BIOCONVERSION TECHNOLOGIES

FOR INDUSTRIAL BIOPROCESSES

Bioprocess Considerations in Using Microbial
For Conversion into value added products

a) Advantages and disadvantages of

bioconversion
b) Type of bioconversion approaches: e.g
solid-state and liquid-state
c) Important factors for bioconversion
d) Application of microbial in bioconversion
technologies utilizing waste as substrate for
production of value-added product
-BIOETHANOL
 Types of Bioconversion
SSF - Fermentation of solid substrates at low moisture levels or water
activities. The water content of a solid mash in SSF varies between 40%-80%.

Liquid state fermentation (Submerged) - The water content of a typical

submerged fermentation is more than 95%.

SSF used for the fermentation of agricultural products or food ( rice, wheat,
barley, corn and soybeans).

The unique characteristic of SSF: Operation at low moisture levels (provides

a selective environment for the growth of mycelial organisms (molds) –
producing extracellular enzymes on moist agricultural substrates.

Examples: SSF are used widely in Asia for food products (tempeh, soy sauce
fermentation)
Advantages over submerged:

1) Small volume of fermentation mash or reactor volume –

resulting in lower capital and operation costs

2) A lower chance of contamination due to low moisture

levels

3) Easy product separation

4) Energy efficiency

5) The allowing of the development of fully differentiated

structures, which is critical in some cases to product
formation
 Major disadvantages:
1.The heterogenous nature of media due to poor mixing
characteristics – results in control problems (pH, DO
Temperature) within the fermentation mash (to eliminate these
control problems-fermentation media is usually mixed
(continuous or intermittent)
2.The concentration gradients may not be eliminated at low
agitation speed especially for large fermentation mash
volumes
3.Mycelia might damaged at high speed agitation speeds
–rotation drum fermentor can be used and rotational speeds
needs to be optimized.

Examples of SSF:
1.Bacterial ore leaching
2.Fermentation of rice in a packed column
3.Koji process employ molds (Aspergillus, Rhizopus) growing on
grains or foods (soybean, rice, wheat)
 SOY SAUCE PRODUCTION

Soy Beans

Pretreatment of soy beans (cleaning,

soaking, cooking and draining)
Fungus culture
(Aspergillus or
Rhizopus) + Mix
wheat flour

Incubated in koji room (48 hours)

Koji
Brine solution

Incubate Moromi (2 months)

-external added of bacteria/yeasts for
flavor producers (8 to 12months)
 Pressed to
Extraction Residue recover liquid
soy souce.

Filtrate
Additives

Pasteurization

Packaging

Soy sauce @ Kicap

Indonesian soy sauce

Chinese soy sauce Japanese soy sauce @ Shoyu

 Koji Process – the production of
enzymes by fungal species
Koji process- the primary stage is an aerobic, fungal solid-state
fermentation of grains.

Moromi - The second stage is anaerobic submerged fermentation with

a mixed bacterial culture

Fungal amylases are produced by SSF of wheat bran by A. oryzae in a

rotating-drum fermentor.

Pretreatment of wheat bran with formaldehyde and initial pH 3.5-4.

Perforated pans, rotating drums or packed bed with air ventilation are
used for SSF.

Other enzymes (cellulase, pectinase, protease, lipase) can also be

produced by koji fermentation.
 Some secondary metabolites such as antibacterial agents are produced by
Rhizoplus and Actinomucor species by koji process.

Other Examples:
-Aflatoxins are produced by SSF of rice (40% moisture) by A.parasiticus.
-Ochratoxins produced by Aspergillus species on wheat in a rotary-drum koji
fermentor.
-Microbial degradation of lignocellulosics can be accomplished by SSF for waste
treatment purposes
-Spore formation is obtained by SSF

Major process variables in SSF:

1.Moisture content (> 60%-solid substrate become sticky and form large aggregates,
affect the metabolic activities of cells, different cell-substrate system will created
different moisture level, optimum moisture level is 40±5%
2.Inoculum density
3.Temperature
4.pH
5.Particle size – small enough to avoid oxygen-CO2 exchange/other nutrient
transport limitation. Porosity can be improved by pretreatment
6.Aeration/agitation
 How to perform SSF:
i.Pretreatment :
most natural substrate need pretreatment-substrates is
susceptible to mycelial penetration and utilization

ii.Treated with microbial agents and steamed in an autoclave

iii.Preparation of nutrient media, pH adjustment

iv.Adjustment of moisture level

v.Inoculation of fermentation mash

vi.Realized in a controlled-humidity air environment with air ventilation

and agitation
 Application of microbial in
bioconversion technologies
utilizing waste as substrate for
production of value-added
product

–FUEL BIOETHANOL-
 Evaluation of feedstocks
Bioethanol can be produced from raw materials:

1) Sucrose-containing feedstocks
Sugar cane, either in the form of cane juice or cane molasses, is the most
important feedstock utilized in tropical and sub-tropical countries for producing
ethanol.

2) Starchy materials
To produce ethanol from starch it is necessary to break down the chains of this
carbohydrate for obtaining glucose syrup, which can be converted into ethanol by
yeasts. This type of feedstock is the most utilized for ethanol production
in North America and Europe. Corn and wheat are mainly employed with these
purposes. In tropical countries, other starchy crops as tubers (e.g. cassava) can
be used for commercial production of fuel ethanol.

3) Lignocellulosic biomass
Lignocellulosic biomass (a complex comprised of several polysaccharides) is the
most promising feedstock considering its great availability and low cost,
but the large-scale commercial production of fuel ethanol from lignocellulosic
materials has still not been implemented.
Important factors in choosing the suitable biomass for use as a raw
material for fuel bioethanol production:

•As long as the particular biomass contain of cellulose and hemicellulose

component, it can be converted into fermentable sugars by hydrolysis and
can be fermented to produce bioethanol. Low lignin content in the biomass is
favourable as it is recalcitrant to degrade.
•The potential biomass must be abundant, reproducible resources and
available from season to season. For example, Malaysia generates a large
amount of biomass from palm oil industry annually. The oil palm empty fruit
bunches which contains of cellulose and hemicellulose is produce in huge
quantities are still underutilized. Therefore, this material is suitable to be a
feedstock for the conversion to variety of products attractive.
•The particular biomass should be inexpensive to make the overall process
economic feasible and should not be highly volatile, which can highly affect
the production costs of bioethanol. If the biomass is a domestic resource
which is not subject to world price fluctuations, it will be better to be used as
raw material.
•The materials also should be non-food biomass so the issue on food price
has not been raised and the people hungry are not going to happen.
 Conversion of lignocellulosic
biomass to ethanol
Overall fuel ethanol production from lignocellulosic biomass
includes five main steps:
(1)biomass pretreatment
(2)cellulose hydrolysis
(3)fermentation of hexoses
(4)separation
(5)effluent treatment

Furthermore, detoxification and fermentation of pentoses

released during the pretreatment step can be carried out.
BIOMASS PRETREATMENT
Structure of Lignocellulosic Biomass
 Why do we need biomass pretreatment
Lignocellulosic biomass needs pretreatment due to
the existence of various physical and chemical
barriers that greatly inhibit their susceptibility to
bioprocesses such as hydrolysis and fermentation.

These barriers were due to the close association of

the three main components of cellulose,
hemicellulose and lignin, which differ significantly
from plant to plant.

In short, pretreatment is necessary for effectively

breaking the biomass recalcitrant characteristics
 PURPOSE
•Alter or remove structural and compositional barriers
•Open up the cellulose or hemicellulose structure to
hydrolysis in order to improve the rate of enzyme
hydrolysis or increase yields of fermentable sugars
•Increase the susceptibility of cellulose to enzymatic
attack.

In general, an effective pretreatment is characterized by

several criteria which include avoiding size reduction,
preserving hemicellulose fractions, limiting formation of
inhibitors due to degradation products, minimizing
energy input and being cost-effective.
PHYSICAL BIOLOGICAL
PROCESS PRETREATMENT
Size reduction • Use microorganisms
- Increase the accessible • Fungi use to treat biomass
surface area and enhancing enzymatic
- Reduce cellulose hydrolysis
crystallinity • 5 week operation time
- Method : grinding, • Low energy requirement
chipping, milling
PHYSICO CHEMICAL PROCESS
CHEMICAL PRETREATMENT
 Hydrolysis Step
• Hydrolysis is the second step, after pretreatment process, whose
objective is to produce monomer sugar such as glucose and xylose
from cellulose and hemicellulose.
cellulose glucose
(C6H10O5)n + H2O → nC6H12O6
hemicellulose xylose
(C5H8O4)n + H2O → nC5H10O5

• The sugar molecules will be degraded to ethanol in the fermentation

step.
• There are two types of hydrolysis:
1. Acid hydrolysis
2. Enzyme hydrolysis
 1. Acid Hydrolysis
•Mainly produces xylose from xylans (hemicellulose).
•Xylan is more susceptible to hydrolysis by mild acid
treatment due to its amorphous structure compared to
cellulose, which needs severe treatment condition for
its crystalline structure.
•Acids can penetrate lignin without any preliminary
pretreatment of biomass, thus breaking down the
cellulose and hemicellulose polymers to form individual
sugar molecules
•There are two types of acid hydrolysis:
1.Dilute acid hydrolysis
2.Concentrated acid hydrolysis
 1.1. Dilute Acid Hydrolysis
•Hemicellulose fraction is depolymerized at lower
temperature than cellulosic fraction
•Dilute sulphuric acid is mixed with biomass to hydrolyze
hemicellulose to xylose and other sugars
•Involves two stages:
1.Performed at low temperature to maximize the yield
from hemicellulose and is conducted under mild process
conditions to recover the 5 carbon sugars
2.Higher temperature stage is optimized for hydrolysis of
the cellulose portion and is conducted under harsher
conditions to recover the 6 carbon sugars
 1.2. Concentrated Acid Hydrolysis
• Provides a complete and rapid conversion of
cellulose to glucose and hemicelluloses to
5-carbon sugars with little degradation
• Reaction times are typically much longer than
dilute acid hydrolysis
• Uses up to 70% sulphuric acid at 313-323 K for
2-4 hours
• Low tempretares and pressure will lead to
minimization of sugar degredation
 Comparison Between Dilute- and
Concentrated-Acid Methods
Advantages Disadvantages
Concentrated-acid • Operated at low • High acid consumption
temperature • Equipment corrosion
• High sugar yield • High energy consumption
for acid recovery
• Long reaction time (2-6 h)
Dilute-acid • Low acid consumption • Operated at high
• Short residence time temperature
• Low sugar yield
• Equipment corrosion
• Formation of undesired
by-products
 2. Enzyme Hydrolysis
•Enzyme hydrolysis is carried out by cellulase or
hemicellulase enzymes which are highly specific to
produce reducing sugars such as glucose
•Carried out in mild condition (pH 4-8 and 45-50 C)
•Advantages:
✓Low utility cost
✓Does not have corrosion problem
✓Lower energy consumption
✓Produces high yield of pure glucose with low
formation of by-products
 2.1. Cellulase Enzymes
• Endoglucanase: Proposed to initial attack randomly
at multiple internal site in the amorphous regions of
cellulose fiber opening-up site and create free
chain-ends for subsequent attack by exoglucanases
• Exoglucanase or cellobiohydrolases degrade the
chain ends further and release the cellobiose
molecule. Major component of cellulase system and
hydrolyze highly crystalline cellulose
• β-glucosidase: Splits cellobiose to two glucose
molecules
Synergistic cellulase actions on cellulose
(Source: (Dimarogona et al., 2012)
 2.2. Hemicellulase Enzymes
• Degrade more easily than cellulases due to lower degree of
depolymerization and their amorphous nature
• Hemicellulolytic enzymes are more complex and hemicellulose
contains different sugar units
• Examples of enzymes
✓ Xylanase: hydrolyze β-1,4 bond in xylan to produce xylooligomers
then hydrolyze by β-xylosidase to yield xylose
✓ Β-mannanase: hydrolyze mannan-based hemicellulose and liberate
short β-1,4-mannooligomer then hydrolyze by β-mannosidases to
produce mannose
✓ Feruloyl esterase: Helps the release of hemicellulase from lignin and
renders the free polysachharide product more amenable to
degredation by the other hemicellulases
 The benefits and limitations of using non-chemical
methods for pretreatment of biomass

The benefits of using non-chemical methods for pretreatment of biomass

are:
•Capital and operating costs are low compared to chemicals
pretreatment due to no chemicals have to buy.
•Operational is easy to handle as no explosive and highly inflammable
substances is used.
•No recovery cost of chemicals, sugar yield and removal of chemicals
after pretreatment and hydrolysis prior to fermentation.
•Maintenance cost will be reduced due to no solid or hydrolysates that
are required to be neutralized prior to biological steps.
•No corrosion problem to the vessel compared to the chemicals
pretreatment.
•No accumulation of unwanted product or toxic substances that inhibit
the enzymatic hydrolysis and fermentation process such as mechanical
pretreatment.
•Environmental friendly and energy saving process for example biological
pretreatment as it is performed at low temperature and no chemicals
usage.
 The limitations of using non-chemical methods for pretreatment of
biomass are:

•Low Cellulose Total Conversion

Cell walls of polysaccharides are rather difficult to breakdown. The
polymers of carbohydrate and lignin exhibit differential reactivity to
thermal, chemical and biological processing. Therefore, without the
combination of pretreatment normally the total conversion of
carbohydrate would be low due to low rate of conversion to
monomer sugars and produced low yield of ethanol at the end of
the process.

•High Energy Requirement

By cutting the biomass into smaller pieces like milling could only
increase the total hydrolysis yield of the lignocelluloses by 5-25%
depends on the kind of biomass, type and duration of milling. It has
a high energy requirement and was found not economical feasible
as pretreatment.
•Inhibitors
By thermal pretreatment like steam pretreatment on biomass has a risk
on producing compounds like furfural, hydroxymethylfurfural (HMF)
and soluble phenolic compounds. These are the inhibitors in the
fermentation and the biomass has to be separated from the
condensate during the pretreatment. Therefore, additional step has to
be applied to the process and will later increase the cost of overall
process.

•High Cost
Another method like irradiation of cellulose by gamma rays, which
cleaves the β-1,4-glycosidic bonds gives larger surface area and lower
crystallinity. However this method is far too expensive to be used in a
full-scale process and also doubtful to be used in combination with
environmental friendly technologies.

•Low Rate of Pretreatment

The rate of biological pretreatment process is far too low for industrial
used and some material is lost as these microorganisms also consumed
hemicelluloses and cellulose.
 Process synthesis for
conversion of lignocellulosic
biomass to ethanol
Diagram of lignocellulosic biomass fermentation for ethanol production.

Main stream components: C, cellulose; H, hemicelluloses; L, lignin; Cel, cellulases; G,

glucose; P, pentose; I, inhibitors, EtOH, ethanol
Fermentation configuration of lignocellulosic biomass
Separate hydrolysis and fermentation (SHF)
•is the classic configuration employed for fermenting
lignocellulosic biomass to the ethanol fermentation.
•In this process, the pretreated lignocelluloses are hydrolyse to
glucose and subsequently fermented to ethanol in separate
units.
•This gives advantage to the hydrolysis and fermentation
process to be carried out at their optimum conditions.
•Pentose released from the pretreatment is accomplished in an
independent unit.
• However, there is main drawback of this process whereas the
cellulase activity is inhibited by the released sugars mainly
cellobiose.
Co-fermentation (CF) of lignocellulosic hydrolyzates is refers to the
fermentation of both five-carbon and six-carbon sugars to ethanol.

•This process oriented to complete assimilation by the microorganism by all

sugars released during the pretreatment and hydrolysis of lignocellulosic
biomass.
•This can be done by mixed cultures or single culture by yeast that could
assimilate both hexose and pentose.
•The microorganisms most commonly modified for this purpose are S.
cerevisiae and Z. mobilis to which genes allowing the assimilation of
pentoses have been introduced.
•The other approach for genetic modification is the introduction of genes
encoding the metabolic pathway for the production of ethanol to
microorganisms that are capable of fermenting both hexoses and pentoses
in their native form.
•The employ of these recombinant microorganisms allows implementing
co-fermentation processes intended to more complete utilization of the
sugars of lignocellulosic biomass.
One of the most successful methods for ethanol fermentation from
lignocellulosic biomass is combination of the enzymatic hydrolysis
and fermentation in one step process, termed simultaneous
saccharification and fermentation (SSF).

•In this process, the glucose produced in hydrolyzing enzyme is

consumed immediately by fermenting microorganism in the culture.
This is great advantage for SSF compared to SHF, due to the
inhibition effects of cellobiose and glucose to the enzymes are
minimized by keeping a low concentration of these sugars in the
media.
•The number of vessels required for SSF is reduced, resulting in lower
capital cost of the process.

•The first experience with SSF from biomass converts 70% of

cellulose into ethanol and reach of 90-95% conversion with final
concentration of 40-50g/L.
 Another mode of operation is simultaneous saccharification
and co-fermentation (SSCF) in which inclusion the pentose
fermentation in SSF.
•The hydrolyzated hemicelluloses during pretreatment and the
solid cellulose are not separated after pretreatment, allowing
the hemicelluloses sugars to be converted to ethanol together
with SSF of the cellulose.
•The SSCF process is considered to be an improvement to SSF. In
SSF bioreactor, only hexoses are converted to ethanol and
pentoses can be fermented in another bioreactor with different
microorganism.
•In SSCF process, it is suggested to ferment both hexoses and
pentoses in a single bioreactor with a single microorganism.
•Therefore, only a single fermentation step is required to
process hydrolyzed and solid fractions of the pretreated
lignocelluloses.
 In consolidated bioprocessing (CBP), ethanol with all of
the required enzymes is produced in a single bioreactor by
single microorganism’s community.
•The process is also known as direct microbial conversion
(DMC).
•It is based on utilization of mono- or co-cultures of
microorganisms which ferment cellulose to ethanol.
•Application of CBP entails no operating costs or capital
investment for purchasing enzyme or its production. Several
cellulolytic anaerobes have been isolated and characterized
for potential technology development for fuel or chemical
production by CBP of lignocellulosic materials.
•This type of activity is shown by various anaerobic
thermophilic bacteria such as Clostridium thermocellum and
some filamentous fungi including Neurospora crassa,
Monilia sp. and Paecilomyces sp.
 BIOTEKNOLOGI
Bios → hidup: Teuchos → alat; Logos →
ilmu

Penerapan prinsip-prinsip biologi, biokimia, dan

rekayasa dlm mengolah suatu bahan dengan
memanfaatkan organisme hidup dan
komponen-komponennya utk menghasilkan
barang dan jasa yg bermanfaat bagi manusia.
 Mikrobiolo
Biologi gi Biokimia
Molekular
Genetika Biologi Sel Rekayasa
Molekular Proses

Bioteknologi

Industri
Diagnosa Kesehatan Industri Kimia
Fermentasi
Industri Lingkungan dan
Farmasi Energi
Industri Pangan dan Pakan
• A Science of Many Disciplines
 PETA KONSEP
Kelangsungan Hidup Manusia

Ditunjang Oleh

Teknologi

melalui

Bioteknologi

Bioteknologi Konvensional Bioteknologi Modern

Pengolahan Bahan Makanan Misalnya

Tempe Kecap Keju mikroprotein Kultur Jaringan

Rekayasa Genetik
Contoh Bioteknologi Konvensional dan Biotek. Modern

Budidaya tanaman:
- Penggunaan galur tanaman alami yg belum mengalami
modifikasi genetik
- Budidaya tanaman transgenik yg membawa gen
ketahanan terhadap herbisida

Pengendalian hama dan penyakit:

- Penggunaan bakteri Bacillus thuringiensis alami utk
pengendalian hama
- Penggunaan galur tanaman transgenik yg membawa
gen cry dari B. thuringiensis
 Review Bidang Teknik Lingkungan

• Teknik Lingkungan (Environmental Engineering) adalah Cabang

ilmu rekayasa (engineering) dg fokus utama:
– Perlindungan Lingkungan dari kemungkinan terjadinya
kerusakan sbg akibat dari dampak negatif aktivitas manusia
• Konsep Dasar Pencemaran Lingkungan:

Sumber Lingkunga Recept

n or
-Biogenik/alamiah -Atmosfir -Manusia
-Antropogenik/buatan -Hidrosfir -Tumbuha
Manusia -Litosfir n
-Hewan
-Material
Limbah pencemar di lingkungan
POLLUTION
Classification
Air Pollution
Air Pollution
Air Pollution
Land Pollution
Causes
Effects
Water Pollution
Water Pollution
Water Pollution
Contamination

impact on
organisms
Contamination
 Sound Pollution
• caused by the noise of motor vehicles, aircraft,
the roar of the engine factory, radio / tape
recorder, which reads loud that it interferes
with hearing, roadway noise, and as well as
high-intensity sonar
• will disturb human life and can even damage
hearing permanently
 Light Pollution
• comes from the exterior and interior lighting of
buildings, billboards, commercial property,
offices, factories, roads and stadium lights
• due to light pollution, bird can not migrate to the
right place because the star and the moon that as
navigator is invisible
• sea turtles also have not come to the beach and
lay eggs as usual because of fear of the light
• astronomers can not observe and find objects in
space because of too much light that covered the
night sky
 Thermal Pollution
• waste hot water from power plant installation is
usually discharged directly into rivers, increasing water
temperature and cause thermal pollution
• aquatic animal populations will decline at higher
temperatures, only a few kinds can live at
temperatures above 40oCof animals that
• oil and petrochemical kind of polluting waters will form
a thin layer on the surface of the water that prevents
the exchange of oxygen in the water with the
atmosphere
• fish can not breathe and marine plants can not
photosynthesize cause of death in biota
 Visual Pollution
• which can refer to the presence of overhead power
lines, motorway billboards, scarred landforms (as
from strip mining), open storage of trash or
municipal solid waste
• even the trees as elements of urban green evicted
by the presence of billboards. beside as the lungs
of the city, green trees also contribute in terms of
psychology to residents of the city. too many
concrete buildings and advertisements will make
people easy to stress, because it is far from nature
Visualization of The
Pollution
Receptor

• Receptor adalah penerima akibat terjadinya perubahan kualitas

lingkungan:
– Manusia: dampak pencemaran lingkungan terhadap kesehatan
• Tragedi Minamata di Jepang akibat pencemaran logam berat
• Adanya korelasi positif yg kuat antara tingginya tingkat
pencemaran udara dg angka kejadian penyakit ISPA.
• dll
– Tumbuhan
• terjadinya penurunan produktivitas tanaman produksi
• kegagalan panen,
• adanya residu logam berat dan materi pencemar lainnya dlm
tanaman maupun buah-buahan
• dll
– Hewan
• Meningkatnya angka kematian hewan ternak
• Menururnnya resistensi hewan ternak terhadap
penyakit/hama
• Dll
– Material
• Kerusakan bangunan
• Estetika Bangunan
• dll
Pencemaran Lingkungan
Sumber Pencemar Lingkungan:
– Biogenik: sumber pencemar yg teremisi ke lingkungan secara
alamiah:
• Letusan Gunung Berapi
• Pembusukan materi padat
• Penggerusan/erosi dari hulu sungai
• Dll
– Antropogenik: sumber pencemar yg berasal dari aktivitas/kegiatan
buatan manusia yg dpt berupa materi yg mampu diasimilasi oleh
alam dan materi xenobiotik (asing bagi lingkungan):
• Proses Industri
• Pembakaran Bahan Bakar
• Aktivitas Domestik
• Eksploitasi dan Eksplorasi Sumber Daya Alam: ekstraksi
mineral, minyak dan gas
• Dll
Bentuk materi pencemar yg dihasilkan oleh aktivitas manusia dapat
berupa Limbah cair, padat dan gas.
Sumber: Zero Waste Alliance
Sumber: Zero Waste Alliance
Peranan Ahli Teknik Lingkungan ?
End of Pipe Treatment

Peranan Ahli Teknik Lingkungan ?

The aim of the waste hierarchy is to extract the
maximum practical benefits from products and to
generate the minimum amount of waste.
Pengolahan berfungsi
• mengurangi potensi pencemaran dari
limbah yg meliputi fisik, biologi dan kimia
• Meningkatkan/menambah nilai guna dari
limbah
 Berbagai metode penanggulangan
limbah pencemar

• Secara fisik
• Secara kimiawi
• Secara biologi
– Bioteknologi pengolahan limbah pencemar
(Bioremediasi)
• Mikroba di alam secara umum berperan sebagai produsen, konsumen,
maupun redusen.
• Produsen menghasilkan bahan organik dari bahan anorganik dg energi
sinar matahari.
• Mikroba yg berperan sbg produsen adalah algae dan bakteri
fotosintetik.
• Konsumen menggunakan bahan organik yg dihasilkan oleh produsen.
• Contoh mikroba konsumen adalah protozoa.
• Redusen menguraikan bahan organik dan sisa-sisa jasad hidup yg mati
menjadi unsur-unsur kimia (mineralisasi bahan organik), sehingga di
alam terjadi siklus unsur-unsur kimia.
• Contoh bakteri redusen adalah bakteri dan jamur.
• Mikroba banyak dimanfaatkan di bidang lingkungan, terutama utk
mengatasi masalah pencemaran lingkungan (bioremidiasi), baik di
lingkungan tanah maupun perairan.
• Bahan pencemar :
• Bahan :
• Alami
• Sintetik,
• Sifat :
• Mudah dirombak (biodegradable),
• Sulit bahkan tdk bisa dirombak (rekalsitran/
nonbiodegradable)
• Beracun bagi jasad hidup dg bahan aktif tidak rusak dalam
waktu lama (persisten).
Air jernih, air yg keadaannya sudah kotor atau tercemar (sumur biasa,
sumur pompa, sumber mata-air, dsb.) di dalamnya ada bakteri, antara
lain :
•Kelompok bakteri besi (misalnya Crenothrix dan Sphaerotilus) yg
mampu mengoksidasi senyawa ferro menjadi ferri.
• air sering berubah warna kalau disimpan lama yaitu warna
kehitam-hitaman, kecoklat-coklatan, dsb.
•Kelompok bakteri belerang (mis: Chromatium dan Thiobacillus) yg
mampu mereduksi senyawa sulfat menjadi H2S.
• air disimpan lama akan tercium bau busuk (bau telur busuk).
•Kelompok mikroalge (termasuk mikroalga hijau, biru dan kersik)
• Air disimpan lama akan nampak jasad-jasad yg berwarna hijau,
biru atau pun kekuning-kuningan, tergantung kepada dominasi
jasad-jasad tersebut serta lingkungan yg mempengaruhinya.
Environmental Biotechnology
Environmental biotechnology is the solving of
environmental problems through the application of
biotechnology.
 Biotechnology and the Environment

❖Environmental Biotechnology -
the development, use and regulation of
biological systems for remediation of
contaminated environments (land, air,
water), and for environment-friendly
processes.
❖Bioremediation - the use of
microorganisms to remedy
environmental problems
 Why environmental biotechnology?
• It is needed to:
– eliminate the hazardous
wastes produced by our
other technologies.
– distinguish between
similar species and
ensure species are not at
risk of extinction.
– create alternative energy
sources (i.e. Biofuel).
 BIOREMEDIASI
• Bioremediasi merupakan suatu upaya pemulihan kondisi
(mengurangi polutan) dilingkungan dg menggunakan
aktivitas biologis (mikroorganisme) utk mendegradasi
dan/atau menurunkan toksisitas dari berbagai senyawa
pencemar.
• Mikroorganisme dari kelompok bakteri, khamir, dan
kapang merupakan kelompok utama yg berperan penting
dlm bioremediasi limbah pencemar di lingkungan
• Saat bioremediasi terjadi, enzim-enzim yg diproduksi oleh
mikroorganisme memodifikasi polutan dg mengubah struktur kimia
polutan tsbt, sebuah peristiwa yg dsbt biotransformasi.
• Biotransformasi berujung pada biodegradasi,

Pencemaran di lingkungan

Bioremediasi

Biodegradasi

Mikroorganisme Jenis substrat

Faktor lingkungan yg menjamin

Survival mikroba dan interaksinya

Monitoring
 Bioremediation treatment technologies
Treatments Definition
Bioaugmentation Addition of bacterial cultures to a
contaminated medium; frequently used in
bioreactors and ex situ systems
Biostimulation Stimulation of indigenous microbial
populations in soils and/ or ground water; may
be done in situ or ex situ
Bioreactors Biodegradation in a container or reactor; may
be used to treat liquids or slurries
Bioventing Methods of treating contaminated soils by
drawing oxygen through the soil to stimulate
microbial growth and activity
 Bioremediation treatment technologies

Treatments Definition

Composting Aerobic, thermophilic treatment

process in which contaminated
materials is mixed with a bulking
agent; can be done using static piles,
aerated piles, or continuously fed
reactor
 Bioremediasi didasarkan pada pemahaman atas tiga
prinsip utama:

(1) pengetahuan ttg fisiologi mikroba meliputi proses metabolisme

yg mengarah pd detoksifikasi senyawa berbahaya dan
pengendalian genetik mikroba yg mengontrol fungsi-fungsi tsbt

(2) pengetahuan ttg ekologi mikroba yg meliputi struktur dan

fungsi dari komunitas mikroba di alam;

(3) pengetahun ttg kerekayasaan (engineering) yaitu cara-cara

aplikasi proses-proses metabolisme mikroba di alam sesuai dg
yg diinginkan.
 Bioremediasi mempunyai dua tujuan yaitu

• menstimulasi pertumbuhan mikroba baik yg indigenus yaitu

mikroba asli maupun non indigenus atau mikroba yg sengaja
dimasukkan dari luar ke daerah yg terkontaminasi,
• menciptakan kondisi lingkungan yg sesuai utk meningkatkan
intensitas kontak langsung antara mikroba dg senyawa
kontaminan di lingkungan baik yg terlarut maupun yg terikat
oleh partikel utk mengalami biotransformasi, biodegradasi,
bahkan sampai biomineralisasi.
 Keberhasilan bioremediasi bergantung :

• karakteristik substrat pencemar yg akan didegradasi

oleh mikroba.
• keanekaragaman mikroba indigenus dan non
indigenus potensial pendegradasi limbah pencemar,
• faktor biotik dan abiotik lingkungan yg
mempengaruhi aktivitas biodegradasi limbah
pencemar
 Eksplorasi mikroba potensial

• Eksplorasi mikroba potensial yg dapat digunakan sbg

agen dlm bioremediasi umumnya dilakukan pada
lingkungan yg terkontaminasi oleh limbah pencemar
baik di tanah maupun di perairan.
• Kegiatan tersebut diarahkan selain utk mendapatkan
koleksi isolat mikroba indigenus juga utk mengurangi
kemungkinan terjadinya interaksi negatif ketika
dilakukan penambahan mikroba potensial tsb ke
suatu lingkungan tercemar.
 TEKNIK BIOREMEDIASI
• Bioremediasi diaplikasikan secara :
– langsung (in situ) pd lingkungan yg tercemar
• Sifat remediasinya secara alamiah (natural attenuation)
dan proses biodegradasi bahan pencemarnya
berlangsung sangat lambat
– Di luar lingkungan yg tercemar (ex situ), yaitu dg
membawa tanah/air yg terkontaminasi tsbt ke lokasi
pengolahan yg telah ditetapkan.
Tahapan bioremediasi tanah
Tahapan bioremediasi tanah
 Bioremediation Basics

❖ Aerobic and anaerobic

biodegradation
• Aerobic
• Oxygen is reduced to water and
the organic molecules (e.g.
petroleum, sugar) are oxidized
• Anaerobic
• An inorganic compound is reduced
and the organic molecules are
oxidized (e.g. nitrate is reduced
and sugar is oxidized)
• NOTE: Many microbes can do both
aerobic and anaerobic respiration; the
process which produces the most ATP
is used first!
 Bioremediation Basics
❖ The Players: Metabolizing Microbes
• Site usually contains a variety of microbes
• Closest to the contaminant: anaerobes
• Farthest away: aerobes
• The most common and effective bacteria are the indigenous
microbes (e.g. Pseudomonas in soil)
• Fungus and algae are also present in the environment and do
a good job of “cleaning up” chemicals (fungi do it better than
bacteria)
 Bioremediation Basics
❖ Bioremediation Genomics Programs
• Stimulating Bioremediation
• Add fertilizers (nutrient enrichment) to stimulate the
growth of indigenous microorganisms
• Adding bacteria or fungus to assist indigenous
microbes is known as bioaugumentation or seeding
 Bioremediation Basics
❖ Phytomediation
• Utilizing plants to clean up chemicals
• Ex: cottonwoods, poplar, juniper trees, grasses, alfalfa
• Low cost, low maintenance and it adds beauty to the site
 Cleanup Sites and Strategies
❖ Do the chemicals pose a fire or explosive hazard?
❖ Do the chemicals pose a threat to human health including the
health of clean-up workers? (what happened at Chernobyl to
the workers?)
❖ Was the chemical released into the environment through a
single incident or was there long-term leakage from a storage
container?
❖ Where did the contamination occur?
❖ Is the contaminated area at the surface of the soil? Below
ground? Does it affect water?
❖ How large is the contaminated area?
 Cleanup Sites and Strategies

❖ Soil Cleanup
• Either remove it (ex situ bioremediation) or in situ (in place)
• In place:
– If aerobic may require bioventing
– Most effective in sandy soils
• Removed:
– Slurry-phase, solid phase, composting, landfarming, biopiles
 Cleanup Sites and Strategies

❖ Bioremediation of Water
• Wastewater treatment
 Cleanup Sites and Strategies

❖ Bioremediation of Water
• Groundwater Cleanup
 Phytoremediation
• The word's etymology comes from the Greek φυτο (phyto) = plant,
and Latin « remedium » = restoring balance, or remediation.
• Phytoremediation describes the treatment of environmental
describes the treatment of environmental problems (bioremediation)
through the use of plants
• The use of plants to extract, sequester, detoxify, and/or
hyperaccumulate toxic pollutants from soil, water, and air.
• This process can be exponentiated through the application of
biotechnology and the expression of foreign genes by plants.
POTENSI BIOTEK PENGOLAH LIMBAH
1. Mendegradasi limbah spesifik di tempat dg menggunakan
biakan khusus atau gabungan
2. Inokulasi sistem pengolahan limbah konvensional dg
biakan khusus yg telah disesuaikan
3. Dekontaminasi dan detoksifikasi tumpahan
4. Peniadaan logam
5. Peniadaan secara biologis bau dan gas limbah yg
berbahaya seperti merkaptan, hidrogen, sulfida, sianida,
hidrogen berklor dsb
6. Menghasilkan biomassa dari limbah
7. Konversi limbah menjadi metan ( biogas)

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