INTRODUCTION

Fruit juice is 100% pure juice made from the flesh of fresh fruit or from whole fruit,
depending on the type used. It is not permitted to add sugars, preservatives, sweeteners,
flavourings and colourings to fruit juice. One of the most common fruit products is fruit
juices. Fruit juice can take many forms, including a natural style cloudy product, a nectar type
product containing suspended solids, a fully clarified juice, juice concentrate and fruit drinks.
If the juice is to be clarified further or concentrated after extraction, treatment with pectinase
may be required. Pectinase is an enzyme used commercially to aid in extracting juice from
fruit (Mark, 2021). Juice is composed of water, soluble solids, aroma and flavour compounds,
vitamins and minerals, pectic substances, protein and fats. Pectinase treatment not only
improves the juice quality by lowering its viscosity and turbidity but also it improves its
colour. Enzyme treatment makes the juice clear and transparent by removing the unwanted
haze due to pectin fiber as well as improves the visibility by preventing the darkening of juice
and providing the characteristics colour of the juice. Different fruit juices need different type
of enzyme treatment depending upon the consumer’s acceptance. To obtain the desired
product, process parameters are needed to be optimized such as enzyme concentration,
enzyme treatment duration, incubation temperature, and ph of solution. (Jyoti, 2018)

Enzymes are an integral component of modern fruit juice manufacturing and are
highly suitable for optimizing processes. Their main purposes are increase extraction of juice
from raw material, increase processing efficiency (pressing, solid settling or removal), and
generate a final product that is clear and visually attractive. Juice extraction can be done by
using various mechanical processes, which may be achieved through diffusion extraction,
decanter centrifuge, screw type juice extractor, fruit pulper and by different types of presses.
Enzymatic treatment prior to mechanical extraction significantly improves juice recovery
compared to any other extraction process. Enzymatic hydrolysis of the cell walls increases
the extraction yield, reducing sugars, soluble dry matter content and galacturonic acid content
and titrable acidity of the products.

OBJECTIVES

1. To describe the role of enzyme (pectinase) in extracting and clarifying fruit juice
2. To explain the yield , quality and acceptance of clarified and non clarified juice from
watermelon
APPARATUS AND MATERIALS

APPARATUS MATERIALS

Refractometer Fully ripe watermelon

Induction cooker Pectinase enzyme

UV-Vis Spectrophotometer

Water bath

Blender

Weighing balance

pH meter

Colorimeter
METHODOLOGY

150 g of the flesh was weighed and added into 300ml of water

The sample was blended using a blender

The puree was pasteurized at 90C for 10 minutes

The extract was cooled to about 38.5C using water bath

The extracted juice was treated with different concentration (%v/v) of pectinase
enzyme (0.00%, 0.025%, 0.05%, 0.075%, 0.1%) at 38.5C for one hour in a
water bath

The suspension was heated at 90C for 10 minutes

The juice was filtered using muslin cloth and been weighed.

The filtered juice was collected for analysis

RESULTS

TOTAL
ENZYME YIELD SOLUBLE CLARITY
PH COLOUR
CONCENTRATION (%) SOLIDS AT 660NM
(BRIX)

L 3.67
0.000 % 75.00 6.12 6.05 a 3.72 0.281
b 3.79
L 5.98
0.025 % 78.41 5.70 4.70 a 3.32 0.212
b 3.63
L 15.03
0.050 % 69.77 5.69 6.18 a 3.85 0.511
b 5.11
L 6.86
0.075 % 77.28 5.71 4.56 a 3.52 0.351
b 2.73
L 5.79
0.100 % 79.07 5.80 5.91 a 3.35 0.263
b 3.58

CALCULATION
% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)

0.00 % OF ENZYME CONCENTRATION

% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)
= 330ml x 100%
440ml
= 75%
0.025 % OF ENZYME CONCENTRATION
% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)
= 345ml x 100%
440ml
= 78.41%

0.050 % OF ENZYME CONCENTRATION

% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)
= 300ml x 100%
430ml
= 69.77%

0.075 % OF ENZYME CONCENTRATION

% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)
= 330ml x 100%
427ml
= 77.28%

0.100 % OF ENZYME CONCENTRATION

% Yield = Amount of juice after treatment (ml)
Amount of homogenized juice (ml)
= 340ml x 100%
430ml
= 79.07%
DISCUSSION

Clarification is a process by which the semi stable emulsion of colloidal plant

carbohydrates that support the insoluble cloud material of a freshly pressed juice is
“broken” such that the viscosity is dropped and the opacity of the cloudy juice is
changed to an open splotchy look. Pectinases are a group of an enzyme capable of
hydrolyzing pectin substances or degrading molecules that are found in the middle
lamella and primary cell walls of plants. The enzymatic process is claimed to offer a
number of advantages over mechanical-thermal comminution of several fruit pulps. In
particular, the use of pectinases has been an integral part of modern fruit processing
technology involving treatment of fruit masses they not only facilitate easy pressing
and increase in juice recovery but also ensure the highest possible quality of end
products.

From the result obtained, 0.1% of pectinase enzyme have higher percentage of
yield compared to other concentration of pectinase enzyme. This shows that pectinase
enzyme not only help in softening the plant tissue but also lead to the release of cell
contents that recovered with high yield. Fruit contains pectin and other
polysaccharides so it may lead to fouling during filtration through membrane.
Enzymatic treatment leads to degradation of pectin. Enzymatically clarified juice
resulted in viscosity reduction and cluster formation, which facilitates separation
through centrifugation or filtration.

The pH value of juice decreases with increase in enzyme concentration. pH

can be measured by digital pH meter. Results shows that 0.1% have low pH compared
to 0.0 % of pectinase, this indicate that the juice will become more acidic when use
the pectinase enzyme. The concentration of H+ increase in juice when pectinase was
added. The brix value can be determined by measuring the refractive index with a
hand held refractometer. Results shows that 0.05% have high total soluble solids
compared to other concentrations. The increase in Total Soluble Solid is related to
greater degree of tissue breakdown, releasing more compounds such as sugars, which
contribute to soluble solids. For the colour, 0.05% shows higher value of “L” which
is +15.03. This shows the the 0.05% of pectinase contained juice have brighter
compared to other concentration. For the value of “a” and “b” all the concentration
shows a positive value which is red for “a” and yellow for “b”.
 Enzymatic treatment leads to increase the clarity of juice. Juice clarity can be
determined in terms of absorbance and transmittance at 660 nm using UV visible
Spectrophotometer. Increase in enzymatic concentration increase the rate of
clarification by exposing part of the positively charged protein beneath thus reducing
electrostatic repulsion between cloud particles which caused these particles to
aggregate into larger particles and eventually settled out. Pectinase concentration of
0.1% have obtained 0.263 absorbance which shows that the lowabsorbance values at
highest enzyme concentration, where lower absorbance indicates a clearer juice is
being produced. Hence, 0.1 % of pectinase shows low absorbance value compared to
0.0% of pectinase but 0.05% shows 0.511 absorbance value which is very high. This
can assume that there are few error had occur such as they placed the juice in UV-Vis
Spectrophotometer before the juice is cool, the cuvette placed was not handle with
precautions step and an improper techniques was used in filtration. In summary, all
the precaution steps should be follow in order to produce an accurate result.
CONCLUSION

The use of pectinases has been an integral part of the modern fruit processing
technology involving treatment of fruit masses. Enzymes in combination claim to
increase juice recovery, Total Soluble Solid, clarity and decrease viscosity and
decrease pH. Cellulytic and pectolytic enzymes mixtures are having wide application
to enhance pulp liquefaction and provide a higher yield of juice with high soluble
solids content. As a result, the juice presents higher clarity, as well as more
concentrated flavor and color. In a nutshell, 0.1% pectinase enzyme is an optimum
concentration to juice clarification where higher the concentration, higher the yield
produced.
QUESTIONS

1. What is the effect of varying the enzyme concentration?

Enzymes have this ability to speed up reactions by lowering the activation energy, the
energy needed to break the bonds of the reactants. To determine the optimum
concentration of pectinase for juice clarification. Different concentration of enzyme
will give different result in the aspect of yield percentage, pH, total soluble solids,
colour and clarity of juice.

2. Explain what will happen if amylase is added in combination with pectinase.

Amylase is an enzyme that catalyses the breakdown of starch into sugars. Amylase
used clarification of fruit juice. Amylase will increases the juice yield and reduces the
processing time, but also improves the extraction of valuable fruit components, and
after the juice extraction, whereby amylase are used for its clarification, thereby
lowering the viscosity of fruit juice prior to concentration and increasing the filtration
rate and stability of the final product. Moreover, amylase will increase the sweetness
of juice.
REFERENCES

Abbes, F., Bouaziz, M., Blecker, C., Masmoudi, M., Attia, H. and Besbes, S. (2011).
Date syrup: Effect of hydrolytic enzymes (pectinase/cellulase) on physicochemical
characteristics, sensory and functional properties. Food Sci. Technol. 44: 1827–1834

Mark R. 2021. Fruit Processing. Retrieved From

https://www.britannica.com/topic/fruit-processing/Fruit-preserves-jams-and-jellies

Science Buddies Staff. (2020, January 12). A Juicy Project: Extracting Apple Juice
with Pectinase. Retrieved from https://www.sciencebuddies.org/science-fair-
projects/project-ideas/BioChem_p010/biotechnology-techniques/extracting-apple-
juice-with-pectinase

Jyoti Singh Jadaun. 7th march 2018. Pectinase : A useful Tool in Fruit Processing
Industries. Retrieved from
https://juniperpublishers.com/nfsij/pdf/NFSIJ.MS.ID.555673.pdf

Joshi, A. A., Kshirsagar, R. B. and Sawate, A. R. (2012). Studies on standardization

of enzyme concentration and process for extraction of tamarind pulp, variety Ajanta. J.
Food Proc. Technol. 3(2):1–3

Kaur, S., Sarkar, B. C. and Sharma, H. K. (2009). Optimization of enzymatic

hydrolysis pretreatment conditions for enhanced juice recovery from guava fruit using
response surface methodology. Food Bioprocess Technol. 2:96–100.

Yusuf S, Ibrahim N. Quality of sour soup juice after pectinase enzyme treatment.
Food Chem. 1994;51:83–88. doi: 10.1016/0308-8146(94)90052-3