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Pengantar Bioteknologi

FARMASI– UHAMKA
2014
Priyo Wahyudi
Pengantar Bioteknologi
• Definisi bioteknologi
• Ruang lingkup bioteknologi
• Sejarah bioteknologi
• State of the art bioteknologi
• Nilai strategis biotek (now & future).
Bioteknologi
• Dari akar kata:
– BIO = mahluk hidup, proses yang terjadi pada mahluk hidup
– TEKNOLOGI = kumpulan teknik untuk memecahkan masalah atau
membuat produk yang bermanfaat
• 3 Kata Kunci Bioteknologi:
1. Bios (materi hidup) yang meliputi : Virus, Bakteri, Fungi, Tanaman
dan Hewan
2. Sains dan teknologi, merupakan integrasi keduanya
3. Menghasilkan Produk atau Jasa, bagi kesejahteraan hidup manusia
• Bioteknologi merupakan pengintegrasian beberapa disiplin
ilmu pengetahuan alam dan teknologi : Mikrobiologi,
Fisiologi, Biokimia, Genetika, Biologi Molekuler, Kimia serta
rekayasa proses dan Teknik kimia.
Output Process Input
• Produk atau • Sains & • Bio
• Jasa • Teknologi
3 Kata Kunci Bioteknologi
Komponen “Bio”
• Organisme utuh : virus, sel bakteri, sel fungi, sel alga, sel
protozoa, individu tanaman, individu hewan, manusia.
• Bagian / komponen organisme : protein envelope, protein
capsid, protein core, dinding sel, flagela, pili, ribosom, DNA,
RNA
• Produk metabolit dari organisme : metabolit primer &
sekunder
• Agen biokonversi dari organisme : enzim , hormon,
imunoglobulin
• Bahan baku (raw material) yang berasal dari organisme :
tepung jagnng, tepung tapioka, cangkang kepiting, kulit
udang
Integrasi Ilmu & Teknologi dalam Bioteknologi

Crops Drugs Vaccines Diagnostics Livestock


Integrasi Ilmu & Teknologi
Perkembangan BIOTEKNOLOGI akan ditentukan kemajuan di
bidang:
SAINS: TEKNOLOGI :

BIOLOGI MOLEKULER TEKNIK KIMIA


GENETIKA TEKNIK MESIN
MIKROBIOLOGI TEKNIK ELEKTRO
FISIOLOGI TEKNIK INFORMATIKA
KIMIA
FARMASI
EKONOMI
MATERIAL
KOMPUTER
Output Bioteknologi
• Produk :
1. Makanan: keju, yoghurt, tempe, kecap, tauco, tape, oncom, tape, ketan
brem, arak, tuak, wine, beer, terasi, nata
2. Obat-obatan: antibiotik, hormon, vaksin, vitamin, antibodi monoklonal
3. Pertanian: pupuk bio, pestisida bio, kompos, bibit tanaman unggul
4. Peternakan: sapi daging, sapi susu, ayam daging, ayam telor, domba wol,
5. Fuel : bio etanol
6. Metabolit tanaman
7. Fragrance
8. Senjata biologis : toksin antrax, virus cacar
• Jasa :
1. Pengolahan limbah cair
2. Pengolahan limbah padat  kompos
3. Penanganan pencemaran minyak di laut
4. Forensik
5. Diagnostik
Definisi Bioteknologi
• Bioteknologi adalah integrasi ilmu pengetahuan alam dan
engineering untuk mendapatkan produk dan servis dari
organisme, sel, bagian darinya dan molekul-molekul
analognya (European Federation of Biotechnology, 1989)
• Bioteknologi adalah penggunaan organisme hidup atau teknik
biologis dalam industri yang dikembangkan melalui penelitian
dasar (Biotech Life Sciences Dictionary)
• Bioteknologi adalah suatu bidang penerapan biosains dan
teknologi yang menyangkut penerapan praktis organisme
hidup atau komponen subselulernya pada industri jasa dan
manufaktur serta pengelolaan lingkungan (Smith, 1993)
Ruang Lingkup Bioteknologi
• Bioteknologi telah menjadi bagian dari kehidupan sehari-hari
manusia, yang meliputi produk bahan pangan yang kita makan,
obat yang kita minum, dan tanaman yang kita tanam, serta
mengelola lingkungan kita.
• Sejak ditemukannya struktur DNA, lompatan besar telah dibuat
dalam memahami mekanisme fungsi sel, metabolisme, replikasi
dan pembentukan produk. Keuntungan ilmiah dari rekayasa
genetika dan biologi molekuler berlanjut dengan kecepatan
yang makin tinggi. Kemajuan ini membawa peluang dalam
menciptakan industri-industri baru yang didasarkan pada sel
hidup dan komponen sel.
• Bioteknologi digunakan dalam aktivitas yang sangat luas yang
mempunyai nilai sosial dan ekonomis yang penting
Ruang Lingkup Bioteknologi
1. Pertanian Agronomi, Perikanan, Peternakan
2. Pangan & Gizi
3. Farmasi
4. Medis & Kesehatan
5. Industri Kimia
6. Lingkungan
7. Militer
8. Material
9. Energi
Ruang Lingkup Bioteknologi
1. Pertanian Agronomi, Perikanan, Peternakan
– Pemuliaan tanaman untuk meningkatkan resistensi
terhadap hama, penyakit, kekeringan dan kondisi kadar
garam tinggi
– Perbanyakan massal dari bibit tanaman
– Pengembangan biopestisida
– Modifikasi tanaman untuk meningkatkan karakteristik
nutrisi dan proses
Marker
Methods for Plant Genetic Engineering are Well-Developed
and Similar to Those for Animals
Transgenic
Non-transgenic
Biological Insecticides
• Microbes that are
pathogenic to insects are
alternatives to chemical
pesticides in preventing
insect damage to
agricultural crops and
disease transmission
• Bacillus thuringiensis
infections are fatal in many
insects but harmless to
other animals, including
humans, and to plants
Roundup Transgenic Plant
Golden Rice is Modified to be Provide a Dietary Source of Vitamin A

Golden rice (yellow)


with standard rice
(white).

Worldwide, 7% of children suffer vitamin A deficiency, many of


them living in regions in which rice is a staple of the diet.
The Golden Rice Story
• Vitamin A deficiency is a major health problem
• Causes blindness
• Influences severity of diarrhea, measles
• >100 million children suffer from the problem

• For many countries, the infrastructure doesn’t exist


to deliver vitamin pills

• Improved vitamin A content in widely consumed crops


an attractive alternative
-Carotene Pathway in Plants
IPP

Geranylgeranyl diphosphate
Phytoene synthase

Phytoene
Problem: Phytoene desaturase
Rice lacks
these enzymes ξ-carotene desaturase

Lycopene
Lycopene-beta-cyclase
Normal
Vitamin A  -carotene
“Deficient”
Rice (vitamin A precursor)
The Golden Rice Solution
-Carotene Pathway Genes Added
IPP

Geranylgeranyl diphosphate
Daffodil gene Phytoene synthase

Phytoene
Vitamin A Phytoene desaturase
Pathway Single bacterial gene;
performs both functions
is complete ξ-carotene desaturase
and functional Lycopene
Daffodil gene Lycopene-beta-cyclase

Golden  -carotene
Rice (vitamin A precursor)
Ruang Lingkup Bioteknologi
• Perikanan & Peternakan
‒ Produksi vaksin
‒ Pengendalian fertilitas
‒ Persilangan hewan ternak
‒ Kloning
Pembenihan Tuna

Ikan salmon hasil rekayasa genetik (“biotech”) salmon (left)


dan standard salmon (right).
Ruang Lingkup Bioteknologi
2. Pangan & Gizi
– Produksi roti, keju, yoghurt dan pangan fermentasi seperti
cuka dan kecap
– Pembuatan keju, tempe dan sosis
– Pembuatan bir dan wine
– Produksi flavour dan bahan pewarna
Ruang Lingkup Bioteknologi
3. Farmasi
– Pengembangan molekul baru untuk pengobatan (novel
therapeutic molecules for medical treatments)
– Pembuatan diagnostic kits
– Sistem delivery obat
Penicillin fermentation
Recombinant DNA Tech – Human Insulin
Production by Bacteria
Human Insulin Production by Bacteria

and cut with a restriction enzyme

6) join the plasmid and human fragment


Human Insulin Production by Bacteria

Mix the recombinant plasmid with


bacteria.

Screening bacterial cells to learn which contain the human insulin


gene is the hard part.
Route to the Production by Bacteria of Human Insulin
One cell with the
recombinant plasmid

A fermentor used to grow


recombinant bacteria.

This is the step when gene cloning takes place.


The single recombinant plasmid replicates within a cell.
Then the single cell with many recombinant plasmids produces
trillions of like cells with recombinant plasmid – and the human
insulin gene.
Route to the Production by Bacteria of Human Insulin

The final steps are to collect the bacteria, break open the cells, and
purify the insulin protein expressed from the recombinant human
insulin gene.
Rapid test

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Test device component categories
Porous materials
Sample pads
Conjugate pads
Membranes Absorbent pads

Reagents
Capture antibodies and/or antigens (test line and control line)
Conjugate ligand (antibody or antigen)
Detector particle (e.g., colloidal gold)
Blocking agents, detergents, surfactants, stabilizers, buffers, etc.

Housing and lamination materials


Back laminate (for holding porous components together)
Top laminate (optional, to act as a "splash guard" or prevent evaporation and back-
migration of detector reagent)

Device housing (optional—if used, the housing typically comprises two, snap-fit
plastic pieces that envelop the reagent-loaded porous media assembly and facilitate
sample addition)

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Rapid immunoassay

40
Rapid immunoassay

41
42
Rapid lateral flow test

43
Pregnancy test

44
Specifications for a pregnancy test
Following are the development guidelines given to the research and development manager responsible for
designing, developing, and scaling up to manufacturing a hypothetical lateral-flow diagnostic test device for
detecting human chorionic gonadotropin (hCG) in urine.
The hCG hormone is measured in milli-international units per milliliter (mIU/ml).

• More than 99% of specimens containing 25 mlU/ml will produce a visible signal at the test line.

• More than 99% of specimens containing 5 mlU/mL will not produce a visible signal at the test line.

• The test will have a control line that will always produce a visible signal if the test is performed correctly and
if all of the test components are functional.

• If the sample is positive (contains 25 mIU/ml hCG), the test must produce a visible signal at the test line
within 3 minutes of sample addition.

• The test result must be stable (not change) for at least 30 minutes after the sample is added.

• The test device will be contained in a plastic housing and designed to be compatible with "in-stream"
sampling protocols.

• The packaged (unused) product must be stable for at least 18 months at ambient conditions (15°–30°C).

• Total incremental manufacturing cost per test (excluding licensing and royalties) can not exceed $0.25 US.

45
Rapid test for drugs of abuse

46
47
Ruang Lingkup Bioteknologi
4. Medis & Kesehatan
– Rekayasa jaringan untuk mengganti organ
– Stem cell
– Terapi gen
Terapi Gen
Terapi Gen

• Andrew Gobea was diagnosed with ADA (Adenosin Deaminase) deficiency


before he was born. Without this enzyme, the body is unable to break down a
toxic substance called deoxyadenosine. The toxin builds up and destroys
infection-fighting immune cells called T and B lymphocytes.
• This lack of the ADA enzyme causes an immune deficiency disease called
SCID (Severe Combined immunodeficiency).
• Andrew was given the gene for a functional ADA enzyme four days after his
birth in 1993.
• Umbilical cord blood was collected at birth.
• Stem cells were isolated and mixed with a virus carrying a functional ADA
gene.
• Stem cells were returned to Andrew with the aim of populating his bone
marrow with cells that could make the ADA enzyme.
Two Years Later

• Andrew has been maintained on costly injections of purified


ADA enzyme while waiting to see if the gene therapy has been
effective.
• Tests have shown that the functional ADA gene was introduced
into 0.01% of Andrew’s stem cells and this population has given
rise to 5-7% of his white blood cells. Is this enough for a cure?
Four Years Later
• Andrew’s physician decides to taper off the enzyme injections to
see if Andrew’s immune system can protect him.
Ruang Lingkup Bioteknologi
5. Industri Kimia
– Produksi bahan kimia dan pelarut seperti etanol, asam
sitrat, aseton dan butanol
– Sintesis bahan kimia adi seperti enzim, asam amino, dan
alkaloid
Ruang Lingkup Bioteknologi
6. Lingkungan
– Recovery biologis logam-logam berat dari limbah
pertambangan dan industri
– Bioremediasi tanah dan air yang tercemar bahan kimia
beracun
– Pengolahan sampah dan air limbah
– Bioleaching
– Biosensors: mendeteksi senyawa toksik di lingkungan,
contoh: RIS® Water tests yi antibody based kit to detect low
level of solvents such as benzene
A. ferrooxidans and its proposed
role in bioleaching. The
chemolithoautotrophic metabolism
of A. ferrooxidans results in the
oxidation/reduction of iron and sulfur
compounds and the solubilization of
copper and other commercially
valuable metals in a process called
bioleaching or biomining. It also
results in the production of acidified
solutions in pristine environments and
acid mine drainage in bioleaching
operations. A) Model of copper
bioleaching by A. ferrooxidans. B)
Oxidation/reduction reactions carried
out by A. ferrooxidans. The scheme
provided here presents the basic
concepts of bioleaching and further
details are provided in the review [4].
C) Acid mine drainage in the Rio
Tinto, Spain, derived from naturally
occurring pyritic ore bodies and
abandoned mine workings initiated in
pre-Roman times [3]. D) Commercial
bioleaching heap for copper recovery,
Chile. 3PG: 3-phosphoglycerate.
Ruang Lingkup Bioteknologi
7. Militer
– Senjata biokimia dan biologis
– Biosensor anti toksin
– Anti Bioterorisme
– Forensik DNA
Applications of Biotechnology
for Identification: Forensics

1 2 3 C  S 4 5 67
R  C
I  E
Suspects M  N Suspects
E  E
Applications of Biotechnology
for Identification: Paternity Testing

X X

X X

X X

X X
Ruang Lingkup Bioteknologi
8. Material
– Biomaterial untuk operasi dan pembedahan
– Biomaterial untuk otomotif
– Biomaterial untuk akustik

Jaring laba-laba 5x lebih kuat dari


Artificial epidermis untuk operasi
baja dengan diameter yang sama
Bioselulosa (nata) dijadikan sebagai membran pada speaker
Ruang Lingkup Bioteknologi
9. Energi
– Biofuel Ethanol
– Biodiesel
– Biohidrogen fuel cell
5. Bio-fuels

• Plant-derived fuels: plant


species for hydrocarbon (oil)
production, e.g. rape-seed,
sunflower, olive, peanut oils. Or
ethanol production of sugars (or
cellulose) derived from plants.
• Conversion of used cooking oil to
bio-fuel (called bio-diesel)
• Biogas: gases from composts or
landfill, but methane is a green
house gas
Bioethanol and biofuel cell:
• Sugar cane, sugar beet wastes, high starch material (cassava,
potatoes, millet) to be hydrolyzed by starch hydrolyzing
enzyme to convert sucrose or glucose to ethanol. Mainly used
in Brazil.
• Corn ethanol: 22% less carbon emission, used in the US.
• Bio-diesel: 68% less carbon emission; oils from soybean (US) or
canola oil (Germany)
• Cellulosic ethanol: 91% less carbon emission, but difficult to
change cellulose to ethanol
• Hydrogen energy however is the trend of future renewable
energy without carbon emission: a journey to forever…….
• Problem is how to generate the hydrogen; too costly with
conventional chemical methods or reverse osmosis.
A Journey forever?
• Various bacteria and algae, for example Escherichia coli,
Enterobacter aerogenes, Clostridium butyricum, Clostridium
acetobutylicum, and Clostridium perfringens have been
found to be active in hydrogen production under anaerobic
conditions.
• The most effective H2 production is observed upon
fermentation of glucose in the presence of Clostridium
butyricum (strain IFO 3847, 35 mmol h–1 H2 evolution by 1
g of the microorganism at 37°C).
Fuel ethanol
Hydrogen Fuel Cells
• Fuel cells are similar to
batteries, but designed for
continuous replenishment
of energy via external fuel
• Many different types of fuel
cells, most common will
likely be the PEM FC
• Hydrogen and oxygen in,
water vapor and liquid
water out
• Typical output is about .8
volts
science of hydrogen fuel cells
A Pathway for our Future Energy?
A microbial
biofuel cell:
(A) With a microbial
bioreactor providing
fuel separated from the
anodic compartment of
the electrochemical cell.

(B) With a microbial bioreactor


providing fuel directly in the anodic
compartment of the electrochemical
cell.
The Working principle Of An Enzyme Fuel Cell

The enzyme and mediator are immobilized on


the anode.

Rough layout of the anode structure


General characteristics of chemical and biological fuel cell

Chemical Fuel Cell Biological Fuel Cell


Catalyst Noble Metals Microorganism /
enzyme
pH Acidic Solution Neutral Solution pH
(pH<1) 7.0-9.0
Temperature over 200 ° C Room Temperature
22-25 ° C
Electrolyte Phosphoric-acid Phosphate Solution
Capacity High Low
Efficiency 40 - 60 % over 40 %
Fuel Type Natural gas, H2, etc. Any Carbohydrates
and hydrocarbons
*conversion rate 50%
Theoretical energy content of methanol, ethanol, and glucose.
The calculation is based on the assumption of complete
conversion; the likely conversion rate in practice is around 50 %.

Methanol Ethanol Glucose


(1 g/10 ml H2O) (1 g/10 ml H2O) (3 g /10 ml H2O)

Capacity [m Ah] 5025 (2512*) 3500 (1750*) 893 (446*)


H2O formed [g] 1.125 0.391 0.300
CO2 formed [g] 1.375 0.956 2.97 glucono-
lactone
Fuel Cell Parts –
Form and Function
The Energy Problem
• How will society meet
growing energy
demands in a
sustainable manner?
• Fossil-fuels currently
supply ~80% of world
energy demand.
Are Biofuels the Answer?...
Biofuels as an Alternative
• Biofuels are not THE answer to sustainable
energy, but biofuels may be part of the answer
• Biofuels may offer advantages over fossil fuels,
but the magnitude of these advantages
depends on how a biofuel crop is grown and
converted into a usable fuel
Analysis of Alternative Biofuels
• “First generation” biofuels: food-based
biofuels that are currently commercially
available:
– Corn-grain ethanol
– Soy Biodiesel
• “Second generation” biofuels: cellulosic
biofuels of the future
– Diverse prairie biomass
What is Biodiesel?
• Alternative fuel for diesel engines
• Made by chemically combining any natural oil or fat (from
vegetable oil or animal fat) with an alcohol
• Lower emissions, High flash point (>300F), Safer
• Biodegradable, Essentially non-toxic.
• Chemically, biodiesel molecules are mono-alkyl esters
produced usually from triglyceride esters
Fatty Acid
FA Alcohol
Glycerin

FA FA

FA Biodiesel
Vegetable Oil
Bio diesel Process Flow Diagram

NRRaje Feb 06
Biodiesel Samples
NRRaje Feb 06
Milestone Bioteknologi
FERMENTASI
· Proses kimiawi dari senyawa organik yang dimediasi oleh enzim.
· Fermentasi dikenal sejak 6000 SM (Sumeria dan Babilonia membuat bir)
· Bioteknologi tradisional à Bioteknologi moderen
· Disain Bioreaktor
· Bidang: Pangan, produksi biomassa, antibiotika, enzim dll.

REKAYASA GENETIKA
· Upaya merekayasa materi genetik organisme untuk meningkatkan
kemampuannya
· RG pada mikroorganisme à industri antibiotika, enzim dan asam amino, dll
· RG pada tanaman à tanaman transgenik antihama (Bt cotton / Bt corn),
tanaman tahan kondisi lingkungan ekstrim (tahan alkali, tahan garam, tahan
logam berat, tahan kekeringan), tanaman super dengan produksi yang
berlipat ganda, dll
· RG pada hewan spt. ulat sutera, hama infertil, domba wool, dll
· Kloning
Milestone Bioteknologi
TEKNOLOGI ENZIM
· Enzim diproduksi oleh organisme (mikroorganisme, hewan dan tumbuhan)
diaplikasikan di industri makanan, minuman, manufaktur, dll
· Kompetensi enzim adalah mengurangi energi yang diperlukan untuk terjadinya
proses biokonversi
· Perkembangan selanjutnya à enzim mikroorganisme mendominasi karena segi
kemudahan produksinya
· Enzim protease, karbohidrase, lipase, lainnya

REKAYASA BIOKIMIA
· Proses biokimiawi dalam metabolisme mikroorganisme dapat direkayasa à
Biokonversi / Biotransformasi

REKAYASA PRODUK / SISTEM


· Produk molekul biologi seperti antibodi atau enzim dapat dikembangkan
sebagai BIOSENSOR untuk tujuan diagnosis dan detoksifikasi
· Perkembangan genetika molekuler memungkinkan pengobatan suatu penyakit
herediter ataupun degeneratif melalui terapi gen
Timeline Bioteknologi
Timeline Bioteknologi
• Awal mula manusia mengenal Bioteknologi dimulai sejak 10.000
tahun lalu pada awal peradaban pertanian (early agrarian
societies) dimana orang mengumpulkan benih tanaman dengan
sifat yang dikehendaki untuk ditanam pada tahun berikutnya
dan melakukan persilangan yang selektif untuk meningkatkan
kualitas hewan ternak, seperti di Babylonia, Mesir dan Romawi.
• Pada 6000 SM orang telah membuat bir, anggur dan roti melalui
fermentasi, sebuah proses alami dimana aktifitas organisme
bersel tunggal memainkan peran yang penting.
• Pada 4000 SM di Cina telah menggunakan bakteri asam laktat
(BAL) untuk membuat yogurt, kapang untuk membuat keju dan
bakteri asam asetat untuk membuat cuka.
Fermentasi Mesir Kuno
Timeline Bioteknologi
• Pada tahun 1500 sesudah masehi, ekspedisi pengumpulan
tanaman telah mencakup seluruh penjuru dunia. Koleksi
tersebut dapat membuat Bank Gen Tanaman yang pertama.
• Nikolai I. Vavilov (1887 – 1943) ahli genetika
tanaman dan agronomi Russia menginisiasi
program riset dan persilangan yang komprehensif
yang mencakup suatu rencana logis untuk
manajemen sumber genetika tanaman. Ia
mengkoleksi hampir seluruh jenis tanaman di
dunia yang kemudian mengantarnya pada teori "
asal-usul penyebaran" tanaman budidaya. Selain
itu, ia juga mempelajari cara-cara memperbaiki
hasil jagung, gandum, dan serealia lainnya untuk
mendukung pertumbuhan penduduk dunia.
Timeline Bioteknologi

Akhir abad 19 merupakan sebuah tonggak (milestone)


dalam biologi. Gregor Mendel (1822 - 1884) memulai
studi genetika menggunakan biji dan tanaman.
Timeline Bioteknologi
• Istilah "BIOTEKNOLOGI" dimunculkan pertama kali pada
tahun 1917 oleh Karl Ereky (1878 – 1952), seorang
insinyur Hungaria.
– Produksi daging babi menggunakan gula bit sebagai pakan
– BIOTEKNOLOGI didefinisikan sebagai semua lini pekerjaan yang
dihasilkan dari raw material dengan bantuan organisme hidup.

• Bioteknologi diterapkan di industri


dan pertanian bersamaan pada
awal abad ke-20. Proses fermentasi
dikembangkan diantaranya untuk
memproduksi aseton dari pati dan
cairan cat untuk mobil.
Timeline Bioteknologi
• Antara tahun 1930 - 1952,
peneliti memfokuskan pada
hubungan antara gen dan
protein. Mereka menemukan
suatu hubungan langsung
antara mutasi sebuah gen
dan sekuens asam amino
dari sebuah protein.
• Penemuan antibiotik
penisilin oleh Alexander
Fleming (1928) telah
membawa perkembangan
biteknologi ke arah farmasi.
A Fortunate Accident—Antibiotics

• 1928: Alexander Fleming


discovered the first
antibiotic
• Fleming observed that
Penicillium fungus made
an antibiotic, penicillin,
that killed S. aureus
• 1940s: Penicillin was
tested clinically and mass
produced
Timeline Bioteknologi

• Pada tahun 1961 Bioteknologi diredifinisi sebagai


industrialisasi produksi barang atau jasa melalui suatu
sistem dan proses biologis pada suatu organisme

Bioengineered Biotechnology Process


Bioengineered Biotechnology Process

• Upstream processing
 Preparation of raw material
• Fermentation and
biotransformation
 Use of bioreactor
• Downstream processing
 Recovery and purification
from the medium or cell
mass
Timeline Biotechnology
• In the 1960’s and 1970’s focused primarily on improving
upstream processing, bioreactor design and downstream
processing
• New strains created by mutation and screening
• Very successful for antibiotic production
• Everything changes with development of recombinant DNA
technology
• New organisms for production (transfer production to new species)
• Quantity production of molecules normally present at miniscule levels
Timeline Bioteknologi
1950 to 1960
• 1951 McClintock discovers transposable elements, or "jumping genes," in corn.
• 1953 Watson and Crick reveal the three-dimensional structure of DNA.
• 1954 Cell-culturing techniques are developed.
• 1955 An enzyme involved in the synthesis of a nucleic acid is isolated for the first
time.
• 1956 The fermentation process is perfected in Japan.
Kornberg discovers the enzyme DNA polymerase I, leading to an understanding
of how DNA is replicated.
• 1957 Sickle cell anemia is shown to occur due to a change of a single amino acid.
• 1960 Exploiting base pairing, hybrid DNA-RNA molecules are created.
Messenger RNA is discovered.
• 1961 The genetic code is understood for the first time.
• 1964 The existence of reverse transcriptase (RT) is predicted.
• 1967 The first automatic protein sequencer is perfected.
• 1969 An enzyme is synthesized in vitro for the first time.
Timeline Bioteknologi
1970 Specific restriction nucleases are identified, opening the way for gene cloning. RT is
discovered independently in murine and avian retroviruses.
1971 RT is shown to have ribonuclease H (Rnase H) activity.
1972 The DNA composition of humans is discovered to be 99 percent similar to that of
chimpanzees and gorillas. Purified RT is first used to synthesize cDNA from purified mRNA in
vitro.
1973 Cohen and Boyer perform the first successful recombinant DNA experiment, using
bacterial genes.
1974 The National Institute of Health forms a Recombinant DNA Advisory Committee
to oversee recombinant genetic research.
1975 Colony hybridization and Southern blotting are developed for detecting specific DNA
sequences. The first monoclonal antibodies are produced.
1976 The tools of recombinant DNA are first applied to a human inherited disorder.
Molecular hybridization is used for the prenatal diagnosis of alpha thalassemia.
Yeast genes are expressed in E. coli bacteria.
1977 Genetically engineered bacteria are used to synthesize human growth protein.
1978 North Carolina scientists Hutchinson and Edgell show it is possible to introduce specific
mutations at specific sites in a DNA molecule.
1979 The first monoclonal antibodies are produced.
Timeline Bioteknologi
1980 The U.S. patent for gene cloning is awarded to Cohen and Boyer.
1981 The first gene-synthesizing machines are developed.
The first genetically engineered plant is reported.
Mice are successfully cloned.
1982 Humulin, Genentech's human insulin drug produced by genetically engineered
bacteria for the treatment of diabetes, is the first biotech drug to be approved by
the FDA.
1983 The Polymerase Chain Reaction (PCR) technique is conceived.
The first genetic transformation of plant cells by TI plasmids is performed.
The first artificial chromosome is synthesized.
The first genetic markers for specific inherited diseases are found.
Efficient methods are developed to synthesize double-stranded DNA from first-
strand cDNA involving minimal loss of sequence information.
1984 The DNA fingerprinting technique is developed.
The first genetically engineered vaccine is developed.
Chiron clones and sequences the entire genome of the HIV virus.
1985 Fully active murine RT is cloned and overexpressed in E. coli.
Timeline Bioteknologi
1986 The first field tests of genetically engineered plants (tobacco) are conducted.
Ortho Biotech's Orthoclone OKT3, used to fight kidney transplant rejection, is
approved as the first monoclonal antibody treatment.
The first biotech-derived interferon drugs for the treatment of cancer, Biogen's
Intron A and Genentech's Roferon A, are approved by the FDA.
The first genetically engineered human vaccine, Chiron's Recombivax HB, is
approved for the prevention of hepatitis B.
1987 Humatrope is developed for treating human growth hormone deficiency.
Genentech's tissue plasminogen activator (tPA), sold as Activase, is approved
as a treatment for heart attacks.
1988 Congress funds the Human Genome Project, a massive effort to map and
sequence the human genetic code as well as the genomes of other species.
1989 Amgen's Epogen is approved for the treatment of renal disease anemia.
Microorganisms are used to clean up the Exxon Valdez oil spill.
The gene responsible for cystic fibrosis is discovered.
Timeline Bioteknologi
1990 The first approved gene therapy treatment for immune disorder.
1991 Amgen develops Neupogen, for the treatment of low white blood cells in
chemotherapy patients.
Immunex's Leukine, used to replenish white blood counts after bone marrow
transplants, is approved.
Genzyme's Ceredase is approved for the treatment of Gaucher's disease.
1992 The three-dimensional structure of HIV RT is elucidated.
Recombinate, used in the treatment of hemophilia A
Chiron's Proleukin is approved for the treatment of renal cell cancer.
1993 Chiron's Betaseron is approved as the first treatment for multiple sclerosis .
The FDA declares that genetically engineered foods are "not inherently
dangerous" and do not require special regulation.
1994 Genentech's Nutropin is approved for the treatment of growth hormone
deficiency.
The first breast cancer gene is discovered.
1995 The first baboon-to-human bone marrow transplant is performed on an AIDS
patient.
Timeline Bioteknologi
1996 Biogen's Avonex is approved for the treatment of multiple sclerosis.
Scottish scientists clone identical lambs from early embryonic sheep.
1997 Scottish scientists report cloning a sheep, using DNA from adult sheep cells.
A group of Oregon researchers claims to have cloned two Rhesus monkeys.
A new DNA technique combines PCR, DNA chips, and a computer program,
providing a new tool in the search for disease-causing genes.
1998 University of Hawaii scientists clone three generations of mice from nuclei of
adult ovarian cumulus cells.
Human skin is produced in vitro.
Embryonic stem cells are used to regenerate tissue and create disorders
mimicking diseases.
A rough draft of the human genome map is produced, showing the locations of
more than 30,000 genes.
1999 The complete genetic code of the human chromosome is first deciphered.
The rising tide of public opinion in Europe brings biotech food into the
spotlight.
Timeline Bioteknologi
2000 A rough draft of the human genome is completed by Celera Genomics and
the Human Genome Project.
Pigs are the next animal cloned by researchers, hopefully to help produce
organs for human transplant.
"Golden Rice," modified to make vitamin A, promises to help third-world
countries alleviate blindness.
2001 The sequence of the human genome is published in Science and Nature,
making it possible for researchers all over the world to begin developing
treatments.
2002 Scientists complete the draft sequence of the most important pathogen of
rice, a fungus that destroys enough rice to feed 60 million people annually. By
combining an understanding of the genomes of the fungus and rice, scientists
will elucidate the molecular basis of the interactions between the plant and
pathogen.
2003 Dolly, the cloned sheep that made headlines in 1997, is euthanized after
developing progressive lung disease. Dolly was the first successful clone of a
mammal.
Tingkatan Bioteknologi
State of the Art
1. LOW BIOTECHNOLOGY
· Volume besar dengan produk atau jasa bernilai rendah (Low value high volume)
· Investasi murah, skala operasi kecil – menengah, labour intensive, sistem nonsteril,
teknologi pedesaan / home industry
· Pemurnian air, pengolahan limbah cair dan sampah, gas metan, etanol, biomas (PST)
atau pakan ternak

2. MIDLE BIOTECHNOLOGY
· Volume relatif besar dengan produk bernilai ekonomi menengah (Midle value and
volume)
· Investasi menengah, sistem operasi tidak terlalu rumit
· Makanan minuman fermentasi, pakan ternak, biofertilizer, biopestisida, crude enzyme

3. HIGH BIOTECHNOLOGY
· Volume kecil dengan nilai ekonomis yang tinggi pula (High value low volume)
· Investasi tinggi, pabrik dan sistem operasi rumit, terkontrol
· Antibiotika, vaksin, antibodi, enzim dan vitamin
Nilai Strategis Bioteknologi

1. Pertambahan populasi manusia yang tidak dapat


diimbangi dengan pertambahan produksi bahan
kebutuhan hidup yang dihasilkan secara konvensional.
2. Menipisnya sumber alam yang tidak dapat diperbaharui
3. Keberlimpahan keanekaragaman sumber gen di alam
yang belum dimanfaatkan secara maksimal oleh
manusia.
4. Perkembangan IPTEK manusia yang mengarah pada
peningkatan efektivitas dan efisiensi serta nilai
ekonomis
IKHTISAR SKEMATIS PROSES BIOTEKNOLOGI

SELEKSI
ORGANISME

GENETIKA TERAPAN
Mutasi, rekombinasi, manipulasi gen, transformasi
gen, kloning

Udara Energi

PRODUK
BAHAN BAKU BIOREAKTOR
Atau
Seleksi, persiapan, Mikrobial, sel hewan,
perlakuan sel tanaman, enzim JASA
Sterilisasi Dowstream
process
Heat atau
FORMULASI
Recovery
PENGENDALIAN
PROSES
Biotech: Present & Future
Transfer of new Anti-cancer drugs
Culture of plants
genes into animal from single cells Diagnostics
organisms

Cell Monoclonal
Culture Antibodies
Crime solving
Molecular
Biology

DNA Tracers
technology Genetic
Engineering
Synthesis of
Banks of Cloning specific DNA
DNA, RNA Synthesis
probes
and proteins of new Mass prodn. of
proteins human proteins
Complete Localisation of
New types of Resource bank
map of the genetic disorders
plants and for rare human
human
animals chemicals
genome
New
New types antibiotics
of food Gene therapy
Fakta Industri Bioteknologi
• Biotechnology is a $30 billion a year industry that has produced some
160 drugs and vaccines.
• There are more than 370 biotech drug products and vaccines
currently in clinical trials targeting more than 200 diseases, including
various cancers, Alzheimer’s disease, heart disease, diabetes, multiple
sclerosis, AIDS and arthritis.
• Biotechnology is responsible for hundreds of medical diagnostic tests
that keep the blood supply safe from the AIDS virus and detect other
conditions early enough to be successfully treated. Home pregnancy
tests are also biotechnology diagnostic products.
• Genetic engineering is sweeping the world’s farms. Seven million
farmers in 18 countries grew genetically engineered crops on 16.72
million acres last year.
• Consumers already are enjoying biotechnology foods such as papaya,
soybeans and corn. Hundreds of biopesticides and other agricultural
products also are being used to improve our food supply and to
reduce our dependence on conventional chemical pesticides.
Fakta Industri Bioteknologi
• Environmental biotechnology products make it possible to clean up
hazardous waste more efficiently by harnessing pollution-eating
microbes without the use of caustic chemicals.
• Industrial biotechnology applications have led to cleaner processes
that produce less waste and use less energy and water in such
industrial sectors as chemicals, pulp and paper, textiles, food, energy,
and metals and minerals. For example, most laundry detergents
produced in the United States contain biotechnology-based enzymes.
• DNA fingerprinting, a biotech process, has dramatically improved
criminal investigation and forensic medicine, as well as afforded
significant advances in anthropology and wildlife management.
• There are 1,473 biotechnology companies in the United States, of
which 314 are publicly held.
• Market capitalization, the total value of publicly traded biotech
companies (U.S.) at market prices, was $311 billion as of mid-March
2004.
Fakta Industri Bioteknologi
• The biotechnology industry has mushroomed since 1992, with U.S.
revenues increasing from $8 billion in 1992 to $39.2 billion in 2003.
• The U.S. biotechnology industry employed 198,300 people as of Dec.
31, 2003.
• Biotechnology is one of the most research-intensive industries in the
world. The U.S. biotech industry spent $17.9 billion on research and
development in 2003.
• The top eight biotech companies spent an average of $104,000 per
employee on R&D in 2003.
• The biotech industry is regulated by the U.S. Food and Drug
Administration (FDA), the Environmental Protection Agency (EPA) and
the Department of Agriculture (USDA)
Designer Jeans: Made by Microbes?

• Stone-washing: Trichoderma
• Cotton: Gluconacetobacter
• Debleaching: Mushroom peroxidase
• Indigo: E. coli
• Plastic: Bacterial polyhydroxyalkanoate
Stone washing denim
Indigo dye adheres
to denim surface
traditional new method
method

Cellulase enzyme
Denim is faded by removes some of the dye
abrasive action of by partially hydrolyzing the
pumice stones cotton surface

• new looks
• weakens the fabric • lower costs
• shorter treatment times
• less solid waste
Detergents
• Detergent industry is the largest single market for enzymes at 25
- 30% of total sales
• Dirt comes in many forms and includes proteins, starches and
lipids (fats and oils)
• proteases, amylases, lipases are enzymes used in detergents
• enzymes allows lower temperatures and less agitation for
washing

Inner core of enzyme plus


preservative bound with CMC

Protective waxy coat that


disperses in the wash
Industrial Chemicals
Examples:
• organic acids produced from Aspergillus niger,
citric acid used in soft drinks
• Xylanase used for wood pulping and bleaching
Agricultural
Examples:
• Recombinant bovine somatotropin (bST) for
increasing milk production
• Bio-insecticides for crop protection
• Phyto-vanilla(tm) flavor derived from tissue
culture
The Birth of Modern Chemotherapy
• Treatment with chemicals is chemotherapy
• Chemotherapeutic agents used to treat infectious disease can
be synthetic drugs or antibiotics
• Antibiotics are chemicals produced by bacteria and fungi that
inhibit or kill other microbes
The First Synthetic Drugs
• Quinine from tree bark was long used to treat malaria
• Paul Erlich speculated about a “magic bullet” that could
destroy a pathogen without harming the host
• 1910: Ehrlich developed a synthetic arsenic drug, salvarsan, to
treat syphilis
• 1930s: Sulfonamides were synthesized
Microbial Ecology
• Bacteria recycle carbon, nutrients, sulfur, and phosphorus that
can be used by plants and animals
Biotechnology
• Recombinant DNA technology, a new technique for
biotechnology, enables bacteria and fungi to produce a variety
of proteins including vaccines and enzymes
– Missing or defective genes in human cells can be replaced in gene
therapy
– Genetically modified bacteria are used to protect crops from insects
and from freezing
• Mutant strains of bacteria and fungi that
synthesize large amounts of metabolites
• Primary metabolites - produced during major
metabolic pathways and are essential to
microbe’s function – amino acids, organic acids
synthesized during logarithmic growth
• Secondary metabolites – by-products of
metabolism that may not be critical to microbe’s
function – vitamins, antibiotics, and steroids
synthesized during stationary phase
123
Insert figure 26.35
Origins of metabolites

124
• Many syntheses occur in sequential fashion
involving more than one organism.
• Biotransformation – waste product of one
organism becomes the building block of the
next

125
Insert figure 26.36
biotransformation

126