Starch

Application Sheet

Efficient liquefaction of
starch
Alpha-amylases are widely used by starch processors to unlock
the dextrose components in a liquefaction process.
Whether the end product is high fructose syrup, dextrose,
maltose, maltodextrin, or polyols, the process starts with an
alpha-amylase that can be applied under a wide range of
processing conditions.

Novozymes' alpha-amylase range provides the right tools for

starch liquefaction.

Benefits
Novozymes offers a range of alpha-amylases that enables the starch processor
to optimize the liquefaction process in terms of cost, quality, and efficiency.
The key features offered by the liquefaction product range are:

• Efficient DE development – when high DE development in a short time is a

key parameter
• High dextrose yield – with minimal by-product formation
• Fast viscosity reduction – enabling high dry substance levels
• Low color formation – reducing refinery costs
• Savings on chemicals and ion exchange costs by eliminating calcium addition

Products
Novozymes' starch liquefaction product range consists of six different alpha-
amylases: Liquozyme® X; Liquozyme Supra; Liquozyme Supra 2.2X; Liquozyme
Supra 2.8 X; Termamyl® 120 L, Type L; and Termamyl 2X.

The most important characteristics of the products in the alpha-amylase range

are given in Table 1.
 Product
Characteristics Liquozyme® X Liquozyme Supra Termamyl® 120 L,
Liquozyme Supra 2.2X Type L
Liquozyme Supra 2.8X Termamyl 2X
DE development power under 12.5 11.1 8.2
standard conditions (see Fig. 1)
pH range 5.2–5.6 5.2–5.6 6.0–6.4
Calcium requirement (ppm) 5–15 5–15 40–70
Low maltulose formation ++++ ++++ +
Low panose With inactivation +++++ +++++ +++++
formation Without +++++ +++ +
inactivation
High DX With inactivation +++++ +++++ +++++
Without +++++ +++ +
inactivation
Sacch. With inactivation +++++ +++++ +++++
savings Without +++++ +++ +
inactivation
Low color formation ++++ ++++ +
Chemical savings +++++ +++ +
Ion exchange savings +++++ +++ +
Viscosity reduction ++++ +++++ +++++
Ease of inactivation ++ + ++++
Table 1. Characteristics of Novozymes' alpha-amylase product range.
Note 1: In addition to the above liquefaction product range, BAN® 480 L is also available for starch
liquefaction.
Note 2: DE development power is measured as DE/120 minutes under standard liquefaction conditions
(see Fig. 1).
Note 3: Best choice = +++++.

Termamyl 2X gives 2 times the performance of Termamyl 120 L, Type L. The

dose can therefore be reduced. Likewise, Liquozyme Supra 2.2X gives 2.2 times
the performance of Liquozyme Supra. Liquozyme Supra 2.8X gives 2.8 times
the performance of Liquozyme Supra.

For more information on the above-mentioned products, visit our Customer

Center at www.novozymes.com. Other product variants may be available on
request.

Performance
DE development
DE development in liquefaction is determined by enzyme dosage, process
conditions, and enzyme efficiency. The difference in efficiency between
Termamyl 120 L, Type L, Liquozyme X, and Liquozyme Supra is shown in Figure
1.
 Termamyl® 120 L, type L Liquozyme® X Liquozyme Supra

14

12
DE (Fehling), %
10

0
0 30 60 90 120

Min.

Fig. 1. DE development. Liquefaction with Termamyl® 120 L, Type L (0.4 kg/t DS; 70 ppm Ca; pH
6.2), Liquozyme X (0.4 kg/t DS; 5 ppm Ca; pH 5.5), and Liquozyme Supra (0.4 kg/t DS; 5 ppm Ca;
pH 5.5).

High dextrose yield

Maltulose (disaccharide) and panose (trisaccharide) are both undesired
conversion products that reduce the yield of dextrose in saccharification.
The content of maltulose and panose becomes apparent after saccharification
of the starch hydrolysate. The maltulose content of the final saccharified syrup
can be minimized by running the liquefaction process at a low pH. When
Liquozyme X or one of the Liquozyme Supra products are applied, the
maltulose content is typically 0–0.2% on DS.

Panose formation can be minimized by applying either Liquozyme X or one of

the Liquozyme Supra products in the liquefaction. Liquozyme X in particular
(but also to a lesser extent the Liquozyme Supra products) hydrolyzes the starch
dextrins in such a way that only small amounts of panose precursors are
produced.

With Termamyl 120 L, Type L and Termamyl 2X it is necessary to inactivate the

enzyme prior to saccharification to achieve the same low panose formation.

Reduction of maltulose and panose concentrations allows for higher glucose

content (DX) in saccharification, which is often of great value. Alternatively, a
saving in saccharification enzyme dosage could be achieved.

Viscosity reduction
When starch is gelatinized, the viscosity of the starch slurry initially increases.
However, the alpha-amylase immediately starts to hydrolyze the starch, and the
viscosity is reduced.

The viscosity profiles for three different alpha-amylases are shown in Figure 2.
 Liquozyme® Supra Liquozyme® X Termamyl® 120 L

2500
2250
2000
1750
Viscosity

1500
1250
1000
750
500
250
0
-250 0 2 4 6 8 10 12 14 16

Min.

Fig. 2. Viscosity reduction in liquefaction. Viscosity development in a 12% DS starch slurry heated from
50 °C to 95 °C (122 °F to 203 °F) with added alpha-amylase. Termamyl® 120 L, Type L (0.54 kg/t DS; 70
ppm Ca; pH 6.2), Liquozyme® X (0.4 kg/t DS; 10 ppm Ca; pH 5.4), and Liquozyme Supra (0.4 kg/t DS; 10
ppm Ca; pH 5.4).

Color formation
Liquozyme X and the Liquozyme Supra products can be applied at low pH. This
makes it possible to reduce color formation by e.g. Maillard reactions that take
place much faster at higher pH.

Savings on chemicals and ion exchange costs

Compared with Termamyl 120 L, Type L, Liquozyme X and the Liquozyme
Supra products can give significant cost savings. The calcium dosage can be
reduced or, in most cases, completely eliminated because Liquozyme X and the
Liquozyme Supra products are stable even at 5 ppm calcium.

When applying Liquozyme X or the Liquozyme Supra products, the amount of

pH adjustment chemicals required to increase the pH of starch slurry and to
decrease the pH of starch hydrolysate prior to saccharification can be reduced
because it is possible to run liquefaction at lower pH values.
When Liquozyme X or the Liquozyme Supra products are applied in liquefaction
at low calcium concentration and low pH, the reduced chemical addition results
in major savings on ion exchange costs. These savings give better overall
process economy because the savings are normally greater than the additional
enzyme cost.

Savings consist of reduced regeneration chemical consumption, lower resin

consumption due to longer operation cycles, and savings on water
consumption and wastewater treatment.

Please contact Novozymes' Customer Solutions for a calculation of possible

savings on production costs.
Usage
Application/process type
Termamyl 120 L, Type L, Termamyl 2X, Liquozyme X, and the Liquozyme Supra
products are designed to perform well in primary and secondary liquefaction of
starch for sweetener production.

In most applications, the liquefaction alpha-amylase should be added in a single

dosage to the starch slurry after pH adjustment and possible calcium salt
addition, but prior to steam jetting for the low-temperature process (105–
108 °C/221–226 °F). For the high-temperature process, the alpha-amylase is
added after flashing and pH adjustment. The retention time in the starch slurry
tank should be minimized to make full use of the enzyme activity.

Typical liquefaction processes

The recommended parameters for the low-temperature process are as follows:

• Enzyme added to starch slurry tank

• Primary liquefaction: 105–108 °C (221–226 °F) for 5–10 minutes depending
on raw material
• Flashing to vacuum
• Secondary liquefaction: 90–120 minutes at 95 °C (203 °F)

The recommended parameters for the high-temperature pasting process are as

follows:

• Primary liquefaction (no enzyme): 130–160 °C (266–320 °F)

• Flashing to atmosphere
• pH adjustment
• Enzyme addition
• Secondary liquefaction: 90–120 minutes at 98 °C (209 °F)

Dosage
The dosage requirement for liquefaction alpha-amylase depends on the process
conditions. The applied pH, DS, calcium, temperatures, and reaction times are
very important. However, the process equipment design also plays a significant
role. Primary liquefaction reactors normally consist of tubular coils and/or
mixed, multistage reactors. Secondary liquefaction reactors normally consist of
a number of reactors (in parallel or in series), with or without agitation.

For both primary and secondary liquefaction, it is important to design the

reactor system in such a way that a relatively narrow residence time distribution
is achieved.

The design of reactors in liquefaction determines the target DE. With a tubular
coil for primary liquefaction and 12–16 agitated tanks in series for secondary
liquefaction, a target DE of 8–10 is possible. For a smaller number of reactors, a
higher target DE is necessary.

Use the graphs below to establish the necessary dosage of liquefaction alpha-
amylase. Start by defining your target DE and desired liquefaction pH.
The target DE should normally be as low as possible so that the risk of
retrogradation is negligible.
The liquefaction pH should be as low as possible, considering the overall
economy of the process. Lower pH gives lower chemical and operation costs,
but requires a somewhat higher enzyme dosage.

Termamyl 120 L, Type L and Termamyl 2X perform well in the pH range 6.0–
6.4 but can be applied to pH levels as low as 5.8 (see Fig. 3).

pH 6.0 pH 6.2 pH 6.4

0,8

0,75
Dosage kg/t DS starch

0,7

0,65

0,6

0,55

0,5

0,45

0,4
9 10 11 12 13 14 15 16

DE (Fehling)

Fig. 3. Required dosage of Termamyl® 120 L, Type L versus required target DE.
70 ppm calcium and120 minutes.

pH 5.2 pH 5.3 pH 5.4

pH 5.5 pH 5.6

0,55
Dosage kg/t DS starch

0,5

0,45

0,4

0,35

0,3
8 9 10 11 12 13 14 15 16 17

DE (Fehling)
®
Fig. 4. Required dosage of Liquozyme X versus required target DE.
10 ppm calcium and 120 minutes.

Liquozyme X, Liquozyme Supra and Liquozyme Supra 2.2X perform well in the
pH range 5.2-5.6 but can be applied in the 4.9-5.7 range.
 pH 5.2 pH 5.3 pH 5.4
pH 5.5 pH 5.6

0,52

Dosage kg/t DS starch

0,47

0,42

0,37

0,32

0,27

0,22
7 8 9 10 11 12 13 14 15

DE (Fehling)

Fig. 5. Liquozyme® Supra dosage versus target DE at different pH values, at 10 ppm calcium, and
120 minutes. Required dosage of Liquozyme Supra versus required target DE.

Calcium level
The free calcium concentration is not important for Liquozyme X and the
Liquozyme Supra products. Only 5 ppm is necessary to stabilize the enzyme
during liquefaction. In most cases, this level of free calcium is readily supplied
by the starch raw material and/or the process water.

When the calcium concentration is increased, the enzyme becomes slightly

more stable during primary and secondary liquefaction. With liquefaction
conditions of 105 °C (221 °F) for 5 minutes followed by 95 °C (203 °F) for 120
minutes, DE is approximately 0.3% higher at 35 ppm compared with 10 ppm
calcium. For Termamyl 120 L, Type L and Termamyl 2X, the recommended level
of free calcium is 40–70 ppm.

Conductivity
Liquefaction alpha-amylases can be applied at specific conductivities as low as
300–400 μS/cm without any influence on DE development. At lower specific
conductivities, the dosage must be increased. It should be noted that when the
pH of the starch slurry is lowered and when calcium addition is reduced, the
conductivity of the slurry will also decrease.

Activity and stability

All six alpha-amylases are thermostable and suitable for normal liquefaction
conditions with primary liquefaction at 105–110 °C (221–230 °F) for 5–10
minutes followed by secondary liquefaction at 95 °C (203 °F) for 90–120
minutes. The relative performance of the liquefaction alpha-amylase versus
primary liquefaction temperature and pH is shown in Figures 6 and 7.
 Liquozyme® X Liquozyme Supra Termamyl® 120 L

110
Relative performance 100
90
80
70
60
50
40
30
20
100 105 110 115
(201ºF) (221ºF) (230ºF) (239ºF)
Temperature, ºC

Fig. 6. Relative performance of liquefaction alpha-amylases at various primary liquefaction temperatures.

Liquozyme® X Liquozyme Supra Termamyl® 120 L

100
90
Relative performance

80
70
60
50
40
30
20
10
0
4,8 5,0 5,2 5,4 5,6 5,8 6,0 6,2 6,4

pH

Fig. 7. Relative performance of liquefaction alpha-amylases versus pH in primary liquefaction.

The enzymes can be inactivated by lowering the pH and holding at 95 °C

(203 °F).

Depending on the required degree of inactivation, the enzyme should be held

for 5–10 half-lives.
 Enzyme pH Calcium T½ (min)
(ppm)
Termamyl® 120 L, Type 4.3 18 ~1
L
Termamyl 2X
Liquozyme® X 3.8 18 ~2
Liquozyme Supra 3.8 10 ~4
Liquozyme Supra 2.2X
Liquozyme Supra 2.8X
Table 2. Enzyme half-lives at 95 °C (203 °F).

For Termamyl 120 L, Type L and Termamyl 2X, inactivation prior to

saccharification is achieved by adjusting the pH to 4.3 and holding in a tank or
coil for 5–10 minutes at 95 °C (203 °F). Liquozyme X and the Liquozyme Supra
products are much more stable, but inactivation before saccharification is
normally not necessary. For the production of certain specialty syrups (e.g.,
80+% maltose) or for the complete inactivation of liquefaction enzyme in the
final products, we recommend carrying out thermal inactivation by increasing
the temperature to 115–135 °C (239–275 °F) for a short time.

Production of maltodextrin
For the production of low-DE maltodextrin, BAN 480 L may be applied. BAN
480 L is an alpha-amylase, which performs well at temperatures up to 85 °C
and pH around 6.0 and requires 50–100 ppm calcium for stabilization.

BAN is typically used in secondary liquefaction, where its dextrin breakdown

pattern and ease of inactivation are beneficial.

Production of glucose and maltose syrups

When BAN 480 L or Fungamyl® 800 L are used to produce a specific sugar
composition during saccharification of glucose or maltose syrups, it should be
ensured that the calcium concentration is sufficient (50–100 ppm).

Thus, if the liquefaction process is run with one of the Liquozyme Supra
products or Liquozyme X at low calcium concentration, it may be necessary to
add calcium for saccharification.

Storage in application
Recommended storage conditions are 0–25 °C (32–77 °F) in sealed packaging,
protected from the sun. The product has been formulated for optimal stability.
However, enzymes gradually lose activity over time. Extended storage and/or
adverse conditions such as higher temperature may lead to a higher dosage
requirement.
 Safety, handling, and storage
Safety, handling, and storage guidelines are provided with all products.

Novozymes A/S For more information Laws, regulations, and/or third party rights may prevent
customers from importing, using, processing, and/or
Krogshoejvej 36 and addresses of reselling the products described herein in a given
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the customer and Novozymes to such effect, this
Denmark please see document does not constitute a representation or
www.novozymes.com warranty of any kind and is subject to change without
further notice.
Tel. +45 4446 0000 starch@novozymes.com ® Novozymes A/S · No. 2003-33447-06
Fax +45 4446 9999