Aktivitas Enzim

TIN 321 / 2 (2-0)
m.k. SATUAN PROSES
XI. ENZIM
Proses Konversi Pada Bahan Yang
Memanfaatkan Enzim Sebagai
Katalis
DEPARTEMEN TEKNOLOGI INDUSTRI PERTANIAN
FAKULTAS TEKNOLOGI PERTANIAN
INSTITUT PERTANIAN BOGOR
2011
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 ” = bag 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  U/mL
• Aktivitas Spesifik  U/mg protein enzim

ukuran kemurnian
pH optimum
Laju reaksi
Laju. reaksi
Laju maksimum
Laju maksimum
suhu optimum
pH
suhu
Vm
2
Laju maksimum
Pembentu
kan
Produk
Vm
b. Pengaruh suhu
Km
[substrat]
Vm: Laju Reaksi maksimum
Km: konstanta Michaelis
c. Pengaruh [substrat]
Enzim1 aktivator
enzim
Waktu
d. Pengaruh aktivator
Pembentu
kan
Produk
Laju. reaksi
a. Pengaruh pH
enzim
Enzim1 inhibitor
Waktu
d. Pengaruh inhibitor
Pengaruh Substrat
E+S
K1
K--1
ES
Vmaks S 
 V 
K m  S 
K2
E+P
 Persamaan Michaelis-Menten
Saat V = ½ Vmaks  Km = [S]
Vmaks
½ Vmaks
Km
→[S]
Line Weaver-Burk  1  K m . 1 
V
Vmaks
a
1
V
1
S 
1
Vmaks
b
Km
Slope 
Vmaks
Vmaks

1
Km
1
S 
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 (e.g 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, ketoenol, 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
H
H
HO
O
H
OH
OH
H
HO
OH
HOH2C
O
H
OH
CH2OH
H
HO
H
D - Glukosa
HO
H
D - Fruktosa
CH2OH
H
H
O
O
H
H
HO
H
HO
OH
OH
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 60oC
 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 nonpereduksi 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 menguraikan selulosa menjadi glukosa
Endo-β-1,4glukanase
β -1,4- glukanase
Ekso-β-1,4glukanase
Selulase
β-1,4- glukan
selobiohidrolase
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
Selulosa
Cx
Selulosa reaktif
Cl
Selobiosa
C
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
Produksi Sirup Glukosa &
Sirup Fruktosa
Suspensi pati
Gelatinasi
Pati
(95-105 0C)
α-Amilase bakteri
(pH 6.0-6.6, 105oC, 5 mnt atau 95oC 1-2 jam
Maltodekstrin
Sakarifikasi
Glukoamilase atau
Sakarifikasi
Glukoamilase+
Pululanase
pH 4.0-4.5, 60oC,
48-72 jam
Sirup Glukosa
Sakarifikasi
α-Amilase kapang
+ glukoamilase
pH 5.0, 55oC, 42
jam
Sirup Glukosa
Amilase kapang atau
β-amilase
pH 5.5, 55oC, 42 jam
Kolom Penukar Ion
Isomerisasi
Sirup Maltosa
Glukosa isomerase
pH 7.5-8.5, 60-65oC, 2-4 jam
Sirup Fruktosa
Pemurnian
Sirup Fruktosa
Berkadar tinggi
Bentuk jual beberapa enzim industri
* Aplikasi Oksidoreduktase
Enzim
Sumber
Konversi yg
dikatalisis
Katalase
A. niger,
2 H 2 O2 
Micrococcus
2 H 2 O + O2
Lysodeikticus
&
bovine liver
Manfaat
-Membuang residu
H2O2 (racun) setlh
sterilisasi dingin susu
dan putih telur,
-Membuang peroksida
dari kerja glukosa
oksidase
- Membuang O2 dari sari
buah (kerjasama dg
glukosa oksidase)
Glukosa A. niger
Oksidase
Glukosa + O2 
Glukono-δ-lakton +
H2O2
-Menghilangkan gula
pada putih telur untuk
mencegah browning
dan off-flavor selama
pengeringan
Lipoksige Tepung
-nase 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,
fungi, bakteri
pati, glikogen + H2O 
Dekstrin
(likuifikasi)
Hidrolisat pati pada
industri minuman,
pembuatan roti,
dextrinized starch,
makanan bayi
β-amilase
Wheat,
barley
malt, bakteri
(bacillus)
pati, glikogen + H2O 
Maltosa β-limit dextrin
(ikatan α-1,4)
Industri roti dan
Minuman
beralkohol :
sumber karbon
untuk produksi CO2
dan alkohol, sirup
maltosa
β-glukanase
A. niger,
B. subtilis,
barley malt
pati-D-glukan + H2O 
oligosakarida, glukosa
(ikatan β-1,3 dan β-1,4)
Degumming pada
industri minuman
beralkohol
Glukoamilase
(amiloglukosidase,AMG)
A. Niger,
A. oryzae,
R. oryzae
pati, glikogen + H2O 
glukosa (ikatan α-1,4)
Sakarifikasi pati
terlikuifikasi
Debranching
enzymes
(pullulanase,
isoamylase)
A. aerogenes,
Pseudomonas
amilopektin + H2O 
amilosa rantai
pendek
menghidrolisis
ikatan α-1,6
Oligosakarida
(amilosa rantai
pendek)
Inulinase
Kluyveromyces
Polifruktan (inulin) +
H2O  fruktosa
HFS
Glukosa < --- >
fruktosa
Xilosa < --- > xilulosa
HFS
Isomerase
Selulase
A. niger,
T. viride
Selulosa + H2O  βdekstrin (ikatan β1,4)
Klarifikasi jus buah,
meningkatkan
ekstraksi minyak
atsiri, produksi
glukosa dari bahan
selulosa
Hemiselulase
A. niger
Hemiselulase + H2O
 β-dekstrin (ikatan
β-1,4) dari locust
bean gum, guar gum
Memudahkan
pengelupasan kulit
kopi, meningkatkan
germinasi biji-bijian
dll
Sukrosa + H2O  Mengkatalisis
glukosa + fruktosa pembentukan gula
(gula invert)
invert, mencegah
kristalisasi produk
permen
Invertase
(sukrosa
hidrolase)
Saccharomyces
Laktase
(Βgalaktosidase)
A. niger, A. Laktosa + H2O 
oryzae
galaktosa +
glukosa
Pektinase, terdiri A. niger, A.
dari
oryzae
poligalakturonase, pektin metil
esterase, pektat
liase
*Poligalakturonase menghidrolisis
β-D-1,4
galakturonida
*Pektin
metilesterase
menghilangkan
gugus metil pada
pektin
Hidrolisis laktosa dalam
produk susu,
meningkatkan
kemanisan produk,
mencegah kristalisasi
laktosa, low-lactose
milk untuk penderita
lactose intolerance
Ditambahkan pada
klarifikasi dan filtrasi
jus dan wine, mencegah
penggumpalan pada jus
buah konsentrat,
mengatur cloud
rentention pada jus, dll.
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
Bromelin
Nanas
Hidrolisis protein dan
polipeptida menjadi
peptida dengan BM
rendah
Ficin
Ficus sp.
Papain
Pepaya
Protease
Aspergillus
Bacillus
Manfaat
-Mengempukkan daging,
-Meningkatkan ekstraksi
minyak dan protein dari
tanaman dan hewan,
Protease dari tanaman. -Pembuatan hidrolisat
protein
Hidrolisis polipeptida,
-Pengolahan protein ikan
amida dan ester (leu,
gly) menjadi peptida
dengan BM rendah
Hidrolisis protein dan
polipeptida menjadi
peptida dengan BM
rendah
-Meningkatkan warna
dan flavor roti,
-Mengempukkan
daging
-Meningkatkan
dispersitas susu bubuk &
meningkatkan stabilitas
susu evaporasi
Pepsin Porcine,
Perut
hewan
Hidrolisis polipeptida
(asam amino
aromatik,
dikarboksilat) menjadi
peptida dengan BM
rendah
Bovine pepsin 
rennet extender
dan koagulan susu
(keju), produksi
hidrolisat protein
Tripsin Pankreas
hewan
Hidrolisis polipeptida
(ggs karboksil L-arg,
L-lys) menjadi peptida
dengan BM rendah
Menekan oksidasi
flavor di susu;
produksi hidrolisat
protein
Rennet Perut hewan
ruminansia;
Mucor miehei,
M. Pusilus,
Endothia
parasitica
Selektivitas tinggi
Koagulasi susu
terhadap kasein susu, dalam pembuatan
mengkoagulasi susu
keju, meningkatkan
flavor dan tekstur
pada pematangan
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
pankreas
hewan,
A. Niger,
A. oryzae
Hidrolisis triasil
gliserol, ester asam
lemak, menghasilkan
mono- dan diasilgliserol,
gliserol dan asam
lemak bebas
Lipase hewan :
meningkatkan
flavor pada industri
keju, lypolized
butter oil
Lipase mikrobial :
menghidrolisis
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-2monoolein (POP) as the major triglyceride
 The main constituents of cocoa butter 1-(3)-palmitoyl3-(1)-stearoyl-2-monoolein (POS) and 1-(3)-distearyl-2monoolein (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