Food background notes

Enzymes and carrot juicing

Carrots are a nutritious vegetable. They contain a good

supply of the precursors of vitamin A (retinol), with the
main ones being -carotene and -carotene. Carrots
also supply dietary fibre, minerals and a range of other
vitamins. However there are challenges in the
production of juice related to the enzymes naturally
present within the carrot after harvesting. [1]

The processing of carrot juice The primary

problem is that, as soon as we disrupt the cellular
structure of the plant tissue, compartmentalisation is
lost. This affects cell walls, cell membranes and well as
the membranes enclosing sub-cellular structures
including vacuoles, nucleii and chloroplasts. This
important as these barriers usually are able to keep
various components so that these are unable to interact
or undergo chemical reactions

Blanching of carrots Some of the options which

have been studied are the use of steam, boiling water
and microwave treatment [4]. The results of
comparative trials showed that microwave was not
effective for the uniform heating of the carrots, partly
due to their tapered shape. Steam and boiling treatments
were quite effective approaches to blanching. The study
was extended to a comparison of four common genetic
varieties of carrots [4].
Blanching and the inactivation of enzymes
When the three enzymes were assayed during steam
blanching, in all cases it was confirmed that higher
temperatures resulted in more rapid inactivation. As an
example of the pattern, the results for PPO for one
particular variety of carrots (RHC 100) are presented in
Figure 1.

As with many living tissues, as long as there is no

juicing or grinding, the enzymes present are effectively
idle, because they do not have access to their
substrates. However upon juicing, as a result of the loss
of compartmentalisation, various enzymes naturally
present in the carrots will be able to mix freely with
their substrates which are also commonly present
within the cells of carrots. The resultant reactions are
undesirable as they influence quality of food products
and so the enzymes are referred to as being
deteriorative.

Deteriorative enzymes in carrots There are

three deteriorative enzymes found to be a serious
problem in the processing of carrots:
1. Polyphenol oxidase (PPO) which is responsible for
browning reactions. There are a number of distinct
enzymes which are included as PPO and have
similar effects during processing. These vary in
their specificity for particular substrates [2].
2. Peroxidase (PO, EC 1.11.1.7) which is involved
with oxidation reactions and the main substrate is
hydrogen peroxide (H2O2) [3]. This enzyme is
known to cause off-flavours in various foods
3. Pectin methyl esterase (PME, EC 3.1.1.11) which is
one of the enzymes needed for the hydrolysis of
pectin molecules [3]. These polysaccharides are a
major component form the basic structure of the
cell walls of carrots.
Each of these enzymes impacts directly on the quality
and shelf-life of carrot juice. In order to overcome
these deteriorative effects we will need to blanch the
carrots before any disruption of the carrot tissues.
The term blanching refers to any brief treatment
applied in order to minimise deteriorative changes.
These might be applied immediately post-harvest or
alternatively just prior to the commencement of
processing of a vegetable or other ingredient.

Figure 1

The inactivation of PPO in carrots during

steam blanching at various temperatures
(RHC is variety Red Hot Carotene 100)

Thermal inactivation was assessed over a range of

temperatures and our research showed that the three
enzymes are inactivated at different temperatures. PPO
was found to be the least stable and pectinesterase the
most stable for each of the four varieties studied. In
most cases effective inactivation was achieved within 2
minutes at 85C (as in Figure 1) [4].

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continued.........

Food background notes:

enzymes and carrot juice

In addition there was some variation between the results

for when each of the enzymes was compared for the
four varieties. The enzymes in one (Top Pak) showed
greater thermal stability. In this case, pectinesterase
required treatment at 90C to ensure rapid inactivation.
Further analyses showed that there was uneven
distribution of the enzymes in the core and outermost
layers of the carrot root (Table 1). On the other hand no
variation was found between the two ends of the root
(tip and stem end, data not shown) [4].

Selection of treatment conditions In processing

situations it is generally necessary to blanch using
conditions which result in inactivation of the most
stable of the enzymes. At the same time it is important
to minimise the treatment so that the texture and flavour
of the resultant product is not modified too much. For
carrots this means that shorter periods of heating reduce
the likelihood that there are caramelisation reactions
influencing flavour. Consumers do not want a juice
product to have a cooked taste. Our studies also
indicated that there were differences in the effects of
blanching for a series of different genetic varieties of
carrots
Indicator enzymes In processing it is common to
use one enzyme as an indicator. Thus, if we measure
that enzyme and find that it has no activity, we can be
sure that other deteriorative enzymes have also been
inactivated. For carrots, it was concluded that
pectinesterase should be used as the indicator enzyme
in the assessment of blanching sufficiency.

Table 1

page 2 of 2
The relative distribution of deteriorative
enzymes in carrots (expressed as a
percentage of total activity, for variety
RHC100) [4]

Enzyme

Superficial

Core

Peroxidase

72.8

27.2

PPO

100

Pectinesterase

53.4

46.6

So the combined commercial enzymes are added at

lower levels and allowed to act on the juice prior to
centrifugation/clarification filtration treatments. Note
that the commercial enzymes are probably inactivated
by subsequent steps in the procedure because there is a
concentration step applied to reduce the bulk and hence
the cost of transportation of the juice.

Conclusion The research showed that the one

particular enzyme can be both deteriorative as well as
beneficial. The fundament principle here is that the
processor must be in control of what is happening at all
times. This includes, for example, temperatures, how
times, as well as the relative amounts of the two
commercial enzyme preparations .
References and further reading
[1] Vora HM, Kyle WSA, Small DM. 1998.
Composition and juicing potential of Australian
varieties of carrots. Food Aust 50(1):24-26.

Post blanching treatment of carrots Once the

carrots are blanched, cooled and passed through a juicer
we have a thick solution which is quite cloudy. Some
form of filtration/centrifugation step is needed in
processing so that the final product is relatively uniform
in appearance and there is as little sediment as possible.
In the research on carrots it was found that the addition
of two commercial enzyme preparations to the juice had
a number of benefits.
The two enzymes had commercial names of Rohament
(given by the manufacturer) but the actual active
enzymes present in these were PME (the same as
referred to above as a deteriorative enzyme) and
another called polygalacturonase. This latter enzyme
also acts to break down pectin molecules, in this case
by breaking the links between adjacent monosaccharide
units in the backbone of the pectin molecule.

[2] Ramrez EC, Whitaker JR, Virador VM. 2002.

Polyphenol Oxidase. In: Whitaker JR, Voragen AGJ,
Wong DWS, editors. Handbook of food
enzymology. New York: Marcel Dekker. 39 p1-15.
[3] Nomenclature Committee of the International Union
of Biochemistry and Molecular Biology. 2011.
Enzyme nomenclature. [Online. Internet.] Available
from: http://www.chem.qmul.ac.uk/iubmb/enzyme/.
[4] Vora HM, Kyle WSA, Small DM. 1999. The
activity, location and thermal inactivation of three
deteriorative enzymes from Australian carrot
(Daucus carota L) varieties. J Sci Food Agric
79(8):1129-35.
[5] Vora HM, Kyle WSA, Small DM. 1999. The
application of commercial enzyme preparations in
the production of juice from Australian carrots. Food
Aust 51(6):122-23.

This provides two advantages, the first being greatly

enhanced yields of juice because the cell contents are
more readily released from within the cell structures.
The second was higher quality juice because there is
more sugar (enhancing flavour) and carotenes (good
nutritionally but more importantly giving a brighter
colour and hence visual appeal to the product.

Prepared by Assoc Prof Darryl M Small

send feedback or questions to <darryl.small@rmit.edu.au >