ENZIM

 Proses Konversi Pada Bahan yang Memanfaatkan

Enzim sebagai Katalis
 Enzim

• Molekul protein yang dihasilkan sel hidup  katalis reaksi

biokimia  spesifik & selektif

• Secara molekuler:

 merupakan protein yang tersusun atas serangkaian

asam amino dalam komposisi dan sekuens yang
teratur dan tetap  konformasi 3 dimensi

 “Sisi aktif ” = bagian permukaan enzim yang berupa

rangkaian asam amino  gugus fungsionalnya
berinteraksi dengan substrat
Enzyme Structure

Enzymes are globular proteins. Their folded conformation

creates an area known as the active site. The nature and
arrangement of amino acids in the active site make it
specific for only one type of substrate.

http://science.howstuffworks.com/environmental/life/cellular-microscopic/cell2.htm
 Reaksi Enzimatis

 Kerja enzim: mempercepat reaksi dengan menurunkan

energi aktivasi (Ea)
Pengaruh Enzim pada Energi Aktivasi
Aktivitas Enzim
 Aktivitas enzim dinyatakan sebagai ‘unit’

 Definisi:
‘Satu unit merupakan sejumlah enzim yang akan
mengkonversi satu mikro mol substrat per menit pada
kondisi pH dan suhu tertentu’.

• Aktivitas Enzim  μ/mL

• Aktivitas Spesifik  μ /mg protein enzim


ukuran kemurnian
 Laju. reaksi Laju maksimum Laju maksimum

Laju reaksi
pH optimum suhu optimum

pH suhu
a. Pengaruh pH b. Pengaruh suhu

Enzim1 aktivator
Laju. reaksi

Pembentu
Laju maksimum
Vm

Produk
enzim

kan
V m
2 Km
Waktu
[substrat] d. Pengaruh aktivator
Vm: Laju Reaksi maksimum
Km: konstanta Michaelis enzim
Pembentu

c. Pengaruh [substrat] Produk

Enzim1 inhibitor
kan

Waktu
d. Pengaruh inhibitor
Pengaruh Substrat

K1 K2
E+S K--1
ES E+P

Vmaks S 
 V 
K m  S   Persamaan Michaelis-Menten

Saat V = ½ Vmaks  Km = [S]

Vmaks

½ Vmaks

Km →[S]
Line Weaver-Burk  1  K m . 1  1
V V maks S  V maks

1 a b
V Km
Slope 
Vmaks
1
Vmaks
1
1 S 

Km

Km  memberikan informasi tentang ikatan substrat yang

terikat oleh enzim (= konsentrasi S pada saat
V=1/2 Vmaks)
Kelas/Grup Utama Enzim berdasarkan
Tipe reaksi yang dikatalisis:

1. Oksidoreduktase
2. Transferase
3. Hidrolase
4. Liase
5. Isomerase
6. Ligase
1. Oksidoreduktase:
 mengkatalisis reaksi oksidasi-reduksi
- Bekerja pada gugus tertentu dari substrat (misal: gugus
CH-OH, gugus aldehida, gugus alkana CH-CH, gugus
amina CH-NH2, gugus sulfur dll)

- Contoh Oksidoreduktase:
* etanol dehidrogenase mengkatalisis reaksi oksidasi
etanol menjadi asetaldehida dgn bantuan kofaktor NAD
Contoh Oksidoreduktase:
* glukosa oksidase memindahkan sepasang elektron dari
substrat β-D-glukosa ke oksigen  β–glukono-δ-lakton +
H2O2
(Konversi glukosa menjadi glukono lakton dapat
dilakukan secara enzimatik atau fermentasi oleh mikroba
tertentu)
2. Transferase
 Mengkatalisis reaksi pemindahan (transfer) gugus
fungsional (gugus metil, amino, asil, fosfat, glikosil dll)
Contoh :
* Aminotransferase (pemindahan gugus amino):
Asam glutamat + asam oksaloasetat  asam α-keto
glutarat + asam aspartat

* Fosfofruktokinase (pemindahan gugus fosfat):

Fruktosa 6-fosfat Fruktosa 1,6-difosfat

ATP ADP
3. Hidrolase
 Mengkatalisis pemecahan (hidrolisis) ikatan
C-O, C-N, O-P dan C-S
Contoh :
molekul polimer pati, selulosa, pektin, protein, lipida
dll  menjadi molekul lebih sederhana (penting
pada industri !) . Contoh: protease, lipase, esterase,
glikosidase dll.
- Bekerja pada ikatan peptida, ikatan glikosidik, ikatan
ester, ikatan eter, ikatan hidrogen-fosfat dll
- Spesifitas beragam, dari terbatas (contoh tripsin 
memecah ikatan peptida pada sisi karboksil lisin &
arginin) sd luas (selulase  memecah ikatan
glikosidik β-1,4 secara acak)
4. Liase
 Mengkatalisis reaksi pemindahan gugus H2O,
amonia atau CO2 dari substrat
Contoh: aldolase, dekarboksilase, dehidratase
pektinase
Histidin dekarboksilase:
(memindahkan gugus CO2 dari histidin yang
penting pada metabolisme asam amino)
Histidin  Histamin + CO2

Note : histamin dapat menyebabkan keracunan

pada produk hasil laut
5. Isomerase
 Mengkatalisis reaksi isomerasi bentuk cis-trans, keto-
enol, aldosa-ketosa dll (rasemase, epimerase,
isomerase cis-trans, dll)
- Contoh:
* Rasemase (perubahan bentuk D menjadi L)
Asam D-laktat  Asam L-laktat

* Glukosa isomerase (isomerisasi glukosa menjadi

fruktosa  pembuatan HFS)
Reaksi Katalitik Oleh Glukosa Isomerase
mengkatalisis isomerisasi :
- D-glukosa  D-fruktosa
- D-xilosa  D-xilulosa.
HOH2C
H HOH2C OH
H O
O
H
H OH
HO OH
HO H H
OH CH2OH
HO H
H

D - Glukosa D - Fruktosa

HO H CH2OH
H O
H O
H H OH

HO OH
HO H H OH
OH
HO H
H
D - Xilosa D - Xilulosa
6. Ligase
 mengkatalisis reaksi sintesis dgn penggabungan
dua mol substrat dgn bantuan ATP = Sintetase
(dengan membentuk ikatan C-O, C-S, C-N, C-C dll)
- Contoh :
Amino asil tRNA sintetase, glutamin sintetase, piruvat
karboksilase dll
Glutamat + NH3 + ATP  Glutamin + ADP + Pi
glutamin sintetase
Karakteristik Beberapa Enzim Penting

 Penting diketahui untuk aplikasinya :

- α-Amilase

- Amiloglukosidase (AMG)

- Selulase

- Protease

- Lipase
 α-Amilase (1,4-α-glukan-glukanohidrolase)

 Enzim -amilase terdistribusi secara luas di alam, terdapat pada

hewan, tumbuhan dan mikroba.

 Menghidrolisis pati secara acak melalui bagian dalam (endo-hydrolase/

endoenzim) dgn memutus ikatan -(1,4) glikosidik pada amilosa,
amilopektin dan glikogen
Enzim -amilase umumnya diisolasi dari Bacillus amyloliquefaciens,
B. licheniformis, Aspergillus oryzae dan A. niger

 pH optimum untuk enzim ini sekitar 6 dengan suhu optimum 60 oC

 Enzim -amilase yang dibuat oleh Novo Industri A/S antara lain
adalah Termamyl yang memiliki ketahanan terhadap suhu sekitar
95 – 110oC

 Dosis enzim yang digunakan sekitar 0,5 – 0,6 kg/ton pati atau
1500 U/kg substrat kering (baca brosur/label kemasan)
 Hidrolisis amilosa oleh α-amilase terjadi melalui dua
tahap :

1. Tahap pertama adalah degradasi polimer pati menjadi maltosa dan

maltotriosa yang terjadi secara acak. Degradasi ini sangat cepat
diikuti dengan menurunnya viskositas secara cepat.

2. Tahap kedua relatif lebih lambat dengan pembentukan glukosa

dan maltosa sebagai hasil akhir.

 Hidrolisis amilopektin oleh α-amilase menghasilkan

glukosa, maltosa dan berbagai jenis α-limit dekstrin, yaitu
oligosakarida yang terdiri dari empat atau lebih residu glukosa yang
mengandung ikatan α-1,6- glikosidik.

amilosa

amilopektin
β-Amilase

 merupakan eksoenzim yang melepaskan maltosa dari

ujung non pereduksi suatu polisakarida, dengan cara
menghidrolisis ikatan α 1,4 glikosidik

 Digunakan untuk membuat sirup maltosa

Dextranase

 Menghidrolisis dekstran (meyebabkan cairan kental)

pada ikatan α 1.6 glikosidik menjadi oligosakarida lebih
sederhana  digunakan pada penggilingan tebu untuk
pembuatan gula pasir
Amiloglukosidase (AMG) (glukoamilase, gamma-amilase)

 Eksoenzim yang memecah ikatan -(1,4) glikosidik

dengan melepaskan unit- unit glukosa dari ujung non-
pereduksi molekul amilosa dan amilopektin.

Enzim ini juga dapat memotong ikatan α-1,6-glikosidik

akan tetapi kerjanya relatif lambat  perlu penggabungan
dgn debrancing enzyme untuk mempercepat hidrolisis
pada ikatan α-1,6-glikosidik. Salah satunya adalah
penggunaan pullulanase yang dapat bekerja cepat
memotong ikatan α-1,6-glikosidik pada amilopektin
menjadi maltosa dan maltotriosa  AMG + pullulanase =
dextrozyme
 Amiloglukosidase diproduksi dalam jumlah besar oleh
kapang dan khamir  hanya Aspergillus dan Rhizopus
yang digunakan secara komersial

Suhu optimum untuk enzim amiloglukosidase berkisar

40-60oC dengan pH optimum sekitar 3-8

 Digunakan dengan dosis berkisar 0,65 – 0,80 liter enzim

per ton pati dengan dosis sebesar 200 U/kg pati

 Amiloglukosidase yang berasal dari Novo tersedia

dalam bentuk cair
 Selulase
-Merupakan kompleks enzim yg terdiri dari beberapa enzim
yg bekerja secara bertahap atau bersama-sama meng-
uraikan selulosa menjadi glukosa

Endo-β-1,4-
glukanase
β-1,4- glukan
β -1,4- glukanase
selobiohidrolase
Ekso-β-1,4-
Selulase glukanase β-1,4- glukan
glukohidrolase
β -1,4- glukosidase

Selulosa :
berasosiasi dgn lignin & hemiselulosa = holoselulosa/
lignoselulosa
 Homopolimer glukosa dengan ikatan β-1,4 glikosidik
β -1,4- glukanase :
 Menghidrolisis ikatan β -1,4 glikosidik secara acak terutama pada
daerah amorf serat selulosa
Tidak menyerang selobiosa, tapi menghidrolisis selodekstrin dan
selulosa yg telah dilunakkan dgn asam fosfat & selulosa yg telah
disubstitusi (e.g CMC)

β -1,4- glukosidase :
Menghidrolisis selobiosa dan selo-oligosakarida  glukosa
(tidak menyerang selulosa/selodekstrin)

β-1,4- glukan selobiohidrolase :

Menyerang ujung rantai selulosa non pereduksi  selobiosa
 Tak menyerang selulosa yg disubstitusi & selobiosa

β-1,4- glukan glukohidrolase :

 Menyerang ujung rantai selulosa non pereduksi  glukosa
 Mekanisme hidrolisis selulosa secara enzimatis

Cx Cl C
Selulosa Selulosa reaktif Selobiosa Glukosa
Cx : glukanase ; Cl : selobiohidrolase ; C : β-glukosidase

Mikroba Penghasil Selulase : a.l.

- Trichoderma reseei  aktivitas lbh tinggi
- Clostridium thermocellum

Aplikasi Produksi Bioetanol dari Lignoselulosa

Bahan Baku : Bagas (limbah tebu pabrik gula)

Perlakuan awal : + Kapang (Ceriporiopsis subvermispora, Lentinus
edodes & Pleurotus ostreatus & perebusan
Hidrolisis kombinasi 3 enzim (selulase, selobiase & xylanase)
Fermentasi serempak Saccharomyces cerevisiae
 Hasil etanol 36,4 %

Produksi Bioetanol dari bagas dgn hidrolisis selulase, α-amilase & AMG
Lactase

Lactase is a glycoside hydrolase enzyme that cuts lactose into

it's constituent sugars  galactose and glucose.

Without sufficient production of lactase enzyme in the small

intestine, humans become lactose intolerant, resulting in
discomfort (cramps, gas and diarrhea) in the digestive tract
upon ingestion of milk products.

Lactase is used commercially to prepare lactose-free

products, particularly milk, for such individuals. It is also used
in preparation of ice cream, to make a creamier and sweeter-
tasting product.

Lactase is usually prepared from Kluyveromyces sp. of yeast

and Aspergillus sp. of fungi.
Protease

 Memutus protein dg cara menghidrolisis ikatan peptida

 Aplikasi terutama pada industri deterjen dan keju (rennin)
 Penggunaan lain : industri farmasi, industri kulit,
pembuatan protein hidrolisat, industri makanan, industri film
dan penanganan limbah

 Mikroba penghasil protease :

Protease alkali :
- Stabil terhadap suhu tinggi, pH 9-11, chelating agent
- Mikroba penghasil : Bacillus licheniformis, B.
amyloliquefaciens, B.megaterium, B.pumilus -
Streptococcus griceus, S. fradiae, S.rectus - Aspergillus
niger, A. sojae, A.oryzae, A. flavus
 Protease Netral :
- Relatif tidak stabil thd pH, suhu & dpt diinaktivasi oleh
protease alkali
- Aplikasi : industri kulit & industri makanan (crackers, roti
dll)

Protease Asam
- Aktif pada suasana asam, contoh : rennin & pepsin
(pH opt 2-3)
- Aplikasi :- Protease dari Aspergillus  untuk memecah
gluten pada pemanggangan

Rennin
- Menggumpalkan kasein susu pada pembuatan keju
- Mikroba penghasil : S. lactis, S. cremoris, S. thermophilus,
Lactobacillus bulgaricus, L. helveticus
Lipase (Gliserol ester hydrolase)

 Memutus lemak menjadi di atau monogliserida dan asam

lemak

 Mikroba penghasil :
Kapang : Aspergillus, Mucor, Rhizopus,
Khamir : Candida

 pH optimum netral atau agak asam, suhu optimum

30 – 45 0C

 Aplikasi :
- farmasi : enzim pencernaan (suplemen lipase pankreas)
- Industri susu (pematangan keju)
- dll
APLIKASI ENZIM
Important Applications of α-Amylase
 Suspensi pati Produksi Sirup Glukosa &
Gelatinasi Sirup Fruktosa
Pati α-Amilase bakteri
(95-105 0C) (pH 6.0-6.6, 105oC, 5 mnt atau 95oC 1-2 jam

Sakarifikasi
Maltodekstrin
Sirup Glukosa
α-Amilase kapang
Sakarifikasi + glukoamilase
pH 5.0, 55oC, 42
Glukoamilase atau jam
Glukoamilase+ Sakarifikasi
Pululanase Amilase kapang atau
pH 4.0-4.5, 60oC, β-amilase
48-72 jam pH 5.5, 55oC, 42 jam
Sirup Glukosa

Kolom Penukar Ion

Sirup Maltosa
Isomerisasi Glukosa isomerase
pH 7.5-8.5, 60-65oC, 2-4 jam
Sirup Fruktosa
Sirup Fruktosa Berkadar tinggi
Pemurnian
Manfaat Enzim dalam Industri
Bentuk jual beberapa enzim industri
 * Aplikasi Oksidoreduktase
Enzim Sumber Konversi yg dikatalisis Manfaat
Katalase A. niger, 2 H2O2  2 H2O + O2 -Membuang residu H2O2 (racun)
Micrococcus setelah sterilisasi dingin susu
lysodeikticus dan putih telur.
dan -Membuang peroksida dari kerja
bovine liver glukosa oksidase
- Membuang O2 dari sari buah
(kerjasama dg glukosa oksidase)
Glukosa A. niger Glukosa + O2  -Menghilangkan gula pada
Oksidase Glukono-δ-lakton + H2O2 putih telur untuk mencegah
browning dan off-flavor
selama pengeringan
Lipoksigenase Tepung kedelai Linoleat + O2  LOOH Menghilangkan bau langu kacang
Contoh Aplikasi Oksidoreduktase :

Glukosa oksidase memindahkan sepasang elektron dari substrat

β-D-glukosa ke oksigen  β–glukono-δ-lakton + H2O2

The glucose oxidase enzyme is commonly used in biosensors to

detect levels of glucose by keeping track of the number of
electrons passed through the enzyme by connecting it to an
electrode and measuring the resulting charge.
This has a possible application in the world of nanotechnology
when used in conjunction with tiny electrodes as glucose sensors
for diabetics.

http://www.indogulfgroup.com/AE%20-%20Enzymes%20-%20Glucose%20Oxidase.asp
 Glucono delta-lactone (GDL):

is a naturally-occurring food additive with the E number E575

used as a sequestrant, an acidifier, or a curing, pickling, or
leavening agent in bakery products.

- GdL ia a slow leavening agents, based on the progressive

hydrolysis of GdL to gluconic acid when water is added to the
dough mix. The gluconic acid then reacts with sodium
bicarbonate for a continuous release of carbon dioxide.)
- Also functioning as a substitute for enzymes in cheese
processing or tofu manufacturing . GdL is acoagulant in silken
tofu, (tofu with a smooth texture similar to yoghurt and a high
water content). GdL is also combined with calcium sulphate or
calcium chloride to produce a silken tofu with a firmer texture.

http://www.jungbunzlauer.com/media/uploads/pdf/Gluconates/Glucono-delta-Lactone_2008.pdf
 Glucono-delta-lactone (GDL) Utilisation

GDL is used for various technological purposes with regard to

foodstuffs:
• for the regulation of acidity and colour in food
• for the support of antioxidants and preservatives
(suppress the growth of unwanted or pathogenic microrganisms)
• for sausage and canned fish (as an accelerator of maturation)
• for shellfish and processed vegetables (as a stabiliser of colour)
• in baking powder (to facilitate the consistent production of acidity
and so to provide constant support of baking procedures)

http://www.gmo-compass.org/eng/database/e-numbers/294.glucono_delta_lactone.html
Glucono delta-lactone (GDL)
Curing Accelerator in Meat Products

• In curing: red colour formation process of sausages

are highly dependent on the pH .
• When using GdL in scalded sausages, the curing
process is both accelerated and made more
complete, resulting in a brighter red colour of the
meat product.
 GdL thus allows to reduce both the addition of
nitrite by up to 30% and the residual level of nitrite
by up to 75%, making the product safer.

• GdL enhance the antimicrobial action of lactates in

pâtés, to lower the pH of poultry products to
remedy their pinking.
Chelating Agent in Seafood

• GdL, as an outstanding copper and iron chelating agent, helps

inhibiting melanosis by depriving the enzymes causing the
browning of the metal ions needed for their activation.
• The combination with sulphites in the pre-freezing dip of
frozen shrimps or in the blanching brine of canned shrimps
allows to come away from calcium disodium EDTA and to
reduce the sulphite use by 50-90%.
 Karbohidrase
Enzim Sumber Konversi yang dikatalisis Manfaat
α-amilase Barley malt, pati, glikogen + H2O  Hidrolisat pati pada industri
fungi, bakteri Dekstrin (likuifikasi) minuman, pembuatan roti,
dextrinized starch, makanan bayi
β-amilase Wheat, barley, pati, glikogen + H2O  Industri roti dan minuman ber-
malt, bakteri Maltosa β-limit dextrin alkohol: sumber C utk produksi
(bacillus) (ikatan α-1,4) CO2 dan alkohol, sirup maltosa
β-glukanase A. niger, pati-D-glukan + H2O  Degumming pada
B. subtilis, oligosakarida, glukosa industri minuman
barley malt (ikatan β-1,3 dan β-1,4) beralkohol
Glukoamilase A. niger, pati, glikogen + H2O  Sakarifikasi pati
(amilogluko- A. oryzae, glukosa (ikatan α-1,4) terlikuifikasi
sidase, AMG) R. oryzae
Debranching A. aerogenes, amilopektin + H2O  amilosa rantai Oligosakarida (amilosa
enzymes Pseudomonas pendek menghidrolisis ikatan α-1,6 rantai pendek)
(pullulanase,
isoamylase)
Inulinase Kluyvero-myces Polifruktan (inulin) + H2O  fruktosa HFS
Isomerase Glukosa < --- > fruktosa HFS
Xilosa < --- > xilulosa
Selulase A. niger, T. viride Selulosa + H2O  β-dekstrin (ikatan Klarifikasi jus buah,
β-1,4) meningkatkan ekstraksi
minyak atsiri, produksi
glukosa dari bahan
selulosa
Hemiselulase A. niger Hemiselulase + H2O  β-dekstrin Memudahkan pengelupas-
(ikatan β-1,4) dari locust bean gum, an kulit kopi, meningkat-
guar gum kan germinasi biji-bijian dll
Invertase (sukrosa Saccharomyces Sukrosa + H2O  glukosa Mengkatalisis pembentukan gula
hidrolase) + fruktosa (gula invert) invert, mencegah kristalisasi
produk permen
Laktase A. niger, Laktosa + H2O  Hidrolisis laktosa dalam produk
(Β- galaktosidase) A. oryzae galaktosa + glukosa susu, meningkatkan kemanisan
produk, mencegah kristalisasi
laktosa, low-lactose milk untuk
penderita lactose intolerance
Pektinase, terdiri A. niger, A. *Poligalakturona-se Ditambahkan pada klarifikasi
poligalakturonase, oryzae menghidrolisis β-D-1,4 dan filtrasi jus dan wine,
pektin metil galakturonida mencegah penggumpalan pada
esterase, pektat liase *Pektin metilesterase jus buah konsentrat, mengatur
menghilangkan gugus cloud rentention pada jus, dll.
metil pada pektin
• Apple juice is one of the fruit juice which is manufactured by
maceration and pressing of apples.
• The expelled juice is further processed by centrifugal and
enzymatic clarification to remove the pectin and starch.

http://www.tastydays.com/drinks/juice/apple-juice
 Pulp and Paper Industry

Type Enzyme Function

Cellulase Breaks down cellulose, a fibre found in the cell walls of all
plants and trees. Cellulose is the basic raw material used
to make products such as paper, cotton, and other textiles
Hemicellulase Breaks down hemicellulose, another plant sugar that is
not as complex as cellulose and is easier to break down
Xylanase Breaks down xylan, a gummy sugar present in the cell
walls of plants and trees. This enzyme type is used
primarily in the wood and pulp industry

What are Enzymes? http://www.biobasics.gc.ca/english/View.asp?x=792

 Enzyme Biotechnology and Biofuels

• Enzymes may be used to help produce fuels from renewable sources

of biomass. Such enzymes include cellulases, which convert cellulose
fibres from feedstocks like corn into sugars. These sugars are
subsequently fermented into ethanol by microorganisms.

• A new process called “Simultaneous Saccharification and Fermentation

has greatly improved ethanol production efficiency. In this new process,
cellulase enzymes and fermentation microorganisms are combined in a
single reaction mixture to produce ethanol in one step .
 * Aplikasi Protein Hidrolase
Enzim Sumber Konversi yang dikatalisis Manfaat
Bromelin Nanas Hidrolisis protein dan polipeptida -Mengempukkan daging,
menjadi peptida dengan BM -Meningkatkan ekstraksi minyak
rendah dan protein dari tanaman dan
Ficin Ficus sp. Protease dari tanaman. hewan,
Hidrolisis polipeptida, amida dan -Pembuatan hidrolisat protein
Papain Pepaya -Pengolahan protein ikan
ester (leu, gly) menjadi peptida
dengan BM rendah
Protease Aspergillus Hidrolisis protein dan polipeptida -Meningkatkan warna
Bacillus menjadi peptida dengan BM dan flavor roti,
rendah -Mengempukkan daging
-Meningkatkan dispersitas susu
bubuk & meningkatkan stabilitas
susu evaporasi
Pepsin Porcine, Hidrolisis polipeptida (asam Bovine pepsin  rennet
Perut amino aromatik, dikarboksilat) extender dan koagulan
hewan menjadi peptida dengan BM susu (keju), produksi
rendah hidrolisat protein
Tripsin Pankreas Hidrolisis polipeptida (ggs Menekan oksidasi flavor di
hewan karboksil L-arg, L-lys) menjadi susu; produksi hidrolisat
peptida dengan BM rendah protein
Rennet Perut hewan Selektivitas tinggi terhadap Koagulasi susu dalam
ruminansia; kasein susu, mengkoagulasi pembuatan keju,
Mucor miehei, M. susu meningkatkan flavor dan
Pusilus, Endothia tekstur pada pematangan
parasitica keju
Enzymes as Detergent

• Enzymes have also been used in the detergent industry since

the mid 1960’s and are probably the best-known application of
industrial enzymes especially in laundry products- the so-called
“biological” washing powders, liquids and tablets.

• The main enzyme used : protease - group of hydrolases acting

on peptide bonds, belongs to EC 3.4 group.
• Used for removing of protein contamination  acts on organic
stains such as grass, blood, egg and human sweat.

http ://enzymes.me.uk
However, it has become more common in recent years to
include a mixed of enzyme activities including lipases and
amylases.

 Lipases (group of esterases (EC 3.1) disintegrated lipids.

Allow to remove fatty stains  effective on stains resulting
from fatty acids such as oils and fats (and lipsticks!)
 Amylase help remove starchy food deposits.

 Cellulase : more recently, cellulase used as a colour

enhancing and ‘anti-bobbling’ washing powders .
 Cellulases is removed detached cellulose fibrils, which cause
a progressive dulling of the colour as dirt is trapped on the
rough surface of the fabric.
Formulasi Enzim Deterjen

• Nilai pH deterjen 10  protease alkalin dari Bacillus sp

• Bentuk awal : dry powder  pekerja alergi & gangguan kulit

• Formulasi :
enkapsulasi agar debu tak beterbangan, partikel larut
cepat, tak menghasilkan bau & warna, daya simpan lebih
baik, melindungi thd komponen lain e.g bleaching agent
*Proses Enkapsulasi Protease metode NOVO

• bentuk : granula menggunakan “pelletising drum”

• untuk mencegah pelekatan pada dinding mixer & menjaga ukuran
granula, ditambahkan serat selulosa
• merk : Granule-T

*Proses Enkapsulasi Protease metode Gist-Brocades

• Cairan fermentasi (broth)  filtrasi  presipitasi enzim  filtrasi 

pengeringan  disaring  powder

• Formulasi :
enzim powder dicampur aditif etoxylated-C18-fatty alcohol
 “prill tower”  bentuk bola dilapis agar tidak berdebu selama
penanganan (pelapis (minyak parafin atau PEG)
 merk : Maxatase dan Maxacal (pH opt ≥10)
Komposisi Deterjen Enzim
 Industri Keju

Rennet
• Milk contains proteins, specifically caseins, that maintain its
liquid form.
• Proteases are enzymes that are added to milk during cheese
production, to hydrolyze caseins, specifically kappa casein, which
stabilizes micelle formation preventing coagulation.
• The most common enzyme isolated from rennet is chymosin.,
which can be obtained from several other animal, microbial or
vegetable sources, but indigenous microbial chymosin (from
fungi or bacteria) is ineffective for making cheddar and other
hard cheeses.

http://enzymes.me.uk/enzyme/enzymes-in-industry
Produksi Keju
Lipases
• Lipases are used to break down milk fats and give characteristic
flavours to cheeses.
• Stronger flavoured cheeses, for example, the italian cheese,
Romano, are prepared using lipases.
• The flavour comes from the free fatty acids produced when milk
fats are hydrolyzed.
• Animal lipases are obtained from kid, calf and lamb, while
microbial lipase is derived by fermentation with the fungal species
Mucor meihei.
• Although microbial lipases are available for cheese-making, they
are less specific in what fats they hydrolyze, while the animal
enzymes are more partial to short and medium-length fats.
• Hydrolysis of the shorter fats is preferred because it results in the
desirable taste of many cheeses. Hydrolysis of the longer chain
fatty acids can result in either soapiness, or no flavour at all.
• Milk contains a number of different types of proteins, in addition to
the caseins. Cow milk also contains whey proteins such as lactalbumin
and lactoglobulin.
• The denaturing of these whey proteins, using proteases, results in a
creamier yogurt product. Destruction of whey proteins is also essential
for cheese production.

• During production of soft cheeses, whey is separated from the milk

after curdling, and may be sold as a nutrient supplement for body
building, weight loss, and lowing blood pressure, among other things.
• There have even been reports of dietary whey for cancer therapies,
and having a role in the induction of insulin production for those with
Type 2 diabetes.
• Proteases are used to produce hydrolyzed whey protein, which is whey
protein broken down into shorter polypeptide sequences. Hydrolyzed
whey is less likely to cause allergic reactions and is used to prepare
supplements for infant formulas and medical uses.
 * Lipase

Enzim Sumber Konversi yang dikatalisis Manfaat

Lipase Jaringan Hidrolisis triasil gliserol, Lipase hewan: meningkatkan
pankreas ester asam lemak, flavor pada industri keju,
hewan, menghasilkan mono- dan di- lypolized butter oil
A. Niger, asilgliserol, gliserol dan asam Lipase mikrobial: menghidrolisis
A. oryzae lemak bebas lipid (konsentrat minyak ikan)

This oil contains good Omega 3 polyunsaturated fats, particularly DHA, the
most important of the Omega 3 fats, and EPA, also important.
Cocoa Butterv Substitute
• Cocoa butter is extracted from the beans of the Theobroma
cacao tree and used to make chocolate and other foods,
pharmaceuticals and cosmetic ointments.

• The expense of cocoa butter has led to the production of

cocoa butter substitutes.
• The substitutes (known commercially as CBS) generally are
made from plant oils, such as palm kernel oil, sunflower oil,
cottonseed oil, soybean oil and coconut oil.
Cocoa Butter Substitute

• Lipase-catalysed trans-esterification reactions that are 1,3-

specific are utilized for making cocoa butter substitutes, using
cheaper mid-fraction of palm oil and stearic acid.

 The mid-fraction of palm oil contains 1,3-dipalmitoyl-2-

monoolein (POP) as the major triglyceride
 The main constituents of cocoa butter 1-(3)-palmitoyl-3-(1)-
stearoyl-2-monoolein (POS) and 1-(3)-distearyl-2-monoolein
(TOS)
 Therefore, by transesterification catalyzed by lipozyme,
between POP and stearic acid or POP and tristearin,
valuable equivalents of cocoa butter have been made

http://www.ias.ac.in/currsci/jul10/articles18.htm
Lipases for Pharmaceutical Application

Microbial lipases are used to enrich PUFAs from

animal and plant lipids, and their mono and
diacylglycerides are used to produce a variety of
pharmaceuticals. PUFAs are increasingly used as
food additives, pharmaceuticals and nutraceuticals

Microbial lipases are used to obtain PUFAs from animal and plant
lipids, such as tuna oil and borage oil. Free PUFAs and their mono
and diacylglycerides are subsequently used to produce a variety of
pharmaceuticals

Liposomes are used in the medical field to optimize the action of

drugs by transporting them to target areas, thus circumventing drug
waste inactivation and anatomical barriers

http://nopr.niscair.res.in/bitstream/123456789/3016/1/IJBT%206%282%29%20141-158.pdf
Enzyme treated Lecithin

• Lecithin can easily be extracted chemically (using hexane)

or mechanically from soy bean etc.
• It used as an emulsifier and to prevent sticking, for
example in non-stick cooking spray.

• To modify the performance of lecithin to make it suitable

for the product to which it is added, it may be hydrolysed
enzymatically. In hydrolysed lecithins, a portion of the
phospholipids have one fatty acid removed by
phospholipase.  improved through strengthening the
hydrophobicity
Enzim dalam Teknologi Rekayasa Genetika

 Pembuatan DNA Rekombinan

- Memotong gen yang akan ditranlasikan,
- Membuat ujung fragmen DNA yg sesuai, sehingga dapat
digunakan dg fragmen lain yang dikehendaki
- Penyambungan fragmen-fragmen DNA membentuk DNA Rekombinan

1. Endonuklease Restriksi
 memotong DNA
2. DNA Ligase
 untuk proses replikasi & menyambung fragmen DNA dg
fragmen lain atau kloning membentuk DNA Rekombinan
3. Nuklease
 menguraikan komponen nukleotida dari DNA (berperan
pd proses replikasi)
4. RNA Polimerase
 berperan pada proses transkripsi DNA  mRNA
Enzymes in molecular biology

This is never ending story. Many of enzymes used in molecular

biology directly in bacterial cells to synthesise necessary
compounds.

Bacterial and yeasts cells can be used for any protein and
peptide biosynthesis. Some of these bacterial enzymes can be
used in vitro to perform the same biological processes, from
which are most important enzymes for DNA/RNA manipulation.

http ://enzymes.me.uk
Therapeutic Enzymes
Daftar Pustaka
 2011, Mata Kuliah Satuan Proses, Departemen Teknologi Industri
Pertanian, Fakultas Teknologi Pertanian, Institut Pertanian Bogor,
k002.kiwi6.com/hotlink/9v58ohmgjx/kuliah_xi_enzim_final.ppt, diakses 12
Juni 2012