Environmental Protection ancl Management - FTSL-ITB

Proceecling

Environmental Protection ancl Management

The l't International _Conference on Sustainable Infrastructure

ancl Built Environment in Developing _Countries

SABUGA ITB, Banclung - Indonesia

Znd

- 3'd November ZAO?

i

Publishect by

Faculty of civil ancl Environmental Engineering Institut Tel<nologi Bandung - ^Indonesia

@AUN/'EED-N", jffi _ffi"

5l B(_-200

9IBE 2009 published eight (8) volumes

of

proceeding a,s

following

^:

Volume

A

: Structr4re and

Material

Volume

B

Volume C Volume

D

Volume

E

Volume

F

Volume G :

Transportation System and Engineering Water Engineering and Management Waste Engineering and Management Ocean Engineering

C ons tructi o n

M

anagement

Ge o t e chnic al E ngineering

volume

H

: Environmental Protection and Managernent

rrt-rutuf;n$#u,', susrainabre,"*,J,X'oillt#5'l""iiffil::lfrfiliiilu",oo,,.E _counrries i^!r'+tii$ff;*i#j: _November 2d-3. 2009, Bandung,West Java ^_ tnd-Jnesia

PREFACE

The lst

International Conference

on

Sustainable Infrastructure and

Built

Environment in Developing Countries (SIBE) 2009 is'aimed to provide a forum to discuss and disseminate recent advance in scientific research, technology, and management upptourf, to obtain better environment quality.

Infrastructure that provides the basic need

of

a society and sustainable infrastructure system are essential for the survival, health and well-being of a society. In developing countries,

civil

and environmental engineers are at the epicenter

in

seeking means to enhance the quality

of

human

life

through modernization

of

infrastructure as evidenced

by

provision

of

shelters, water, and transPofr, amongst others. The current rate

of

wbanization and industrialization raises a number

of

environmental issues, often resulting

in

environmental mismanagement, especially

in

developing countries. The problems

ur" fr.ther

aggravated

by

environrnental degradation such as

soil

erosion, depletion

of

water resources, etc. I;n order

to

meet these multifaceted challenges' proper planning followed

by

implementation and verification must be exercised, via an integrated, multi disciplinary and holistic approach.

The conference

will

provide an opportunity for professionals and researchers to learn, share

and

exchange about

the

latest development

and

research

in civil and

environmental engineering.

The

scope

of the

conference covers

all

aspect

of civil

and environmental engineering practices.

Participants of the conference include researchers, academic staffs, sfudents, industries, public and local govemments. The keynote presentations during the conference are as follows:

Keynote speakers:

.

Indonesian GovernmentRepresentative Minister of Public Works, Indonesia

o

Dr. Puti Farida

Marzuki

Dean of ^the Faculty of Civil and Environmental Engineering, Institut Telmologi Bandung, Indonesia

o Dr.

Tony

Liu

National Taiwan University, Taiwan

r

Prof. Shunji Kanie

Hokkaido University, Japan

.

Prof. Syunsuke Ikeda

Tolryo Institute of Technologt

(AwsEED-Net),

_Japan.

Invited speakers:

o Dr.

Setiawan Wangsaatmaja

Environmental Protection Agency of West Java Province, Indonesia

. Dr.

Edwan Kardena

Faculty of Civil and Environmental Engineering, Institut Tetcnologi Bandung, Indonesia

.

Prof. Harianto Rahardjo, ^ph.D.

School of Civil and Environmental Engineering, Nanyang Technological (Jniversity, Singapore

.

Prof. Ikuo Towhata

School of Engineering, University of ^Totryo, Japan

.

Prof.

Dr.

Seiichi Kagaya

School of Engineering, Hokkaido (Jniversity, Japan

.

Prof. Jun Sasaki

school of Engineering, Yokohama National (Jniversity, Japan

.

Prof. Carl

Martin

Larsen

Norwegian university of science and Technology, Norway

The 1* InlernaUonal Conference on

Sustainable InfrasUuture and Built Environment ln Developing CoJntries November 2'n-3d 2009, Bandung,West Java - Indonesia

Dr.

George W. Annandale, D.Ing., P.E.

Golder Associates Inc., USA

David'Woodward, D.Phil.

University of Ulster, ^United Kingdom Dr. Delia

Dimitriu

Centrefor

Air

Transport and the,Environment (CATE), Mancltester Metropolitan University, United Kingdom

Prof. Tsunemi Watanabe

Department of Infrasttucture Engineering, Kochi (Jniversity of Technolog,t, Japan Prof. Shyh-Jiann Hwang

Department of Civil Engineering, National Taiwan University, ^Taiwan Prof. Kuo-Chun Chang

Department of Civil Engineering, National Taiwan University, Taiwan Prof. Suprihanto Notodarmojo

Faculty of Civil'and Environmental Engineering, Institut Tehtologi Bandung, Indonesia Masyhur Irsyam, Ph.D

Faculty of Civil and Environmental Engineering, Institut Teknologi Bandung, Indonesia Prof. Nakasaki

Kiyohiko

Tol+yo Institute of Technologt, Japan.

The objectives of this conference are:

1. ^To

^provide

^{a platform} for

exchange

of

ideas, information and experiences among academics, researchers, consultants, engineers, manufacturers and post graduate scholars in civil and environmental engineering.

2.

To discuss and evaluate the latest approaches, innovative technologies, policies and new

directions in

infrastructure development,

pollution prevention and

eco-friendly technologies adapted to developing countries.

3.

To promote cooperation and networking amongst practitioners and researchers involved in addressing infrastructure and built environment issues.

The oral and poster presentations are subdivided into 8 major sections, as following:

A.

Structure and material

B.

Transportation system and engineering

C.

Water engineering and management

D.

Waste engineering and management

E.

^Ocean engineering

F.

Construction management

G.

Geotechnicalengineering

H.

Environmental protection and management.

There are 174 contibutors in oral presentation and 36 contributors for poster presentation.

Finaly, the Organizing Committee wishes that this conference

is

able to provide beneficial scientific information to the participants and other concerned readers.

Bandung, November 2009 Organizing Committee

rts{,-e.qsp-*,

,'*gS#fs

susrainabre,",1..,o1"r1.:t#;'ffifi;icr;1ff:illi novemuert^riJtobs,Cil;;;;w;;i1.".-rndonesiail".rop,,,g countries

LIST OF COMMITTEE

STEERTNG

COMMITTEE

Chair

Co-Chair

Dr. Puti F. Marzuki

(Dean of FCEE ITB, Indonesia) Prof. Indratmo Soekarno Prof. K.C Chang

Prof. Bambang Sugeng Dr. Biemo W. Soemardi Prof. Arwin Sabar Dr. Ricky L. Tawekal

Prof. Takashi Mikami Dr. Iswandi Lnran Dr. Masyhur Irsyam

Prof. Suprihanto Notodarmodjo Prof. Enri Damanhuri

ORGANIZING COMMITTEE

Chair Co-Chair

General Secretary

Prof. Dr. h. Enri Damanhuri Ir. Muhamad Abduh,MT, Ph.D.

Dn Kania Dewi, ST, MT.

Dr. Hendra Achiari,ST, MT.

Ir. Rildova, MT, Ph.D.

Dr. Russ Bona Frazila, ST, MT.

Ir. Erza Rismantojo, Ph.D.

Ir. Ima Fatima, M.Eng.

Dr. Ir. Tri Padmi Dr. Sukandar, S.Si, MT.

Dr. Endra Susila, ST,

MT

Suphia Rahmawati, ST, MT.

Ivindra Pane,ST,MSE, Ph.D.

Ms. Iin Varida Mr. Ibet Setiawan

Mr. Rahmat Danu Andika

I Made Wahyu Widyarsana,ST,MT.

Dhemi Harlan,ST,MT,M Sc.,Ph.D.

Dr. Mochammad Chaerul Dr.

k.

Dwina Roosmini, MS.

Dr.

Heto

Dwi Ariesyady,ST,MT.

Dr. Nita Yuanita, ST, MT.

Qomanrdin Helmy, ^ST, MT.

Hasbullah Nawir, ST, MT, Ph.D.

Dr.

k.

Sony Sulaksono, MT.

Ms. Rahima Hanifa Ms. Neneng Eliana Mr. Boby Tri Andono

SCMNTIFIC COMMITTEE

Chair Co-Chair

Dr. Krishna S. Pribadi Dr. Saptahari Soegiri Prof. Naoyuki Funamizu Prof. Rizal Z. Tarnin Prof. Amrinsyah Nasution Prof. Wisjnuprapto Dr. Wayan Sengara

Dr. Syahril B. Kusuma

Prof. Shyh-Jiann Hwang Prof. Hattg Tuah

Prof. O&ar Z. Tarnin Prof. Adang Surahman Prof. Juli Soemirat Dr. R. Driejana

5re{-r&,qos;.." sustainabre

rnfra',Iff-[l!!fii*Hlfrfilii

su"rop,ng countries

"

j',Yq$ftilf;;il*'

_November

i'-j;

^iooq rji"J;;,i,il;;iil, :iil"jn"r,,

CONTENT

PREF'ACE

LIST

OF

COMMITTEB

PLENARY

LECT{IRE

Djoko Kirmanto

tr(eynote Speech

from

the

Minister

_{of Public Works} Republic of

Indonesia

1

Puti Farida Marzuki

Toward Sustainable

Infrastructure

_and

Built

Environment: Striving

for

Relevance

in Civil

and Environmental Engineering Education and

Research

₅

Shunji Kanie

Sustainable Development in vulnerable Environments: For construction _and

Engineering in permafrost

Regions

₁₆

Syunsuke lkeda

Transport Processes of Sediment /I.{utrients in Watershed and Application to Asian and pacific _Countries

'r<

Tony c. Liu, Jenn-chuan chern, and Kuo-chun chang

concrete Technorogy

for

sustainable

Infrastructure

₃₅

SPECIAL

LECTURB

ORAL SESSION PAPERS

Abdullah Al-Mamun, Maan Fahmi R. At-Khatib, Aishah Jamaluddin Ahmad, Mohammed A.

Al Sa'adi, Md, Zahangir AIam, Suleyman Aremu Muyibi, Faris Ismail, Azni Idris Adsorption of Chromium from Water by Carbon Nanotubes Grown on Powdered Activated Carbon

Amitava

Bandopadhyay

⁴²

Recent rnnovations in Managing _and

utilization

_{of solid Wastes}

from

Steel Sector 49 Bui Xuan Thanh, Chettiyapan Visvanathan

Performance of Aerobic _Granules Coupling

With

Membrane

Bioreactor

_5s

Dima Jawad, Mirielle Manoukian

Developing

LCCA

Model

for

Solid Waste Management Facility: What is

Important?

^r

-

Effendi, A.J., _Q. Helmy., ^{p.^S.} Hoesni, T. Tedjakusuma 62

The Application _of Bioemulsifier Produced by Azotobacter vinelandii

in

Bioremediation Oil-Contaminated

SoiI

₆₉

Fadjari Lucia Nugroho, Setiati, Roby

Biosorption of Remazol Golden Yellow 6 Dye by Mixed

culture

of Live

Fungi

7s Florinda Bacani, Gonzalo chua Jr., Elvin Dexter Lee, Cedrik Niget perfecto

Test and

Monitoring

_{of Lead} Accumulation _Using Hydrophyte _Hibiscus

Cannabinus

L'PIant

as Metal Accumulators

for

Potential

phytoremediation

_g2

I UI

1V

The lsr International Conference on

'

r

t-{6f*$&

^{s u sta ina b'|:,rff} _:lfpjs ffsr: H:,"'ilffil[:ljili:fiffi ^{rE ucou ntrie s} Herto Dwi Ariesyady, ^Tsukasa lto, Badariah Yosiyana,Marisa'Ilanclajani

L,-

The Propionate-Oxidizing Bacteria (POB) Diversity of Anaerobic Digesters

in

Tropical Environment as Revealed by 165

rDNA

Gene Cloning

Analysis

87

I M.

Wahyu W., E. Damanhuri, Tri Padmi, Kuntjoro A. Sidarto

Study of Computer E-Product Qwnership on Household Level

in

The City of

Bandung

95

Kania Dewi

Oxides of Nitrogen (NOx) Abatement by Photocalytic

Activity

Using

Titanium

Dioxide

(TiO2)

₁₀₂

Liza Ferina, Shaliza lbrahim, M.

Sffian

M. Annuar

Optimization of Aqueous Remazol

Brilliant

Blue R (RBBR) Decolori2ation by

Trametes sp. Pellets in Fluidized Bed Reactor

(FBR)

108 Maria Antonia N. Tanchuling, Harold M. Aquino, Dennis C. Ong, Augustus C. Resurreccion

Removal of Zinc and Lead Ions from Aqueous Solution Using Coco-Peat as

Sorbent

Material tl4

Mochammad Chaerul, Adi Susangka

Multicriteria

Analysis

for

Selecting

Municipal

Solid waste

Composting

Technology _lZ0

"/'

Nadia Paramita, Prayatni Soewondo

Strategy

for

Sustainable Domestic Wastewater Onsite Facility Improvement

in

SIum

Area

through PNPM

Mandiri

^(Case study: Sadang Serang Area-Bandung,

westJava)

. ^-.,-r--

^ ,?, n /"^

^d,

. rr

⁷

., . Ut

Nguyen Thi Van, Dr. Maria Lourdes Dalida, Prof.Dr. Shiro Yoshikawa, Prof, Dr.

Huynh

^\--

Trung

Hai

Adsorption of Metallic Ions onto Chitosan coated bentonite:

Equilibrium

and

I(inetic l3l

Norzila Othman, Noor Hana Hussain, Ahmad Tarmizi Abd Karim, Suhaimi Abdul-Tatib Isolation and Optimization of Napthalene Degradative

Bacteria

140 Nur Atiqah Ismail, Shaliza lbrahim, Mohamad

Sffian

Mohamad Annuar

Effect of Selected Physical and Chemical Parameters on Crystal Violet

Decolorization by Immobilized

Laccase l4S

P a tip arn Puny ap a I a lail, Sum e th Jun s u th o np oj

Selective Adsorption of Heavy Metals and Ionic Dyes on Silica Coated

Superparamagnetic

Particles _E2

Phetyasone Xaypanya, Augustus C. Resurreccion, Ma. Antonia N. Tanchuling, Ken Kawamoto, Toshiko Komatsu, Jiro Takemura, Per Moldrup

Linking

Methane Bmission Rate to Gas Transport Parameters

of

Final Soil Cover at Payatas Controlled Dump Site in the

Philippines

f Sg

Prihandrijanti, M., Malisie, A. Firdayati, IuI., Otterpohl, R.

Ecological Sanitation, an alternative

for

a Sustainable Domestic Wastewater

Management System in

Indonesia

₁₆₅

Puji Lestari, Yandhinur D Mauliadi

Application of PMF Model

for

Sources Apportionment

of

Aerosol Fine and Coarse Particles during

Dry

and Wet Seasons in Rural Area, Bandung,

Indonesia fiz

The _1't International Conference on

sustainable Infrastructure and Buirt Environment in Deveroping countries November zndgd zoog, Bandung,west jauJ- _rnodneiia

t

/ ^prtt

sri Komala, Agus Jatnika Effendi,

IG.

wenten, wisjnuprapto

FIux Continuity in Membrane bioreactor for Azo Dye Biodegradation Putri Andini Budiman, Sukandar

Composting Process of Sludge F rom Dairy Industry's Waste Water

Treatment

Plant

_,

Ratna Permatasari, Tri padmi Damanhuri

Greenhouse _Gas Emission Study From Pet Recycling process

of

Bottled

Drinking

Water

Setiadi T, Rachma R.M, Reinaldo V, Muhyinsyah A

Electricify Generation from _Tapioca wastewater using

Microbial

_{Fuel Cell} (MFC)

Siti Ainun, Yulianti pratama, Elvin Malino

study of Domestic packaging waste Generation case study:

I(elurahan Cigadutrg, Bandung, West Java

siti suhana sulaiman, shariza lbrahim, M. suffian M. Annuar

Dye Decolorization by peilets of pycnoporus sanguineus using Stirred Tank _Reactor

Theingi Soe, Ma. Antonia N- Tanchuling, Augustus C. Resurreccion, Ken Kawamoto, Toshiko Komatsu, Jiro Takemure, per Moldrup

Methane Concentration and Methane Emission Rate Measurements on the F,inal

Landfill

Cover at the Payatas Controlled _Dumpsite: Correlation _{of Methane}

Concentration to Field Water Content and Ground

Temperature

_z2Z

Thuy vu Kiem, Manh vu van, Maria Lourdes p.Dalida

Recovery Of Chromate

from

Chrome Electroplating _Waste Water by Anion

Exchange Resin

Method

_ZZg

Vu Minh Trang, Analiza p. _Rollon

Removal of reactive azo dye

in

a sequencing Batch Reactors:

Effects of Total Cycle Time _and Anaerobic:Aerobic Retention Time

Ratio

23g POSTER SESSION PAPERS

Adyati P. Yudison, Driejana, M. Irsyad

Correlation of SO2 Bmission

from

Open Burning

With

Solid Waste

and

Burning Characteristic

₂₄₄

Christy Yohana Wulandari, Sukandar

Hazardous Waste Generation and Composition of Private Dental practices

(Case Study Bandung

City)

_ZS0

Eka Wardhani, Etih Hartati, Luffia

Fitri

^yusuf

rdentification _{of Clean} Production Opportunities at Textile Industry

of

"PT X"

in Indonesia

Emenda Sembiring, _Vilas

Nitivattananon

²⁵⁸

Model Formulation

for

a Preliminary Decision-Making _Tool

for

a Regional Solid Waste Management in Developing

Countrie, "

Kania Dewi, Ivan Yosafat Siahaan 264

Optimation of Carbon Dioxide Abatement

in

Carbonation Reactor

utilizing

solid waste of

carbide

werding

process

Nadia, R., Mohd. Bakri, 1., Khairul Nadiah, L 270

178

184

196

242

208

215

vl

Transport and Trade of Electronic Waste in Malaysia: Aspects and rssues 276

'r

*#ffifo

'u*"u'"

"offi

;ff;qfif1f,ffifrji,rru;;g,co,rnbies Nazaitulshila R., Zulbahri R.H

The Socio Economic Effect on Municipal Solid Waste Generation

and Composition:

A

Case Study in Kuala Terengganu, Besut And

Kemaman

2g0 Siti Ainun, Harrie van Bommel

Study of the Criteria Assessment In The Irnplementation _of Packaging Waste Management System Related to Integrated Product Policy and Extended

Producer

Responsibilify

_2g6

LIST

OF PRESENTER

NDEX OF'AUTHOR

ACKNO\mTEDGMENT

International _{November, 2-3,} conference _on sustainable Infrastructure _and Built Environment in Developing countries 2009, Bandung, West Java, Indonesia

/,sBN 9 7 8-9 7 9_9 8 2 7 8_ 2 ^_

I

The

Application

of

Bioemulsifier _Produced

by Azotobacter

vinelandii in Bioremediation oil-contaminated _soir

water&waste#f:3,rr"t*;":,#?l*!;,";l;ff':?'aiftEllHH:"ta,Engineering,

Institut Teknologi Bandung Ganesha 1 0 Bandung-West lava,Indonesia

*Corresponding

author : j atnika@indo.net.id Abstract

Emulsification properties of the bioemulsifier produced by Azotobacter vinelandii were stable at wide range o{ tem4erqture,

pH

and

$tnity indicatini

advantages

over

its applications in the bioremediation of oil-contaminated soil

ind

othen lurposes in

oil

industry. The addition of this bioemulsifier in a broth culture was able to reduci the surface tension

from

^{72.67 to}

45'85

dyne/cm'

The addition of

bioemulsifier

in the

bioremediation process

of oil-

contaminated

soil

could enhance, the degradation process

by

making the^ cntde

oil

more soluble and readibl to petrophylic

bactJia b

atta,ck this substrate as indicated

with

the bacterial growth' The TPH rimoval efficiency of reactors after runningfor _about 3 days were 47'04%for reactor with addition of petropiytii.bacteria

orb, *hrrrii

in the addtion of 5%

bioemulsifier produced by Azotobotiu,

viielandii

the

,tioroi ,6"iency

reached 6Lg%. TpH removal in the reactor control was about 4.56%for

reactorcoioot

_{and 6I.g%.}

Keywords : bioemulsifi er, Az o to bacter vin e landii, TpH, removal effi ciency.

1.

Background

Bio-emulsifiers have been applied

widely in the

environmental protection, crude oil recovely, agriculture, mining, health care and food-processing industries (Desai and Banat, 1997)' Bioemulsifiers have special advantage over ihe

chem[al

surfactants, such as lower toxicity, higher biodegradabiiity, better

"rru"iro*ental

compatibility, higher foaming, high selectivity and specific

1997; Makkar and

Cameotra activity at

,

^1997a). extreme temperatllre, pFf and

As

environmental

compatibility

salinity (Desai and becoming Banat,an

increasingly

important factor in the

selection

of industrial

chemicals,

the use of

Bioemulsifiers in environmental application, such as biodegradation and dispersion of oil spill is increasing (Banat, _1995). m aaaiiion, Bioemulsifiers cou-ld also be used for others pu{poses in petroleum industry such as enhancing

oil

recovery process and the transportation of crude

oil'

other possible application fields

ur.

in the food, ctsmetic and pharmaceutical industries.

In these industries, most Bioemulsifiers are used as emulsifiers. The largest fossible market for Bioemulsifier is in the

oil

industry both for petroleum production _and for bioremediation

of oil

contaminated sites. The application

of tioemulsider in

the bioremediation process r'vould accelerate

the

nafural biodegradation processes

in

contaminated environments.

Bioemulsifiers are a structurally diverse group

of

surface-active moleculer

rv"rt

"rizil;

microorganisms. These molecuies reduce-r,rriu"" and interfacial tensions in

both

aqueous

solutions

and

hydrocarbon mixtures. Bioemulsifiers

activities can be

determined by measuring the charge emulsion' The surface tension

in

surface and interfacial tensions, stabilizatiorr at the airlwater and oil/water interfaces can

or

easily destabilization be measured

of with a

tensiometer. The surface tension

of

distilled water

is 72;*Nd, ;iia

uaaition

of

surfactant lowers this value

to

30 mN/m (Desai and Banat,

rggl). A1

emulsion

is

formed D: WASTE ENGINEERING AND MANAGEMENT

I

69

when one

liquid

phase

is

dispersed as microscopic droplets

in

another

liquid

continuous phase. Bioemulsifier may stabilize (emulsifiers) or destabilizer (deemulsifiers) emulsion. The emulsification activity is ^assayed by the ability

of

the surfactant to ^generate turbidity _{due to} suspended hydrocarbons such as

a

hexadecane

or

Kerosene

in

an aqueous assay system"

Hydrocarbon

(CxHy)

degradation

in soil has been

extensively studied among many researcher. Degradation is dependent on presence in soil of hydrocarbon-degrading species

of

microorganisms, hydrocarbon composition, oxygen availability, water, temperature, pH, and inorganic nutrients. The physical state

of

CxFIy can also affect biodegradation. Addition

of

synthetic surfactants or microbial emulsifier resulted

in

increased mobility and solubility

of

CxHy, which is essential

for

effective microbial degradation. Use

of

bioemulsifier

in CiUy

degradation has produced variable results

i.e

the fungus Cladosporium resiuae,

grorl

on alkane mixtures, produced extracellular

fatty

acids and phospholipids, mainly dodecanoic acid and phosphatidylcholine. Supplement

of

the growth medium with phosphatidylcholine enhanced the alkane degradation rate

by

30%. Emulsan, stimulated aromatic mineialization by pure bacterial cultures, but inhibited the degradation process when mixed cultures were used. Naphthalene was utilized in the first phase of CxHy degradation; other

oil

components

were

degraded

during the

second phase

after the

surfactants produced

by

concemed microorganisms lowered the interfacial tension. Addition of Bioemulsifiers, such as bacterial population was enhanced. Surfactants have been studied for their use in reducing viscosity

of

heavy

oils,

thereby facilitating recovery, transportation, and pipelining. _Emulsan,

a

high molecular weight lipopolysacharide produced by

A.

calcaocetiiui, _{has been} proposed

foi

a

number of applications in the petroleum industry such as to clean

oil

and sludge

fro*

barges and tanks, reduce viscosity

of

heavy

oils,

enhance

oil

recovery, and

stabiliie

water-in-oil emulsions

in

fuels. The specific solubilization

of CxHy

was strongly inhibited

by

EDTA which was overcome by excess Ca*+. It was concluded that specific solubilization of CxHy is an important mechanism in the microbial uptake of CxHy (Karanth et al., 2005).

2.

Methodology

Microbial Source & Medium

Azotobacter vinelandii and petrophylic bacteria were obtained from the culture collection of Environmental Biotechnology Laboratory-Environmental Engineering Department, Institut Teknologi Bandung, Indonesia. These microorganisms were -uitrtuitted ^{at qo} C on mannitol enrichment agar slants containing

(glL):20

mannitol, 20 yeast extract,

20

tryptone, I

5

agar.

Sub-cultures were made to fresh agar slants every 2 month to maintain

viabiiiiy. All

cultures were grown aerobically

in Minimal

Basal Medium

(MB)

which contained the following components (g/L)

of

distilled water: KzHpOq 1.5; KlI2pO4 0.5; MgSOq0.2; _OIH4)2 SO+ 0.5;

glucose 5. 10ml Trace Element solution was added per liter of

MB medi"*, ih"

"o*position

of this

trace element

(g/L) is

Na2EDT

Az.Hzo 12 ;

^Fesoa

^.7Hro z ; ^caClz 1

;

znsoq.TH2o 0.4;

NaSoa

10;Mnso4.4H2o 0.4;cuso+.5H2o _0.1

; Na2Moo4.2H2o 0.s.

Microbial _Growth

50 ml of MB was inoculated with 0.5 g of oil contaminated soil and oil sludge in a 250 ml erlenmeyer flask and incubated

in

an orbital shaker at 150 {pm. The inoculated flasks were incubated at room temperature

for

several days

until visibll

growth was observed. 10 ml subcultures were used subsequently to inoculate fresh batches

of

Ndg medium which added with

I

^o/o crude oil. The enriched flask were incubated until visible emulsification of crude oil layer was noticed. This procedure was repeated several times

to

ensure the capability

of

microbial culture in emulsifying crude oil in the medium.

B io emulsifi er Pro ducti on

To

produce crude bioemulsifier, an inocula was prepared

by

transferring the bacterial colonies into a MB media with selected carbon soruce,

ind

the culture was incubated at room temperature

in

a shaking incubator at 150

rpm

for 24 h. Then a

MB

media containin g 2%

inoculums and 2o/o carbon source was incubated at room temperature (280 C) under aerobic condition

in

a shaking incubator

at

150 rpm

for

48

h to

obtain the highesf microbial and

trL /

INEI^IECVNVI^I CNV DNTaIINIDNa EISV/Y\. :q

ul (svr)

eluuoqdlns euezuog

I{{IY

^{reeur-I sB} IIo^\ ^se

(scs)

eleqdlng _1,{cepoc urnrpos

o{{

luBlcsJms ^c1pq1u,{s

ol

peJeduloc setpedo;d eceyms qEF{ .{1en4e1eJ pe^\oqs (sregrspuroorg opruJ) _Worq ea:J IIec Jo uolsuo} ^eceJrns eq} lerll uees eq plnoc 11 'enoqe

I ^olqq

^rrrord

7,007, 'uu1qsn1 pu? golsrrrlc

uorlBqncur Jnoq gt Joue lueuroJnseehl*

eleuogdlns euezueqfl{l? reorrrT LV

e007 'uqqsnl

pu€

golsrnlJ ^ot

Z00Z 'ewqsnl

pu'

IloprnlJ

^o€

0z uoe^u

eprruoJq ruruuouruelr(qleruuq,{1a3 (regrslnrueorq epruO) _Vtorq ee+ IIoJ

eluqdlns 1.(cepop urnJpos %I wnlpelN glr{orD ellJel s lsepenby ellrels

*BleP qcJuossu

€lep qcrsesou

€l?p gcrBoseu Blsp rlcrBeseu

98'9t br'gg

L9'7,L T6'TL )oSZ lB (tur/au^p

uolsuol OJBJJnS

secueJoJal -reqlo qll/K peruduoc upuoPu!^ 'V WIm uollelncoul ^rQUu pue eroJeq rllorq go ,grnltce ecgJrns

I ^alqul

:siKolloJ se

'l

olqsJ, w uoes eq plnoc socueJeJeJ JeIIlo o1 peredruoc trpuDlaury rapDqopzv ruo{J pecnpord regrsFrueolq

eql

uoluppe eql re$B suorlrpuoc emllnc umrpo111 I€JoAes Jo setgedo:d ecu;-lng 'setgedord enrlce-olsuel pooE n

ieq

orln inq .:egrsFruo _pooE

n d,rr"q rtuo

1ou sezn'{Jo/K slr{l

uI

perynls suolllpuoc eq} Jepun

upunpurt'tr

ruor; polulosr rogrslnrueolq

otil

^'dllulles pue _11d 'e;nleredulel;o e8uu: epltr 18 olqe5 ere.& Jegrslnueorq eqlgo ssrgedord

uollecglslnlufl

'sseulotq

Ep _8'89 ol dn

^p1er.'( sB/K Jeglspruaorq

Jo

uotlc.,pord

oql

_l€ql

punoJ sB/K

lI

suoltrIpuoc eseql Jepuo 'llpuDlau!^ Jap^qotozv tJtoJJ JogrslnuaolqJo uollcnpo:d

eql pellql{q

uol}ur}uecuoc gEU

uI

suol _l?}oru

Jo

^ecuoserd

eq1

'uottcr,poid regnlnrueorq

eql

_lceJJB

ol

punoJ se/tr suolluc

Plolu Jo

^uonrppu

erdl

'pesseJcul ss/r\ uorlBJluecuoc ueEorlru

uoq^\

poseorcep Joglslnrueolq

Jo

uolicnpord

",tt

.rtpuoput^

rap7qorczy

.(q pecnpord JaglsFtuoolq Jo uollcnpeJ egt ^.,(q pol\olloJ sn^ q1^or8

fp"

^go

"rn"r"o,

^eq]

.,{.rur1uoc

uI 'tll

\or8

lec

3o

pler,

reqEg ur pellnser uolle4uecuoc ueEorlp

,.!atg

^.olpJlns snorred Jo ocuosqB eql

q

^pue alsJlns uInIsouBeIN

lF

^{Z'O 'e1er1ru} runruoruruy I1ESZ'Oqlr/!\ polu"*"1dOi-tt seK rlpuoputt raPoqoQzv Jo t{tmor8 ^er{l ueqt\ pelolqce sBA\ JelJrslnruoorq Jo uor}BJ}uocuoc

qEIII{

'pelpqs

_sB^\

!!puqau!^ rapoqorczv .(q

uorlcnpord regrspruoorq

pus qlr\or3

uo

Eur1ce3Je srolseJ snolreA 'uolleluouusJ _Jo rnoq

zL

^JatJe p"rr,pord ilieruqoo'ss r regrsFruoorq oql lBt{} pe}eclpul ,(pn1s snorrrerd eql urog I1nSaU 'roded Joqlo ur polu"serd puu

fp"rr

rr,ot,t"rd egl ut pezuepBrel{c _ueoq pBq ttpuqtau^ 'tap2qopzv

tuo{

pecnpord sregrslnue-olq

aql

uorssncsl(I

T llnsou

^.€

'Cog, ls peleJodeng sB/K arrexeH oql roUu EuqErem sern ged lueleuadns er1l'salnurur

0I

^roJ

*Ar

_Oosg 1e Eur3rUUlrs

iq

polcuJlxe olgnop ust{l se^\ omlxFu

slql

^'euexeH _{Fu S} q}l/l\ pexllu sern eldues Jo Iru S

V

uoueDqxg uoqrac orpt11 wnap4ad IDtoJ 'luo^Ios eql e^oluer ol poleJodene pue lcnpord ragtslnur?olg opruc peJoloc-u^\oJq snocsr1 _B Eurpler.(

Slseu uouoq-punoJ

3 ol

perJeJsrreJl

set\

aseqd cpe8ro

orIJ

.erqsroduJel

ruooJ lE lue1los uuoJor.olqc € qU^\ dels uorlce4lo JegIsFIuooIq oq]

rq

pernollog 'uollnlos clrolqcorp.(q

ld f

e

u

^{pe^losslp} ue{} sB/t\ lue}eulodls agl 'uotleEnglrluec raUV ^'QOOZ ''le le ee/r\sluoqtunsruo6) sregrsFrueolg

eql Jo uorleildrcsrd eplduroc eql roJ Joi w

lqErrue,ro

uel

^sg,y\

lueluuedns peUlplce eql puu 'uoltnlos ^pIcB cFoplcorprq

w fHotr.,

0.2

Hd ol

uortecglplce

{q

^pe1ue4 requru sB/K truelerue{ns peu*}qo

eqJ

^'sllec _{Ielrelceq eql} e^oruor or ropro ur urru 0E roJ uldr 0oo'€I ^puB _{Jov w} peSryrrluec set\ uor1.,1os eql 'leqtr reUV 'uorlg4uocuoc tu'lceJirns

lowering the

surface tension.

This result

suggest

that

bioemulsifier

from

Azotobacter vinelandii have a potential function as emulsifier/surfactant. Therefore, this bio-emulsifier could

be utilized

as surfactant

in

order

to

improve the biodegradation

of

petroleum

oil

process. The main physiological role

of

bioemulsifiers is to permit microorganisms to grow

on

water-immiscible substrates

by

reducing

the

surface tension

at the

phase boundary.

Therefore, this role makes the substrate more readily available for uptake and metabolisms. In addition

to

emulsification

of

the carbon source, they are also involved

in

the adhesion

of

microbial cells

to

the hydrocarbon. The cellular adsorption

of

the hydrocarbon-degrading microbes

to

water-immiscible substrates, and

the

excretion

of

surface-active compounds together allow growth on such carbon sources. In general, negatively charged bioemulsifiers

would inhibit the

process, whereas positively charged bioemulsifiers promote microbial adhesion

to

hydrophobic

phases.

Furthermore, bioemulsifiers have been shown

to

be involved in antagonistic effect towards other microbes in the environment.

Bioemulsifiers are also used

in

emerging technologies

like

microbial remediation

of

hydrocarbon and crude oil-contaminated soils. Hydrocarbon contaminants are removed from the environment, primarily, as a result of their biodegradation, which is performed by-native

microbial

populations. Such biodegradation

is known to be

time-consuming

and

new technologies have been developed;

for

example bioaugmentation, biosurfactants and etc.

While, the effectiveness of enhancing hydrocarbon degradation through addition of microbial inocula of non-indigenous populations (bioaugmentation in essence) has been ambiguous, the use of a combination of new approaches has shown much success. For instance, the addition

of

bioemulsifiers

help to

stimulate

the

indigenous

microbial

population

to

^degrade

hydrocarbons at rates higher than those which could be achieved through addition of nutrients alone.

A

good example

of

this is the use

of

rhamnolipid from Pseuclomonas aerugino.sa to remove large quantities of

oil

from contaminated Alaskan gravel a decade back, in the Exxon Valdez oil spill.

A

laboratory scale

of

bioemulsifiers enhanced biodegradation

of

crude

oil

was conducted.

The effects

of

addition of bioemulsifiers from Azotobacter vinelandii

in

the biodegradation process are shown in Figure 1a-le.

1

.8 .b .4 .2 0

2A 30

^4A

Time {hour}

**"*Growth

6

4.^

5 ab?

,T, i*

0

2A 30 4A

⁵⁰

Time (hour)

**e"*Growth

s

o0

s

-o

80

a)

b

4*

10

r(L

I

'l- o

b)

1.8

di b.s S.o

f;o fu.g

10

Continued

60

c)

1.5

$1 ^.2

Ho.s

8o.e

go bo.s

{ 2A_ 30 4A

⁵⁰

Time (hour)

**S* _Growtlr

60

b

4^ *.

2r,

_(L

2A 30 40 ⁵⁰ OF Time {hour}

*-s,*Growth

60

I o0.8 Po.e

$o.c

f;o 84.2

0 {0 20 30 4A

⁵⁰

H:."[]?,i"

Figure

I

^The biodegradation of petroleum oil in the presence of bio-emulsifier produced from Azotobacter vinelandii (a). control,

6). ox,

^{(c). I .5oA, (d).3%,}

(").

sy"

Figure

I

showed the perfofinance

of

Petrophylic bacteria

in

degrading crude

oil.

There was no considerable decrease

of

Total Petroleum Hydrocarbon (TPH) concentration

in

the control reactor where was no addition of petrophylic bacteria

in

it. The reduction of TpH in this reactor was most likely due to physical influences such as temperature and volatilization of low molecular weight of hydrocarbon. In contrary, there was a significant decrease of TpH concentration that occurred

in

the reactor where petrophylic bacteria were added. These results suggested that the bio-augmented bacteria could degrade TPH significantly compared to control reactor. Physical and chemical transformation might also occur

in

the process as

shown

in

reactor control. However, based on research result the biological transformation seemed

to

dominate the process. The growth

of

bacteria

in

the reactor was also observed during the process. This related to the biological transformation during the process in which the number of bacteria could be maintained during the degradation process. It was proved that

the

addition

of

bio-emulsifiers increased the biodegradation efhciency

of

petroleum

oil

removal markedly as showing in Table 2.

0 10

1

o0.B Po.e

d) 50.+ _bo.z

tso {

0 10

e)

6 5 4 "^,SO

1l o-*

9T, i*

0

D: WASTE ENGINEERING AND MANAGEMENT

I

⁷³

Parameter

Table 2 TPH removal effici in Presence of Bio-emulsifier

Initial

TPI{

^{Final TPH} Efficiency Control Reactor (- Petrophylic (P), - Bioemulsifier)

Reactor

I

⁽⁺ P, - Bioemulsifier) Reactor 2 (+P,

*

Bioemulsifier l,5o ) Reactor 3 (+ P,

*

Bioemulsifier 3%)

5

5 5 5

4.757 2.648 2.296 2.042 1.910

4.86 47.04 54.08 59.16 61.80 Reactor 4 (+ P,

*

Bioemulsifier 5%)

(-): without; (+): with

It

was evidence

that

bio-emulsifier

could

enhance

the

biodegradation process. The

limiting

factor

in

the degradation

of

hydrocarbon and other

PAI{

is their insolubility, thus decreasing the efficiency and rate

of

degradation. This limitation can be overcome either by addition'

of

surface-active compounds (surfactant)

to the

growing culture, thus making hydrocarbons more water-soluble and available for the cell to degrade, or by production of its

own

surfactant

by the

organism

to facilitate the

uptake process. Bio-emulsifiers are

'amphiphilic' molecules consisting of both hydrophobic and hydrophilic domains giving them the characteristic property of organizing at interfaces

of

different degrees of polarity zuch as

oil/water

or

airlwater and helping

to

lower the interfacial energy/tension.

i6.

addition

of

bioemulsifier could enhance the degradation process by making the crude

oil

more soluble and readily to petrophylic

in

attacking the substrates as indicaied

by its

growth. The TpH removal efficiency

of

reactors after running

for

about 3 days were 47 .04%

for

reactor with addition

of

petrophylic bacteria

only

and 61.8%

for

reactor

with

addition

of

petrophylic bacteria and 5o/o bioemulsifier, whereas the removal efficiency

in

the reactor control was 4.86% as shown in Table 2.

4.

Conclusion

The bioemulsifier produced by Azotobacter vinelandii exhibited significant emulsification properties. The effect

of

addition

of

bioemulsifier

in

the crude

oil

biodegradation process indicated the enhancement

of

both degradation process and growth

of

petrophylic bacteria.

TPH removal efficiency increased from 63.43% up

to

82.04% as the addition of

bior*factant

to the process.

5.

Acknowledgement

The authors gratefully acknowledge the financial support received from Indonesia Toray Science Foundation ITSF, 2007 and ITB for providing the facilities for this work.

6.

References

Banat,

I.M.

1995.

The

isolation

of a

thermophilic biosurfactant producing

Bacillus

sp.

Biotechnol. Letter. 1 5: 59 I-594

Banat,

I.M.,

R.S. Makkar and S.S. Cameotra. 2000. Potential Commercial

Microbial Surfactant. Appl Microbio Biotechnol, vol. 53:495-50g Applications

of

Desai, J.D. and I.M. Banat. 1997. Microbial Production of Surfactants and Their Comrnercial Potential. Microbiol. And Molecular Biol. Review. p.47-64.

Karanth, N.G.K., P.G.Deo and N.K.

Veenanadig.

2005. Microbial Production of

Biosurfactant and Their Importance. http://www.ias.ac.in/currsci/

jull0/

articleslg.htm Makkar, R.S and S.S. Cameotra. I997a. Biosurfactant production

by

thermophilic Bacillus

subtilis strain J.Ind. Microbiol. Biotechnol. 18:37-42

Makkar, R.S and

S.S. Cameotra. 2002.

Effect of

various

nutritional

supplements on biosurfactant production by a strain of Bacillus subtilis at 450C. Journal of stufactants and detergents 5:1 1-16

t920

rsBN 1?8-1?1-16e?8-A-1

llil

illllilt

lililt lll ill

llllll

ell789799ll8?7821il