LACTIC ACID BACTERIA

ISOLATED FROM
FERMENTED FOODS
MADE OF INDONESIAN
LOCAL SOURCES:
POTENTIAL AND
APPLICATION
Amelia Juwana, Cynthia Christine, Tesyara Danesh, Cindy Kusuma, Lindayani, Laksmi Hartayanie
Soegijapranata Catholic University
Semarang

The 2nd ICSAF, Semarang, Indonesia
BACKGROUND
still become the most beneficial
microorganisms due to their probiotic
potential

have inhibitory activities
such as antimicrobes
WHY LAB?
potential to be used as preservatives
The 2nd ICSAF, Semarang, Indonesia
BACKGROUND
LAB LAB have complex nutritional requirements because of
their limited biosynthetic capabilities.

Indonesia has plenty local sources that contain
protein and sugar units, which should be suitable
& environments for LAB.
Indonesian fermented foods have been consumed
INDONESIAN for centuries, but there are little investigations
LOCAL SOURCES have been conducted to assess the diversity of
LAB in Indonesian fermented foods.
The 2nd ICSAF, Semarang, Indonesia
BACKGROUND

This study has been conducted to evaluate probiotic
potential of LAB which already screened from mandai
(fermented cempedak (Artocarpus champeden Spreng)
dami), sayur asin (fermented sawi pahit (Brassica juncea
(L.)), betung bamboo shoot (Dendrocalamus asper) pickle,
fermented Lindak cocoa (Theobroma cacao L.) seed and
viability of LAB isolates in food products during storage.

The 2nd ICSAF, Semarang, Indonesia
MATERIAL & METHOD
SOURCES of LAB

SAWI PAHIT

The 2nd ICSAF, Semarang, Indonesia
CEMPEDAK BETUNG BAMBOO SHOOT LINDAK COCOA SEED

MANDAI SAYUR ASIN BETUNG BAMBOO SHOOT FERMENTED LINDAK
PICKLE
MATERIAL & METHOD
Screening Test
Incubated for  Gram Staining
30-37º C,  Catalase Test
16-48 hours  Motility Test
 Growth at Different Temperature, NaCl and pH

Transferred
5 isolates
to test tubes Screening Probiotic and Antimicrobial
Detection
 Acid Tolerance
 Bile Salt Tolerance
 Antimicrobial Activity Detection

Identification of LAB
 Molecular Identification
 API Reading
The 2nd ICSAF, Semarang, Indonesia
MATERIAL & METHOD
Molecular Identification
Mandai ON GOING
DNA Fingerprinting

Sayur API Reading Lactobacillus plantarum 1 Purple Sweet
Asin (Sig. 96.6%) Potato Concentrate
(PSPC)
IDENTIFICATION Betung
Bamboo API Reading Lactobacillus plantarum
OF LAB Minced Beef (MB)
Shoot (Sig. 97.7%)
Pickle

Fermented API Reading
Lactobacillus plantarum Pineapple Juice
Lindak
(Sig. 99.9%) (PJ)
Cocoa
Seed
The 2nd ICSAF, Semarang, Indonesia
MATERIAL & METHOD
Purple Sweet PSPC+ LAB
Sayur Asin
Lactobacillus plantarum 1 Potato Concentrate
(PSPC) PSPC + LAB + Sodium Benzoate

Betung Bamboo Shoot Pickle MB + LAB
Lactobacillus plantarum Minced Beef (MB)
MB + LAB + Nitrite

Fermented Lindak Cocoa Seed Pineapple Juice PJ+ LAB
Lactobacillus plantarum (PJ)
PJ + LAB + Sodium Benzoate

Qualitative Viability Test
(day 1-4)
The 2nd ICSAF, Semarang, Indonesia
API READING MICROTUBES
RESULT AND DISCUSSION
Acid Tolerance
Acid Tolerance Acid Tolerance
Source Isolate pH 3 pH 7 Source Isolate pH 3 pH 7
0h 1.5h 3h 0h 1.5h 3h 0h 1.5h 3h 0h 1.5h 3h
A2 + + + + + + Betun 1.1.S + + + + + +
A14 + + + + + + g 2.1.S + + + + + +
Mand B30 + + + + + + Bambo 4.1.S + + + + + +
ai o
E52 + + + + + + 5.1.S + + + + + +
Shoot
F71 + + + + + + 6.2.S + + + + + +
Pickle
43J + + + + + + 23 + + + + + +
22J + + + + + + Ferme
nted 32 + + + + + +
Sayur 52J + + + + + +
Asin Lindak 33 + + + + + +
42J + + + + + + Cocoa 35 + + + + + +
71J + + + + + + Seed 47 + + + + + +
The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Bile Salt Tolerance
Bile Salt Tolerance Bile Salt Tolerance
Source Isolate 0.3% Source Isolate 0.3%
0h 2h 4h 0h 2h 4h
A2 + + + 1.1.S + + +
A14 + + + Betung 2.1.S + + +
B30 + + + Bamboo 4.1.S + + +
Mandai
Shoot
E52 + + + Pickle 5.1.S + + +
F71 + + + 6.2.S + + +
43J + + + 23 + + +
Ferment
22J + + + 32 + + +
ed
Sayur 52J + + + 33 + + +
Lindak
Asin
42J + + + Cocoa 35 + + +
71J + + + Seed 47 + + +
The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Detection of Antimicrobial Activity
Inhibition Zone (ZOI) (mm)
Source Isolate Escherichia coli Staphylococcus aureus Listeria monocytogenes Salmonella typhimurium

A2 12.03  13.37 2.73
A14 8.87  9.40 5.03
Manda B30 9.70  10.43 9.00
i
E52 7.37  10.33 6.03
F71 10.73  13.73 6.40
43J 6.2-9.2 10.8-11.6  
22J 10.0-10.9 7.7-14.7  
Sayur 52J 9.2-10.7 4.8-10.4  
Asin
42J 6.2-8.7 9.9-13.4  
71J 4.6-6.9 9.8-14.5  
The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Detection of Antimicrobial Activity
Inhibition Zone (ZOI) (mm)
Source Isolate Escherichia coli Staphylococcus aureus Listeria monocytogenes Salmonella typhimurium

1.1.S 10.6-11.8 11.6-12.9  
Betung 2.1.S 9.5-10.3 10.9-12.7  
Bamboo 4.1.S 7.7-8.8 8.5-14.7  
Shoot
Pickle 5.1.S 9.2-12.5 11.0-11.8  
6.2.S 8.4-11.9 10.8-14.0  
23 8,60-8,80 9,75-11,60  
Fermented 32 7,60-12,20 7,50-13,00  
LIndak 33 6,70-12,25 11,00-16,00  
Cocoa
Seed 35 8,60-11,35 6,00-6,50  
47 9,90-12,90 7,00-10,40  
The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test
Storage at 4° C (Day)
Source Product Isolate Treatments 1 2 3 4
Sayur PSPC 43J PSPC + 43J + + + +
Asin 22J PSPC + 22J + + + +
52J PSPC + 52J + + + +
42J PSPC + 42J + + + +
71J PSPC + 71J + + + +
43J PSPC + Sodium Benzoate + 43J + + + +

22J PSPC + Sodium Benzoate + 43J + + + +

52J PSPC + Sodium Benzoate + 52J + + + +

42J PSPC + Sodium Benzoate + 42J + + + +

71J PSPC + Sodium Benzoate + 71J + + + +

The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test

The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test
Storage at 4° C (Day)
Source Product Isolate Treatments
1 2 3 4
Betung MB 1.1.S MB + 1.1.S + + + +
Bamboo 2.1.S MB + 2.1.S + + + +
Shoot
Pickle 4.1.S MB + 4.1.S + + + +
5.1.S MB + 5.1.S + + + +
6.2.S MB + 6.2.S + + + +
1.1.S MB + Nitrite + 1.1.S + + + +
2.1.S MB + Nitrite + 2.1.S + + + +
4.1.S MB + Nitrite + 4.1.S + + + +
5.1.S MB + Nitrite + 5.1.S + + + +
6.2.S MB + Nitrite + 6.2.S + + + +

The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test

The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test
Storage at 4° C (Day)
Source Product Isolate Treatments
1 2 3 4
Fermented PJ 23 PJ + 23 + + + +
Lindak 32 PJ + 32 + + + +
Cocoa
Seed 33 PJ + 33 + + + +
35 PJ + 35 + + + +
47 PJ + 47 + + + +
23 PJ + Sodium Benzoate + 23 + + + +
32 PJ + Sodium Benzoate + 32 + + + +
33 PJ + Sodium Benzoate + 33 + + + +
35 PJ + Sodium Benzoate + 35 + + + +
47 PJ + Sodium Benzoate + 47 + + + +

The 2nd ICSAF, Semarang, Indonesia
RESULT AND DISCUSSION
Qualitative Viability Test

The 2nd ICSAF, Semarang, Indonesia
CONCLUSION
All LAB isolated from mandai (fermented cempedak (Artocarpus champeden
Spreng) dami), sayur asin (fermented sawi pahit (Brassica juncea (L.)), betung
bamboo shoot (Dendrocalamus asper) pickle, fermented Lindak cocoa
(Theobroma cacao L.) seed have probiotic potential based on screening
probiotic results and were able to inhibit the growth of indicator pathogenic
bacteria based on detection of antimicrobial activity results.

Qualitative viability test showed that all LAB isolated from betung bamboo shoot
(Dendrocalamus asper) pickle, sayur asin (from sawi pahit (Brassica juncea
(L.)), fermented Lindak cocoa seed (Theobroma cacao L.) which applied to
minced beef, purple sweet potato concentrate, and pineapple juice were able to
grow under storage at 4° C for 4 days with and without addition of synthetic
preservatives.
The 2nd ICSAF, Semarang, Indonesia
REFFERENCES
 Iñiguez-Palomares C., Pérez-Morales R., Acedo-Félix E. (2007). Evaluation of Probiotic Properties in
Lactobacillus Isolated from Small Intestine of Piglets. Revista Latino Americana de Microbiologia 49(3-
4):46-54.

 Ishola, R.O.and B.C.A-Tayo. (2012). Screening of Lactic Acid Bacteria Isolated from Fermented Food for
Bio-molecules Production. Assumption University Journal of Technnology 15(4):205-217.

 Rahayu, E.S. (2003). Lactic Acid Bacteria in Fermented Foods of Indonesian Origin. Journal of Agritech
Gadjah Mada University.

 Tamang, J.P., S. Dewan, B. Tamang, A. Rai, U. Schillinger, W.H. Holzapfel. (2007). Lactic Acid Bacteria
in Hamei and Marcha of North East India. Indian Journal of Microbiology 47: 119-125.

 Yavuzdurmaz, H. (2007). Isolation, Characterization, Determination Of Probiotic Properties Of Lactic
Acid Bacteria From Human Milk. Thesis. Izmir Institute of Technology. (unpublished)

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