Industrial Applications

of Enzymes

z Carbohydrates
„ Essentialcomponents of all living
organisms and most abundant.
„ Chemical & Energy Balance in the
Biosphere
Photosynthesis: Conversion of
atmospheric CO2 into carbohydrate
nCO 2 + nH 2 O ⇔ nO 2 + ( CH 2 O) n

Respiration: Carbohydrates are

ingested by higher animals to yield
energy and CO2 is released
 Monosaccharides
(CH2O)n

z Derivatives of
„ Polyhydroxy (-OH) aldehyde (-CHO)
– Aldose
„ polyhydroxy (-OH) ketone (=CO)
–Ketose
zn =3~8
„ triose
„ tetrose
„ pentose
„ hexos
„ heptose
 Asymmetric carbon (AC):
Carbon with 4 different substituents

z Number of stereo isomer = 2AC

Aldohexos has 4 AC.

It has 16 possible isometric forms.
„ 8 L form: OH on the left side of
the highest AC
„8 D form: OH on the right side
Glucose

Glucopyranose
Haworth Projection

α-D-glucopyranose
 Fructose

CH2OH
C=O O
HOCH HOH2C OH
HCOH H CH2OH
HCOH
CH2OH

D-fructose b-D-fructose
(b-D-fructofuranose)
 Disaccharides

CH2OH CH2OH CH2OH

O O O O
HOH2C OH OH
O
O CH2OH

Sucrose Lactose (b form)

CH2OH CH2OH CH2OH CH2OH

O O O O
OH
O
O OH

Maltose (a form) Cellobiose (b form)

z Amylose

z Amylopectin

z Cellulose
 Starch Hydrolysis
z Corn (or Glucose) Syrup
„ Ethanol Fermentation
z High Fructose Corn Syrup
– 42% fructose (Similar sweetness as
sugar)
„ Fructose
– 2.3 times sweeter than glucose
– 1.4 times sweeter than sugar
„ Usage
Soft drinks, Canned fruits, Juice, Ice
cream, etc
„ Source:
Corn
– Low costs
– Useful by-products: oil, gluten, corn
meal, dry starch, fiber, etc.
 Corn Refining Process
α−amylase

Liquefaction
tank Flash Cooler
Starch 80 - 150 °C
Slurry pH 6 - 7 Filtration
3 hrs.
15% D.E. Carbon Refining
Saccharifi-
cation tank Ion Exchanger
Fungal
Glucoamylase 55 - 60 °C
pH 4 - 4.5 95% Evaporator
40 to 80 hrs. D.E.
Glucose Syrup

Isomerization
with Immobilized
Glucose Isomerase
Cooler
60 - 65 °C
pH 7 - 8.5
30 mins. Filtration

Carbon Refining

Ion Exchanger
HFCS
Evaporator
42% Fructose
 Cellulose Hydrolysis

z Cellulose:
Most abundant in nature
z Typical Dry Wood
„ Cellulose 41%
„ Hemicellulose 26%
– Lower MW than cellulose
– Contains pentoses
„ Lignin 28%
– Cementing stiffening polymer
– Polymers of phenolic compounds
„ Other 5%
 Structure of Cellulose Fibers

Cellulose Moleculres

Elementary
fibrils

Microfibrils

Elementary
fibril
Cell wall
component Polyoses
of wood
Lignin
 Economic Bottlenects to
Biomass Utilization
z Availability in large quantity
z High costs of pretreatment
z Total biomass utilization
z Low solid reaction
z High cost of separation

Economic Bottlenects for

Enzyme Hydrolysis
z Slow reaction rate
z High costs of enzymes
z High costs of pretreatment
Attrition Bioreactor (ABR)
 Hydrolysis of Newsprint in ABR

700

600
Reducing Sugar (mg/g DM)

500

400

300

200
A B R w ith 0 .4 7 6 c m S .S . b a lls
S T R w ith 1 w k b a ll m illin g
100
S T R

0
0 1 2 3 4 5 6
T im e ( h o u r s )
 Enzyme Stability in ABR

1 .2
Relative Enzyme Activity

0 .8

0 .6

0 .4
S e a le d R e a c to r
0 .2
O p e n R e a c to r
0
0 4 8 12 16 20
T im e (h o u rs )