commercially. In addition, the strain for fermentation used other
microorganism such as Rhizopus oryzae, which are molds, LABs, and
bacteria such as Bacillus sp and Klebsiella pneumoniae to give a better
texture and flavour (Handoyo and Morita 2006).
● Packaging. After fermentation, the beans are immediately packed.
Packaging can use banana leaves or plastic with small holes to get oxygen
for mold growth. According to, the growth of molds from nuts wrapped in
leaves is generally faster than those wrapped in plastic. This packaging
part is used as treatment in the preliminary research. The research
treatment was carried out by comparing the differences in the use of
packaging types, namely banana leaves and polypropylene (PP) plastic
with different aeration (Handoyo and Morita 2006).
● Fermentation. After the packaging is complete, the beans that have been
wrapped, are squeezed in a place that is considered moist. The temperature
should not be too cold because it will hinder mold growth. The ideal
temperature ranges from 20 oC to 37 oC. The number of yeast, temperature
and humidity are important factors for the fermentation process. The
duration of fermentation is around 36 hours (Handoyo and Morita 2006).

Result of Microbiological, Chemical and Physical Changes caused by

Fermentation

During tempeh making there are many changes that occur in fermentation
process. Mainly this changes divided into three categories

a. Microbiological changes
Bogor bean tempeh incubatied for around 36-48 hours. In
microaerophyl condition mold, grow as micelia that cover bean. Rhizopus
oligosporus is main microbes that grow on tempeh. Other microbes like
Rhizopus oryzae and yeast usually grow and contribute to flavor of tempeh.

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 If incubation process took aver 48 hours, mold will sporulate and the spores
looks as black spot on tempeh.
b. Biochemical and physical changes
Molds and yeast growth will changes chemical compounds in bogor
beans. Rhizopus oligosporus produce enzymes that breakdown
macromolecules such as protein, lipid, and sugar (Tahir et.al 2018).
Proteolytic enzyme like protease will cut off protein into smaller peptides
molecules. Phytase enzyme degrade phytic acid that is tripsin inhibitor in raw
beans. This make tempeh have better protein biovaibility compared to other
protein sources. Some peptides contributes to flavor of tempeh. Sometime
bitter taste occur in tempeh because specific microbes has different
mechanism to fermented protein. According to some mold can release
antioxidant like isoflavone. Lipolytic activity of R. oligosporus 2710 on
showed in linoleic acid from hydro- lysed soybean lipids is reported to be one
of the main substrates used by growing Rhizopus as a molds to make a
tempeh (Radiati and Sumarto 2016). R. oligosporus has strong lipase activity
and therefore breaks beans lipids down into free fatty acids during
fermentation. Also observed in R. oryzae possessed strong lipase activity and
caused the hydrolysis of over one third of the neutral fat of the beans during a
3 day fermentation period. Of the free fatty acids that are liberated, only
linolenic acid was used by R. oligosporus. Glycerides in raw beans are broken
down into free fatty acids during the first 30 h of fermentation. Changes in
lipids during tempeh fermentation is identified the free fatty acids that were
liberated as palmitic, stearic, oleic, linoleic, and linolenic acids, with linoleic
acid predomi- nating (Handoyo and Morita 2006).
Bogor beans tempeh has compact texture because of micelial growth
allover the beans. This micelial gives tampeh its white unique color. Bogor
beans tempeh has moldy aroma but if the incubation process to long,
microbes produce secondary metabolites that sometime have unpleasant odor.

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 CHAPTER III
CONCLUSION
Bogor beans represents an interesting source of raw material to make
tempeh, it could promote a local ingredient and have economical impact.
Moreover it has technological interesting aspect, indeed the soaking step is
shorter. The micro-organisms used is similar to the one used for tempeh
fabrication, called Rhizopus oligosporus. Then lactic acid fermentation occurs,
and it needs specific conditions in order to kill the bacteria that could be pathogen
and will give also special flavour, more bitter. The characteristics of this kind of
tempeh are due to microbiological changes and chemical and physical as well.
The content of vitamins remains important such as vitamin 12 and others, and has
high bioavailability.

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 REFERENCES
[BPS] Badan Pusat Statistik. 2014. Produksi Tanaman Pangan 2014, Jakarta(ID): BPS.
Astawan M. 2009. Sehat dengan Hidangan Kacang-kacangan dan Biji-bijian. Depok
(ID): Penebar Swadaya.
Haliza, W., Y. Endang., Purwani, dan R. Thahir. 2007. Pemanfaatan Kacang –
kacangan Lokal sebagai Substitusi Bahan Baku Tempe dan Tahu. Buletin
Teknologi Pascapanen Pertanian. Vol. 3.
Hamid MN. 2009. Menggali potensi genetik tanaman kacang bogor (Vigna
subterranea (L.) Verdcourt). [skripsi]. Bogor(ID): Institut Pertanian Bogor.
Handoyo T, Morita, N. 2006. Structural and functional properties of fermented
soybean (Tempeh) by using Rhizopus oligosporus. International Journal
of Food Properties. 9(2), pp.347–355.
Kurnia R dan Wahyudi S. 2018. Pertumbuhan dan hasil kacang bogor (Vigna subterranea
(L.) Verdc.) varietas lokal Lembang di Kalimantan Selatan. Ziraa’ah. 43(2): 192-
197.
Kuswanto B, Waluyo RA, Pramantasari S, Canda. 2012. Koleksi dan Evaluasi Galur-
Galur Lokal Kacang Bogor (Vigna subterranea). Seminar Nasional Perhimpunan
Ilmu Pemuliaan Indonesia (PERIPI) tahun 2012. Institut Pertanian Bogor.
Muchtadi, D.2010. Kedelai Komponen untuk Kesehatan. Bandung (ID):
Alfabeta.
Muji I. et al. 2011. Isoflavone content and antioxidant properties of soybean seeds.
pp.16–20.
Nuraida, L., Suliantari, Andarwulan, N., Adawiyah D. R., Noviar, R. and Agustin
D. 2008. Evaluation of soybean varieties on production and quality of
tempe. In: Hardiansyah, Astawan, M., Kusumaningrum, Amelia, L.,
Briawan, D. and Aries, M. (Eds). Proceedings of Recent Developments On
Tempe: Technology, Standards, and Its Potency in Health and Nutrition
Improvement, p. 1. Bogor: Indonesian Tempe Forum, Yayasan Tempe
Indonesia, and Food and Nutrition Society of Indonesia.
Radiati A dan Sunarto. 2016. Analisis Sifat Fisik, Sifat Organoleptik, dan Kandungan
Gizi pada Produk Tempe dari Kacang Non-Kedelai. Jurnal Aplikasi Teknologi
Pangan. 5(1). 16-22.
Redjeki. 2009. Kacang Bogor. Jakarta (ID) : Gramedia Pustaka Utama.
Sitorus SR 2014. Perubahan alergenisitas protein kacang kedelai (Glycine max) dan
kacang bogor (Vigna subterranea) akibat pengolahan dengan panas. Jurnal
Aplikasi Teknologi Pangan. 2 (1) : 88-93.
Sukardi, Wignyanto, Purwaningsih I. 2008. Uji coba uji coba penggunaan
inokulum tempe dari kapang penggunaan inokulum tempe dari kapang
penggunaan inokulum tempe dari kapang rhizopus oryzae dengan substrat
tepung beras dan ubikayu pada unit produksi tempe sanan kodya malang.
Jurnal Teknologi Pertanian Vol. 9 No. 3 (Desember 2008) 207 – 215.
Tahir A. Anwar M. Mubeen H. Raza S. 2018. Evaluation of physicochemical and
nutritional contents in soybean fermented food tempeh by Rhizopus oligosporus.
Journal of Advance in Biology & Biotechnology 17(1):1-9.
Wilson S, Blaschek, de Meja E. 2005. Allergenic protein in soybean: processing and
reduction of P34 allergenicity. Nutr Rev, 63(2), pp.47–58.

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