Synthesis Of Silver Nanoparticles By Using Extracellular Metabolites Of Lactobacillus Delbrueckii subsp. bulgaricus

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FAOJLTY OF MATHEMATICS AND NATURAL SCIENCES,

BOGOR AGRICULTURAL UNIVERSITY

lPB

Internaoonal

Convention

Center

15 -1i'November 2011

Published

By

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ISBN : 978-979-95093-9-0

'PRO·CEEDI NGS

ISS 2013

International Seminar on Sciences 2013

"Perspectives on Innovative Sciences"

Bogar 15-17 November 2013

4P:B International Convention Center

Published by

Copyright© 2014

Faculty of Mathematics and Natural Sciences, Bogor Agricultural University

Proceedings of International Seminar on Sciences 2013 "Perspectives on Innovative Sciences"

Bog or 15-17 November 2013.

Published by: FMIPA-IPB, Jalan Meranti Kampus IPB Dramaga, Bogor 16680

Telp/Fax:

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http://finipa.ipb.ac.id

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395 pages

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Board of Editors

PROCEEDINGS

INTERNATIONAL SEMINAR ON SCIENCES 2013

Chief Editor:

Endar

H.

Nugrahani

Managing Editor:

lndahwati Nisa RC!chmania

Managing Team:

Wisnu Ananta Kusuma Ali Kusnanto

International Scientific Committee:

Manabu D. Yamanaka (Kobe University, Japan)

Kanaya (Nara Institute of Science and Technology, NAIST, Japan) Ken Tanaka (Toyama University, Japan)

Daniel Oosgood (Columbia University, USA)

Emmanuel Paradis (lnstitut de Recherche pour le Developpement, IRD, France) Rizaldi Boer (Begor Agricultural University, Indonesia)

Antonius Suwanto (Begor Agricultural University, Indonesia)

National Reviewer:

Kiagus Dahlan Tania June Sri Sugiarti Miftahudin Anja Meryandini

lmas Sukaesih Sitanggang

Farit Mochamad Afendi Paian Sianturi

Husin Alatas Heru Sukoco

,•.

t-FOREWORD

The International Seminar on Sciences 2013, which had the main theme "Perspectives on Innovative Sciences", was organized on November 15th -1th, 2013 by the Faculty of Mathematics and Natural Sciences, Bogor Agricultural University. This event aimed at sharing knowledge and expertise, as well as building network and collaborations among scientists from various institutions at national and international level.

Scientific presentations in this seminar consisted of a keynote speech, some invited speeches, and about 120 contributions of oral and poster presentations. Among the contributions, 66 full papers have been submitted and reviewed to be published in this proceeding. These papers were clustered in four groups according to our themes:

A. Sustainability and Science Based Agriculture

B. Science of Complexity

<:.Mathematics, Statistics and Computer Science

D. Biosciences and Bioresources

In this occasion, we would like to express our thanks and gratitude to our distinguished keynote and invited speakers: Minister of Science and Technology, Prof. Manabu D. Yamanaka (Kobe University, Ja pan), Prof. Kanaya (Nara Institute of Science and Technology, NAIST, Japan), Prof. Ken Tanaka (Toyama University, Japan), Emmanuel Paradis, PhD. (lnstitut de Recherche pour le Developpement, IRD, France), Prof. Dr .. Ir. Rizaldi Boer, MS (Bogor Agricultural University), and Prof. Dr. Ir. Antonius Suwanto, M.Sc. (Bogor Agricultural University).

We would like also to extend our thanks and appreciation to all participants and referees for the wonderful cooperation, the great coordination, and the fascinating efforts. Appreciation and special thanks are addressed to our colleagues and staffs who help in editing process. Finally, we acknowledge and express our thanks to all friends, colleagues, and staffs of the Faculty of

Mathematics and Natural Sciences IPB for their help and support.

Bogor, March

2014

The Organizing Committee

International Seminar on Sciences 2013

,' ' '·

Board of Editors

Foreword

Table of Content

Table of Content

Page

111

IV

v

A. Sustainability and Science Based Agriculture

2

3

4

5

6

7

8

9

Development of a natural rubber dryer Based on multi energy resources (biomass, solar and wind)

Didin Suwardin, Afrizal Vachlepi, Mili Pubaya, Sherly Hanifarianty

Characterization ofHDTMABr-modified Natural Zeolite and its Budi Riza Putra, Latifah K

Application in Cr(VI) Adsorption Darusman, Eti Rohaeti

Potency of Andrographis paniculata, Tinospora crispa, and Wulan Tri Wahyuni, Latifah K

Combjnation Extract as Darusman, Rona Jutama

.a-GJucosidase Inhibitor and Chromatographic Fingerprint Profile of the Extracts

Utilization of Frond Palrri Oil as Second Generation Bioethanol Deliana Dahnum, Dyah Styarini,

Production using Alkaline Pretreatment and Separated Hydrolysis Sudiyarmanto, Muryanto,

and Fermentation Method Haznan Abimanyu

Pretreatment of Grass Biomass with Biological Process for Efficient Hydrolysis

)

Alkaloid Compounds from Oil-Free Mahogany Seed (Swietenia macrophylla, King) and Hypoglycemia Effect of Mahogany Seed on The Rat (Rattus nov.ergicus)

Utilization OfVetiver Roots Waste Product as Strong, Low Density, and Eco Friendly Material Pot

Green Products from Wastewater of Tempe Industry

Saccharification of Oil Palm Empty Fruit Bunch After Alkaline Pretreatment Followed by Electron Beam Irradiation for Ethanol Production

Desy Kurniawati, Muhamad Natsir, Rahmi Febrialis and Prima Endang Susilowati

Sri Mursiti, Sabirin Matsjeh, Jumina, and Mustofa

Galuh Suprobo, Tatang

Gunawan, Cynthia Andriani, Rio Candra Islami

Susanti Pudji Hastuti, Yofi Brarnantya Adi, Bary Fratama, Samuel Arunglabi,

Dewi KAK Hastuti, and Santoso Sastrodiharjo

Muryanto, Eka Triwahyuni, Yanni Sudiyani

3

7

17

21

27

31

43

47

55

10 Isolation and Screening ofEndophytic Bacteria from Bark ofRaru Wasinton Simanjuntak, Heri 61

Plant (Tarrietia ribiginosa) and Their Potential for Bioetahnol Satria, and Nurul Utami

The Effect of Hypertension Herbs Formula to The Kidney Agus Triyono, Saryanto 67

Functions

12 The Use of Activated Carbon from Bintaro Fruit-Shell (Cerbera Anni Wulanawati, Kamella 71

manghas) as an Adsorbent to Increase Water Quality Gustina and Djeni Hendra

13 Analysis of Active Compounds from Mangosteen Rind (Garcinia Fachrurrazie, Harry Noviardi 77

mangostana L.) by Binding Affinity to The Androgen Receptor as

Anti-Prostate Cancer Drug Candidates

v v

14 Antioxidant Activity from Formula of Jati Belanda (Guazuma Syaefudin, Sulistiyani, Ed:c 81 (

ulmifolia Lamie), Jambu Biji (Psidium guajava Linn.), and Salam Djauhari Purwakusumah

'----'

(Eugenia polyantha Wight.) Leaves Extract

----·-15 Diversity of Bacterial Mercury Reductase Resistance (merA) from Prima Endang Susilowati, Sapto 87

Bombana Gold Mine Raharjo, Rachmawati Rusdin,

Muzuni

16 Brake Fem{Pteris vittata) as a Prospective Heavy Metal Mochamad Taufiq Ridwan, Rike 91

Accumulator: Utilization Potentials of Harvested Biomass and Tri Kumala Dewi and Agung

Heavy Metal Hasan Lukman

.

17 Protein Content Enhancement of Spirulina pl<:tensis by Irving Noor Arifin, lin 99

Phosphorus Limitation and Nitrogen Addition in Beef Cattle S'upartinah Noer and Asri Peni

Wastewater Medium Wulandari

18 Development immobilized enzyme of white-rot fungus for Ajeng Arum Sari and Samo 103

decolorization ofRBBR Tachibana

19 Simple and Rapid Screening Method for Early Identification of Sintho Wahyuning Ardie, Nurul 109

Salt Tolerant Foxtail Millet (Setaria italica L. Beauv) Khumaida, and Amin Nur

'

20 Synthesis of Silver Nanoparticles by Using Extracellular Suryani, Ridho Pratama, Dimas 113

Metabolites ofLactobacillus delbrueckii subsp. bulgaricus Andrianto /

B.

Science of Complexity

119

21 Regional Heat Capacity Changes due to Changes of Land Cover Winda Aryani, ldung Risdiyanto 121

Composition Using Landsat-5 TM Data

22 Microbial Cellulolytic Isolation and Identification from Durian Hapsoh, Gusmawartati dan 129

Leather Waste . Ujang AI Husnah

23 Predicting Water Surplus and Water Deficit in the Paddy Rice Johanis H . Panelewen, Johannes 135

Production Center in North Sulawesi Using the Water Balance E. X. Rogi and Wiske

Model _Rotinsulu3

24 Prediction of Dustfall Generation in Ambient Air over an AriefSilbdo Yuwono, Lia 143

Inceptisol Soil Area _Amaliah

" • \

25

Carboxymethylation ofMicrofibrillated Cellulose to Improve Fitri Adilla, Lisman 149

Thermal and Mechanical Properties of Poly lactic Acid Composites Suryanegara, Suminar S. Achmadi

26 Esterification ofMicrofibrillated Cellulose with Various Ajeng Mawangi, Lisman 155

Anhydrides to Improve Thermal and Mechanical Properties of Suryanegara, Suminar S.

Polylactic Acid Composite Achmadi

27 Thermal and Mechanical Properties Improvement of Poly lactic Resty Dwi Andinie, Lisman 161

Acid-Nanocellulose Composites by Acetylation Suryanegara, Suminar S.

Achmadi

C. Mathematics, Statistics and Computer Science

167

28 The comparison spatial distribution observed, estimatated using Rado Y endra, Ari Pani Desvina, 169

Neyman-Scott Rectangular Pulse Method (NSRP), and simulation Abdul Aziz Jemain for mean of one-hour rain and probability of24-hour rain

29 :Qptimal VARlnjectien -Based on Neural Network Current State Dimas Fajar Uman P, Ontoseno 175

:Estimator for 2-0kV Surabaya Electrical Distribution System Penangsang, Adi Soeprijanto

30 Fire-Fighting Robot Navigation System Using Wall Following Karlisa Priandana, Erwin M Y 181

Algorithm and Fuzzy Logic Chriswantoro, Mushthofa

31 Analysis and Solving of Outliers in Longitudinal Data Viarti Eminita, Indahwati, 187

Anang Kurnia

32 Implementation of Flowers and Ornamental Plants Landscape Meuthia Rachmaniah and 193

Information System using Cloud Computing Technology Iswarawati

33 Cluster Information of Non-sampled Area in Small Arca Rahma Anisa, Anang Kurnia, 199

Estimation with Non-normally Distributed Area Random Effects Indahwati

and Auxiliary Variables

34 Study of Overdispersion for Poisson and Zero-Inflated Poisson Lili Puspita Rahayu, Kusman 203

Regression on Some Characteristics of the Data Sadik, Indahwati

35 The Effect of Two-Way and Three-Way Interaction of Perceived Erui.y Kristiani, Ujang 209

Rewards on the Relationship Quality Sumarwan, Lilik Noor Yulianti

& Asep Saefuddin

36 Implementation oflnverse Kinematics for the Coordination Wulandari, Karlisa Priandana, 213

Control of Six Legged Robot Agus Buono

37 Detection of C Code Plagiarism by Using K-Means Ahmad Ridha, Abi Panca 219

Gumilang

38

Temporal Entity Tagging for Indonesian Documents Ahmad Ridha, Agus Simamora 223

39

Multidimensional Poverty Measurement Usir.g Counting Indah Soraya, Irwan Susanto, 229

4

5

·6

n

18

19

50

51

52

53

54

Minimizing

Linear

Optimization Model of Basic Reproduction

Number in a Fixed Number of Vaccination Coverage using Interior Point Method Approach

D. Chaerani, A. Anisah, N . Anggriani, Firdaniza

235

Expert System for Plant Growth using Hormones and Exogenous Yaasiinta Cariens, Karlina Nisa 241

· Factors based on Fuzzy Approach

The Effect of Divergent Branches on GPU-Based Parallel Program Hendra Rahmawan, Yudi Satria 247

Performance Gondokaryono

Ensemble of Extreme Estimates Based on Modified Aji Hamim Wigena, Anik 253

Champernowne and Generalized Pareto Distributions Djuraidah, Muhammad Hafid

Genetic Algorithms Application for Case Study of Septian Rahardiantoro, Bagus 259

Multi-Criteria Decision Analysis (MCDA) Sartono, Totong Martono

on the Data Contained Missing Value

An Implementation ofParallel AES Algorithm Aditya Erlangga, Endang 265

for Data Encryption with GPU Purnama Giri, Karlisa Priandana

Constructing Orthogonal Fractional Factorial Split-Plot Designs Bagus Sartono, Y enni Angraini, 269

by Selecting a Subdesign Dependently to Another Subdesign Indahwati

Spatial Clustering of Hotspots using DBSCAN and ST-DBSCAN Utsri Yustina Purwanto, Baba 275

Barus,and Hari Agung Adrianto

Gap between the ｾｯｷ･ｲ＠ and Upper Bounds for the Iteration Bib Paruhum Silalahi 281

Complexity oflnterior-Point Methods

Black Approximation To Determine Value Of Call Option On Jacob Stevy Seleky, Endar H. 287

Stock In Indonesian Stock Exchange Nugrahani, I Gusti Putu Pumaba

Analysis of Portfolio Optimization With and Without Shortselling Kaleem Saleem, Abdul Kohar 291

Basd on Diagonal Model: Evidence from Indonesian Stock Market Irwanto, Endar Hasafah Nugrahani

Community Network Framework as a Support of Successful Rina Trisminingsih, Christine 299

Agricultural Community Suryadi, Husni S. Sastramihardja

THE TRANSMISSION MODEL OF DENGUE FEVER Paian Sianturi, Ali Kusnanto, 305

DISEASE: A COMPUTER SIMULATION MODEL Fahren Bukhari

Improving the Independence of the Components of a Hari Wijayanto, Bagus Sartono, 311

Decomposition in Time Series Data Casia Nursyifa

Modeling and Empirical Mapping of Vehicular Traffic System: Endar H. Nugrahani, Hadi 322

Case Study of Jabodetabek Region Sumarno, Ali Kusnanto

. "

·.

55

56 57 58 59 60 61 62 63 64 65

D. Biosciences and Bioresources

A QuEChERS Based Method for The Determination of Pesticide Residues in Indonesian Green Coffee Beans Using Liquid Chromatography Tandem Mass Spectrometry

Design and Implementation of Roaster Control System Using Image Processing

Genetic Variation ofDGATllEael Gene of Holstein Friesian in National Dairy Cattle Stations

The Potency of Dahlia Tubers as Prebiotic for Functional Food

Harmoko, Rahmana Emran Kartasasmita, and Astika Tresnawati

Mohamad Agung Prawira Negara, Satryo Budi Utomo, Sumardi

Santiananda A. Asmarasari

Ainia Herminiati, Sri Pudjiraharti, Budi Setiawan

DNA identification using Markov Chain as feature extraction and Toto Haryanto, Habib Rijzaani,

Probabilistic Neural Network as classifier . Muhammad Luthfi Fajar

Multiple Sequence Alignment with Star Method in Graphical Processing Unit using CUDA

Abalone (Haliotis asinina) Wound Detection System Using Histogram and Morphology

Local Alignment of DNA Sequence Using Smith-Waterman Algorithm

Muhammad Adi Puspo Sujiwo, Wisnu Ananta Kusuma

Noer Fitria Putra Setyono, Aziz Kustiyo, Dwi Eny Djoko Setyono

Fariz Ashar Himawan, Wisnu Ananta Kusuma

Agronomic performance and yield potential of 18 com varieties in Anggi Nindita, Willy Bayuardi

Indonesia Suwamo, Surjono Hadi Sutjahjo,

Perdinan

Characteristic and Phisychochemical Properties of Sweet Potatoes Ai Mahmudatussa'adah (lpomoea batatas L)

Determination of Harvesting Time of Three Peanut Varieties Based on Heat Unit Accumulation

Heni Pumamawati, Yoga Setiawan Santoso, Yudiwanti Wahyu

66 Respon of Celery (Apium graveolens) Leaves Yield to Plant

Population and Seed Number Per Planting Hole

Karo, B, Marpaung, A. E., Tarigan, R., Barus, S. and Khaririyatun, N.

67 Bioethanol Production from Sugarcane Bagasses by Acid

Hydrolysis and Fermentation with Candida tropicalis

1,

Inda Setyawati , Laksmi Amba.rsari, Waras ｎｵｲ｣ｨｾｬｩｳＬ＠

,, Popi Asri Kumiatin, Puspa Julistia Puspita 323

325

·333 339 345 351 359 365 371 377 381 387 391 Ｏｾ＠

/

397

68 Toxic Compounds ofDichloromethane Extract ofMakasar Fruits Gustini Syahbirin, Rina Fazilatur 401

(Bruceajavanica) based on Artemia salina Leach Analysis Rahmi, Purwantiningsih Sugita

'

.,,

SYNTHESIS OF SILVER NANOPARTICLES BY USING

EXTRACELLULAR METABOLITES OF

Lactobacillus

delbrueckii

subsp.

bulgaricus

Suryani

1,

Ridho Pratama

2,

Dimas Andrianto

3

•.2.Jl Department of Biochemistry, Faculty of Mathematics and Natural Sciences Bogor Agricultural University, Indonesia

JI. Agatis, Kampus IPB Darmaga 16680, Bogor E-mail: ani3110@yahoo.com

Abstract

The silver nanopartic/es were used for many applications such as antifungal, anti bacterial, nanosensors, and food wrappers. Synthesis of silver nanopartic/es using microbes as well as metabolites have been developed with

consideration of the hazardous waste free environment. The aim of this research was to synthesize silver

nanopartic/es by ｵｳｩｾｧ＠ extracellular metabolites of L. delbrueckii subsp. bulgaricus. The synthesis silver

nanopartic/es was carried out by reacting the extracellular metabolites of L. delbrueckii subsp. bulgaricus with

silver nitrate (AgNOJ) solution. Reduction of Ag+ ions occurred by interaction of the extracellular enzyme that

produced by L. delbrueckii subsp. bulgaricus. The silver nanoparticles was detected by UV-Vis

spectrophotometer based on speci.fi.c absorption spectrum at the 400 nm wavelength. The result of Fourier Transform Infrared (FTJR) analysis showed that organic component played an important role in the silver nanoparticles formation. The enzyme was estimated as the organic component in the extracellular metabolites of

L. delbrueckii subsp. bulgaricus and had a role in the formation of silver nanoparticles. Furthermore, particles

size analysis conformed that average size of formed silver nanoparticles was 2. 7 nm and considered as silver

nanoparticles.

Keywords: AgN0_3, extracellular metabolites, Lactobacillus delbrueckii subsp. bulgaricus, synthesis of silver

nanoparticle

I.

INTRODUCTION

Silver nanoparticles have been used in various fields

such as antifungal (Vivek et al 2011), antibacterial

(Prasad et al 2011), nanosensor, pesticides, cleaners

for water and soil, food wrappers (Bouwmeester, 2007), also can reduce infections after surgery

(Kalishwaralal et al 2009).

. ｾｩｬｶ･ｲ＠ nanoparticles can be synthesized by

··· physically, chemically, and biologically (Kumar

.

ＺＭ ｾ ｾｑｰＩＮ＠

·

Synthesis of silver nanoparticles by

ｾＧ｜ ｲＮｾｙＮｓｩｾｬｬｹ＠ and chemically have disadvantages for its

·.-. _ility and aggregation (Kaliswaralal et al 2010) ,. poUute the environment. Synthesis of silver . . articles biologically is one of the alternative to \ke silver nanoparticles that more safe to the '· runent. Silver nanoparticles can be obtained :reduction of silver ions by microorganisms

ｾｳ＠ bacteria, yeast, or fungi (Tolaymat et al

, ｾ＠ - Synthesis of nanoparticles using

:raanisms are less expensive, non-toxic, high "vity, and easily adapted to the ambient

temperature and pressure (Reyes 2009). Therefore, silver nanoparticles have great potential to be developed in the future.

Synthesis of silver nanoparticles using the fungus

has also been reported by Sadowski et al (2008)

from Aspergillus niger as an agent biosynthesis.

Kumar & Mamidyala (2011) states that the

metabolites of Pseudomonas aeruginosa can be used

for extracellular biosynthesis of silver nanoparticles .

L. delbrueckii is a Gram positive, facultative

anaerobic, homofermentative, rod-shaped, not

sporulating, Synthesis of nanoparticles of this bacterium needs to be studied .

This aim of this research was to synthesis of silver '

nanoparticles by using extracellular metabolites of L .

delbrueckii subsp.bulgaricus. The result of this

research ｣｡ｮｾ ｰｲｯｶｩ､･＠ scientific information on the

biosynthesis of silver nanoparticles by reaction of

extracellular metabolites that safe for the

environment and can be applied in the field of medical and industries.

II. Methodology

a. Determination of Growth Curve

Lactobaci/lus delbrueckii subsp. bulgaricus was

refreshed on MRS medium and incubated at 42°C

for 16 hours at shaker incubator (120 rpm). The growth curve was determined by calculating the

optical density (OD) of Lactobacillus delbrueckii

subsp. bulgaricus after incubated at 0, 4, 8, 16, 32;

and 64 hours and measured the absorbance at 620 nm wavelength.

b. Synthesis of Silver Nanoparticles (Saravanan

et al 2011)

The bacteria were incubated at 42 ° C with agitation speed 150 rpm for 24 hours. Cells biomass and medium were separated by centrifugation (16000 g, 10 min). The 100 mL of supernatant culture were

mix:ed with 0.017 grams AgN03 and incubated in

dark chamber at room temperature. Mixture was allowed to stand for 30 minutes until the color changed. from clear to brown as an indicator of the

reaction between the supernatant and AgN03

c:

Analysis of Silver Nanoparticles With UV-Vis

· ' '" ··Spectrophotometer (Saravanan et al 2011)

Supernatant (reacted with AgN03) was taken 2 mL

for analysis by UV-Vis spectrophotometer and scanning at 200-800 nm wavelength. The formed peak will be analyzed for indication of silver · nanoparticles.

d. FfIR Analysis for Silver Nanoparticles (Kumar & Mamidyala, 2011)

Potassium bromideJKBr powder (200 mg) was made into tablets. The supernatant was dripped into potassium bromide tablet and then measured at wavelength 400-4000 cm·•. The results of frequency . spectrum measurement will further analyzed using correlation to determine the chemical bonds in organic compounds contained in the sample.

e. Analysis of of Silver nanoparticles Size by

Particles Size Analysis/PSA (Kim et al. 2006)

The refractive index and viscosity of solution that contained silver nanoparticles were analyzed first and then PSA. The average size of the silver nanoparticles were known by compared with size distribution of silver nanoparticles.

ID. RESULTS AND DISCUSSIONS

Growth curves of Lactobacillus delbrueckii subsp.

bulgaricus and AgN03 Treatment results

Biosynthesis of silver--nanoparti-cles"""'Were""""started

subsp. bulgaricus grown at 40-43°C as

optitn.ii·;,_

ｴ･ｭｰ･ｾｴｵｲ･Ｎ＠

The culture of L.

､･ｬ｢ｲｵ･ｾｫｩｩ＠

ｾｾ

ｫＲ＠

?ul?ancus were grown on ｾ＠ medium. ﾷｾｨｩｴＡ Ｈ ｾ＠

md1cated the changes of medium from clear: , ·, '

cloudy that caused by accumulation ofbiomas,s

｣［｜ｾ ＺＮ［＠

'. ;.

The aim of growth curves making is to detenni11e

optimum growth conditions of L. delbrueckii

Sl)bsp

bulgaricus. Observation was done for 64 hours ｡ｴｾ＠

2, 4, 8, 16, 32, 48, 64 hours after incubation (Figure

I). The L. delbrueckii subsp. bulgaricus

was

still

clear after 2 hours incubation as considered lag

phase i.e the phase of bacterial adaptation to

the

growth medium. Cell biomass increased only

slightly during this phase that caused slightly changed culture density. The log phase (exponential) started after 8 hours up to 16 hours incubation which indicated the optical density or turbidity value

increased and reached 1,754 (Figure 1). In this phase

L. delbrueckii subsp. bulgaricus have been able to

adapt to the medium and population growth increased exponentially (Black 2008). Furthermore,

the stages of growth deceleration occurs after 16

hours incubation time. In this phase, changes of

nutrition and environmental caused limitation of

bacterial growth.

2

l

1.8

-l

l

1.6

i

1.4

i

<!) !

ｾＱＮＲ＠

1

-e

1 Ｍｾ＠

o

I

ｾＰＮＸ＠

1

0.6 l

i

0.4 .,

I

0.2 ｾ＠

0

ｾ Ａ ｌＭ ＭｾＭ

ＭＭｾＭ

Ｍ ＭＭＮＭＭＭｾ

Ｎ＠

0 20 40

hours 60 80

Figure l. The turbidity growth curve of

Lactobacil/us delbrueckii subsp. bu/garicus

before after

Figure 2. Reaction with AgN03 before addition of

I •

.

...

· garvesting of L. delbrueckii subsp. bulgaricus for

. bjosynthesis of silver nanoparticles were carried out : ' after 24 hours incubation time when reached

. >statisioner phase. At the statisioner phase L.

· delbrueckii subsp. bulgaricus produced organic

compounds such as primary metabolites and secondary metabolites more than at the exponential pbaSe (Kunaepah, 2008). The next stage was

separation of cell biomass of L. delbrueckii subsp.

bulgaricus with growth medium containing

compounds the metabolites. Supernatant that

containing metabolites of L. de/brueckii subsp.

bulgaricus then added with AgN03 (0.017 g/100

mL).

There was color changes from clear

to

dark brown and became turbid after incubation for 30 minutes (Figure 2). Reaction of reduction directly observed by color changes of the solution (Kumar 2011). The color changes occurred due to the

reaction between organic compounds that

acclJJllulate in the bacterial growth medium

contained AgN0₃ that indicated bioreduction. Then,

UV - vis analysis was conducted to confirm whether

the compound formed silver nanoparticles.

Biosynthesis of Silver Nanoparticles by

Lactobaci/lus delbrueckii subsp. bulgaricus

UV-Vis Spectrophotometer Analysis Result The objectives for confirmation bioreduction of

AgN03 compound into silver nanoparticles was to

ensure that the color changes was biochemical reaction that indicates . the formation of silver nanoparticles. Indication of nanoparticles formation was known from peak absorption at 370-500 nm

wavelength (Solomon et al., 2007). In this research,

the formation of absorption peak was at 400 nm wavelength (Figure 3).

ｾ＠

QJ UI c

"'

t:

E

"' c

ft!

ｾ＠

ﾷｾﾷｾｍＭ ＭＭ

[image:13.599.33.295.29.790.2] [image:13.599.315.567.40.667.2]

·---;;:;;;----Wavelength

Figure 3. Absorption spectrum of silver

nanoparticles by L. delbrueckii subsp. bulgaricus

The absorption peak at 400 run wavelength indicated that there had been formation of silver nanoparticles

using extracellular metabolites L. delbrueckii subsp.

bu/garicus. According to Solomon et al. (2007),

absorption of silver formed at range 370-500 nm

wavelength. Analysis of UV Vis

spectrophotometer (Figure 3) provided information

the amount of nanoparticles formed. The higher

absorbance values can be assumed that the number of nanoparticles formed. Absorption peaks were very high that reached maximum absorbance value

was 4 that indicated formation of silver

nanoparticles in the solution .

Absorption spectrum was very important indicator to determine the formation of silver nanoparticles in the bioreduction of silver (Ag) ions. Visual observation of the color changes was not enough to prove whether or not the silver nanoparticles formation.

FfIR Analysis Result for the detection of Functional Groups in Formation of Silver N anoparticles

FTIR analysis results indicated that there were several strong absorption peaks, i.e at wavelengths

of 3432, 1642.1401, 1124 cm·1• The peak indicated

the bond contained in organic compounds in solution

of silver nanoparticles (Figure 4 ). Organic

compounds released by L. delbrueckii subsp.

bulgaricus had an important role in the bioreduction

of silver nitrate compounds. Spectrum with a

wavelength of 3432 cm·1 showed the content of

organic .compounds with the N-H bond of amines and amides compounds with moderate intensity.

Shaligram et al. (2009) stated that presence of amine

group indicated presence of proteins in biosynthesis of silver nanoparticles, because this compound was a peptide bond that linked amino acids to fonn proteins. FTIR analysis results indicated that there was an organic compound that played an important role during formation of silver nanoparticles

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Figure 4. Spectrum of FTIR in silver nano particles solution

These compounds ｾ｡､＠ role in reducing and wrap

silver nanoparticles to form stable and homogen sizes of nanoparticles. Protcin-suspected as one of organic component that had role in the formation of silver nanoparticles. Further analysis is needed to

determine specific protein or other organic

compounds that play a role in formation of silver nanoparticles. Reactions that occurred in formation of silver nanoparticles are oxidation and reduction that there was electron transfer between oxidation and reduction agent. The silver (Ag) ions was positively charged ions if reacted with organic compounds that produced by metabolic processes of bacteria will release electrons. This condition will

change silver (Ag} ions turn into Ag0• The enzyme

binded to the nitrate compounds from silver nitrate

(AgN03) and using nitrate as substrate. The enzyme

was nitrate reductase that played a role in the reduction of nitrate became nitrite.

The Size of Silver Nanoparticles by PSA

Analysis of the nanoparticles size was performed by

using PSA (Particle Size Analysis). PSA was

considered more accurate than Scanning Electron

Microscope (SEM). Transmission Electron

Microscope (TEM) and Scanning Force Microscope .(SFM), especially for samples in the order of nanometer and submicron which usually have a high agglomeration tendency (Lidiniyah 2011 ). PSA measurement result in the form of distribution can be used to determine the overall particle size. Measurements were performed at 25°C using refractive index l.3390 and viscosity samples were . 2.1700 cp. Value of refractive index and viscosity

will improve the accuracy of PSA measurement.

Based on the PSA analysis, it was known that average size of silver nanoparticles was 2.7 nm and PI (Polydispersity Index) was 0.351. PI value describes of the width of the size distribution of

particles. If PI value less than 0.3, it indicated that

particle size distribution is narrow and more homogeneous particle size, whereas PI values greater than 0.3, it indicated wide distribution and particle size tended more varied (Figure 5). The silver nanoparticles had PI value 0.351 that indicated

silver nanoparticles had homogen particles size. It

was proven that the metabolites of L. delbrueckii

subsp. bulgaricus had ability to synthesis of silver

nanoparticles when reacted with AgN03.

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---·-·-IV. CONCLUSIONS

Metabolites of Lactobacillus delbrueckii

SUbsp _

bulgaricus had ability to synthesize of

sil ·

nanoparticles when reacted with AgN0₃• . ; ; .

absorption peak of silver nanoparticles occurred ·

400 nm wavelength. The organic molecules

had

at

role in formation of silver nanoparticles by reducj '

AgN03 into silver nanoparticles. The size of

silvng

nano particles was 2. 7 nm. er

ACKNOWLEDGEMENT

We would like to thank for Directorate of General Higher Education Indonesia (DIKTn that provided this funding research (in 2012).

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