THE EFFECT OF DIFFERENT MEDIUM COMPOSITIONS ON PRODUCTION OF BACTERIOCIN OF LACTIC ACID BACTERIA ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT (Dendrocalamus asper) PICKLE UNDER DIFFERENT FERMENTATION CONDITIONS PENGARUH PERBEDAAN KOMPOSISI MEDIUM TERHADAP PRO

  

THE EFFECT OF DIFFERENT MEDIUM COMPOSITIONS ON

PRODUCTION OF BACTERIOCIN OF LACTIC ACID BACTERIA

  

ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT

(Dendrocalamus asper) PICKLE UNDER DIFFERENT

FERMENTATION CONDITIONS

PENGARUH PERBEDAAN KOMPOSISI MEDIUM TERHADAP

PRODUKSI BAKTERIOSIN DARI BAKTERI ASAM LAKTAT

YANG DIISOLASI DARI ACAR REBUNG BETUNG KUNING

  

(Dendrocalamus asper) DALAM KONDISI

FERMENTASI YANG BERBEDA

BACHELOR THESIS

  Submitted in partial fulfilment of the requirements for a Food Technology Bachelor's degree in Faculty of Agricultural Technology

  

By:

LORENTIA SANTOSO

12.70.0078

  

DEPARTMENT OF FOOD TECHNOLOGY

FACULTY OF AGRICULTURAL TECHNOLOGY

SOEGIJAPRANATA CATHOLIC UNIVERSITY

SEMARANG

2016

  

THE EFFECT OF DIFFERENT MEDIUM COMPOSITIONS ON

PRODUCTION OF BACTERIOCIN OF LACTIC ACID BACTERIA

  

ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT

(Dendrocalamus asper) PICKLE UNDER DIFFERENT

FERMENTATION CONDITIONS

PENGARUH PERBEDAAN KOMPOSISI MEDIUM TERHADAP

PRODUKSI BAKTERIOSIN DARI BAKTERI ASAM LAKTAT

YANG DIISOLASI DARI ACAR REBUNG BETUNG KUNING

  

(Dendrocalamus asper) DALAM KONDISI

FERMENTASI YANG BERBEDA

BACHELOR THESIS

  Submitted in partial fulfilment of the requirements for a Food Technology Bachelor's degree in Faculty of Agricultural Technology

  

By:

LORENTIA SANTOSO

12.70.0078

This thesis has been approved and defended in front of the examination

th

committees on 24 February 2016

nd

  Semarang, 2 March 2016 Faculty of Agricultural Technology Soegijapranata Catholic University

  Supervisor Dean, Dra. Laksmi Hartayanie, MP Dr. V. Kristina Ananingsih, ST, MSc Co-Supervisor Dr. Ir. Lindayani, MP

STATEMENT OF THESIS AUTHENTICITY

  I hereby declare that the thesis entitled

  “THE EFFECT OF DIFFERENT MEDIUM

COMPOSITIONS ON PRODUCTION OF BACTERIOCIN OF LACTIC ACID

BACTERIA ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT

(Dendrocalamus asper) PICKLE UNDER DIFFERENT FERMENTATION

CONDITIONS

  ” contains no work that ever proposed to acquire a bachelorship title in a

  University, and along to my knowledge, there is no work ever written or published by others, except the ones used as references in this thesis and mentioned in the list of references. If it is proven in the future that partially or whole thesis is the result of plagiarism, therefore I will be willing to be revoked with all the consequences in accordance with the law and regulations applied at Soegijapranata Catholic University and/or valid law and regulations.

  Semarang, March 2016 Lorentia Santoso

  12.70.0078

  

SUMMARY

Yellow Betung bamboo shoot is one of the bamboo shoot derived from an indigenous bamboo

species (Dendrocalamus asper) in Indonesia. However, bamboo shoot as one of foods has a

relatively short shelf life and easily damaged. One processing method to extend the shelf life of

bamboo shoot is fermentation process. Bamboo shoot that are fermented into pickled bamboo

shoot not only can have a longer shelf life, but also it can be one of the natural habitats for the

growth of lactic acid bacteria (LAB). One of the capabilities of LAB is to produce antimicrobial

compounds, such as bacteriocins. Bacteriocins of LAB were found to have antagonist activity

against pathogenic bacteria. However, studies on the influence of fermentation conditions and the

composition of the media to the bacteriocin production of LAB which isolated from Yellow

Betung bamboo shoot pickle is still limited. Therefore, the aim of this study was to determine the

antimicrobial activity of LAB bacteriocins isolated from Yellow Betung bamboo shoot pickle

under different fermentation conditions and the medium compositions. A total of eleven isolates

of LAB that have been previously isolated from pickled Yellow Betung bamboo shoot in four

different fermentation conditions, were tested for their ability to produce bacteriocins. Isolates

which isolated from the fermentation condition A (2.5% salt concentration at 15°C) were A20

and A43. While isolates obtained from the fermentation condition B (5% salt concentration at

15°C) were B1, B2, and B32. C18, C19, and C29 were isolated from the fermentation condition

C (2.5% salt concentration at 30°C). A total of three isolates were also isolated from the

fermentation condition D (5% salt concentration at 30°C), they were D11, D20, and D44. Isolate

selection for the production of bacteriocins was based on the ability to against pathogenic bacteria

S.aureus (FNCC 0047) and E.coli (FNCC 0091). Bacteriocin antimicrobial activity was tested by

removing the effect of organic acids produced by LAB by adjusting the pH of the cell-free

supernatant (bacteriocin) to 6.0. Testing the antimicrobial activity of bacteriocins was done by

agar-well diffusion method. The inhibitory activity was observed by measuring the clear zone and

_{2 -1}

calculate Activity Unit (AU in mm mL ) of bacteriocins. All LAB isolates were grown in media

without supplementation could not produce bacteriocins. Supplementation of carbon and nitrogen

sources can induce the production of bacteriocins by LAB. However, only six of LAB isolates

(Leuconostoc mesentroides ssp cremoris, Lactobacillus plantarum 1, Lactobacillus plantarum,

Lactobacillus fermentum 1, Lactobacillus pentosus) which isolated from bamboo shoot pickle

with fermentation condition C and D and also were supplemented with carbon and nitrogen source

showed inhibitory activity against S.aureus (FNCC 0047), E.coli (FNCC 0091), and

  Inhibitory activity magnitude of the neutralized cell-free L.monocytogenes (FNCC 0156). _{2 -1}

supernatant (bacteriocin) ranged from 113-4663 mm mL . The highest bacteriocin inhibitory

activity was found in bacteriocin D44 (produced by Lb.pentosus) which has been supplemented

with 2% glucose and 2% yeast extract. The inhibitory activity of bacteriocins D44 (Lb. pentosus)

_{2 -1 2}

against S.aureus (FNCC 0047) was equal to 4663 mm mL , E.coli (FNCC 0091) was 3109 mm

• _-1
_{2 -1}

mL , and L.monocytogenes (FNCC 0156) was 3179 mm mL . The results showed that the initial

fermentation conditions (differences in salinity and temperature of fermentation) could determine

different characteristics in each LAB produced mainly in terms of the ability to produce

bacteriocins as an antimicrobial compound. In this study, was also found that supplementation of

carbon and nitrogen sources were able to induce the production of bacteriocins which the result

depended on each LAB isolate.

  i

  

RINGKASAN

Rebung Betung Kuning merupakan salah satu jenis rebung yang berasal dari spesies bambu

(Dendrocalamus asper) asli Indonesia. Namun, rebung sebagai salah satu bahan pangan

memiliki umur simpan yang relatif singkat dan mudah mengalami kerusakan. Salah satu

pengolahan untuk memperpanjang umur simpan rebung adalah melalui proses fermentasi.

Rebung yang difermentasi menjadi acar rebung selain dapat memiliki umur simpan yang lebih

panjang, juga dapat menjadi salah satu habitat alami untuk pertumbuhan bakteri asam laktat

(BAL). Salah satu kemampuan BAL adalah menghasilkan beberapa senyawa antimikroba,

diantaranya adalah bakteriosin. Bakteriosin dari BAL ditemukan memiliki aktivitas antagonis

terhadap bakteri patogen. Namun, studi tentang pengaruh kondisi fermentasi dan komposisi

media terhadap produksi bakteriosin BAL yang diisolasi dari acar rebung Betung Kuning masih

terbatas. Oleh karena itu, tujuan penelitian adalah untuk mengetahui aktivitas penghambatan

bakteriosin dari BAL yang diisolasi dari acar rebung Betung Kuning dalam kondisi fermentasi

dan komposisi media yang berbeda. Sebanyak sebelas isolat BAL yang telah diisolasi sebelumnya

dari acar rebung Betung Kuning dalam empat kondisi fermentasi yang berbeda, diuji

kemampuannya dalam menghasilkan bakteriosin. Isolat yang diambil dari hasil fermentasi kode

A (2,5% kadar garam pada suhu 15°C) adalah A20 dan A43. Sedangkan isolat yang diperoleh

dari hasil fermentasi kode B (5% kadar garam pada suhu 15°C) adalah B1, B2, dan B32. Dari

hasil fermentasi kode C (2,5% kadar garam pada suhu 30°C) diambil isolat C18, C19, dan C29.

Sebanyak tiga isolat juga diambil dari hasil fermentasi kode D (5% kadar garam pada suhu

30°C), yaitu D11, D20, dan D44. Pemilihan isolat untuk produksi bakteriosin didasarkan pada

kemampuannya dalam menghambat bakteri patogen S.aureus (FNCC 0047) dan E.coli (FNCC

0091). Daya hambat bakteriosin diuji dengan cara menghilangkan efek asam organik yang

dihasilkan oleh BAL dengan menyesuaikan pH supernatan bebas sel (bakteriosin) menjadi 6,0.

Pengujian daya antimikroba dari bakteriosin dilakukan dengan metode difusi sumuran agar.

Aktivitas penghambatan diamati dengan mengukur zona bening dan menghitung Activity Unit

_{2 -1}

(AU dalam mm mL ) dari bakteriosin. Semua isolat BAL yang ditumbuhkan pada media tanpa

suplementasi tidak dapat menghasilkan bakteriosin. Suplementasi sumber karbon dan nitrogen

dapat menginduksi produksi bakteriosin oleh BAL. Namun, hanya enam isolat BAL (Leuconostoc

mesentroides ssp cremoris, Lactobacillus plantarum 1, Lactobacillus plantarum, Lactobacillus

fermentum 1, Lactobacillus pentosus) yang diperoleh dari acar rebung kondisi fermentasi C dan

D yang telah disuplementasi dengan sumber karbon dan nitrogen yang mampu menunjukkan

aktivitas antimikroba dalam menghambat indikator patogen (S.aureus (FNCC 0047), E.coli

(FNCC 0091), dan L.monocytogenes (FNCC 0156)). Besarnya aktivitas penghambatan

( AU/Activity Unit) dari supernatan yang telah dinetralkan (bakteriosin) berkisar antara 113-

_{2 -1}

4663 mm mL . Aktivitas penghambatan yang tertinggi ditemukan pada bakteriosin yang

dihasilkan oleh isolat D44 (Lb. pentosus) yang telah disuplementasi dengan 2% glukosa dan 2%

ekstrak yeast. Aktivitas penghambatan bakteriosin D44 (Lb. pentosus) dalam menghambat

_{2 -1 2 -1}

S.aureus (FNCC 0047) sebesar 4663 mm mL , E.coli (FNCC 0091) sebesar 3109 mm mL , dan

_{2 -1}

L.monocytogenes (FNCC 0156) sebesar 3179 mm mL . Hasil penelitian menunjukkan bahwa

kondisi fermentasi awal (perbedaan kadar garam dan suhu fermentasi) dapat menentukan

karakteristik yang berbeda pada setiap BAL yang dihasilkan terutama dalam hal kemampuan

menghasilkan senyawa antimikroba bakteriosin. Diketahui juga bahwa suplementasi sumber

karbon dan nitrogen mampu menginduksi produksi bakteriosin yang hasilnya tergantung pada

masing-masing isolat BAL.

  

ACKNOWLEDGMENT

  Firstly, I dedicated my greatest gratitude to Jesus Christ and The Virgin Mary, who have given their affection and blessing to the author for taking the time to complete this thesis entitled

  “THE EFFECT OF DIFFERENT MEDIUM COMPOSITIONS ON THE PRODUCTION BACTERIOCIN OF LACTIC ACID BACTERIA ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT (Dendrocalamus asper) PICKLE UNDER DIFFERENT FERMENTATION CONDITIONS”. It has been written to fulfill the graduation requirements of the Food Technology Program at the Faculty of Agricultural Technology. I was engaged in researching and writing this thesis from September 2015 to February 2016. This thesis is a part of the second year of Penelitian Unggulan Perguruan Tinggi (PUPT) 2015/2016 entitled

  “Efek Probiotik dan Mikrostatik dari Bakteri Asam Laktat yang Berper an dalam Fermentasi Acar Rebung”.

  The author really gives regards and thanks for people who has given guidance and help; they are:

  1. Dr. Victoria Kristina, ST, MSc, as the Dean of Faculty of Agricultural Technology Soegijapranata Catholic University Semarang, for giving the chance to finish this final research.

  2. Dra. Laksmi Hartayanie, MP as my supervisor and Dr. Ir. Lindayani, MP as my co- supervisor. I also wish to thank for their excellent guidance and support during this process. Thank you for always available and willing to answer my queries, without their cooperation I would not have been able to conduct this research.

  3. Elizabeth Ivone Fernandez, SSi, MSc as thesis coordinator for arranging the administration and schedule during this process.

  4. All lecturers in Food Technology Department for being a great teacher for giving the joyful of learning.

  5. The laboratory assistants for all their help and motivation throughout the unforgettable laboratory life in Food Technology Department.

  6. The administration staff for providing great service and information during the study activities. iii

  7. My parents (Go Roy Santoso and Liliawati) and my beloved brothers (Yafet Haere, Danny Setiawan, and Surya Santoso) for always being number one supporter and saying my name in every prayer. If I ever lost interest, you kept me motivated.

  8. Ivana Aprilia Pratiwi and Veronica Dian Sari who had been perfect sisters during campus life.

  9. Anastasya Gumelar, Amelia Juwana, Melita Mulyani, Vivian, Michaela Jessica, Matius Inda, Lukas Terry, Anton Septian, Hendra Aditya, Michael Gurdamulya, Hengky Kurniawan, Michael Yefta, Asien, Toni Hartanto, Arnulf Budi, LIMUN Team, Student Organization Team, GLORY Team, Assistant Laboratory Team, AU Internship Team, all of student in Food Technology who had been a part of the author’s experiences during campus life.

10. All people who has directly and indirectly helped during the experiment until finishing the thesis report.

  Finally, the author realizes that there were unintended errors in writing this report. The author really allows all readers to give suggestions to improve its content in order to be made as one of the good examples for the next thesis. Big hope from the author that this report can be useful for others.

  Semarang, March 2016 Author,

  Lorentia Santoso iv

  

LIST OF CONTENTS

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LIST OF TABLES

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LIST OF FIGURES

  

LIST OF APPENDICES

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