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EFFECT OF CALCIUM PROPIONATE ON THE INHIBITION OF FUNGAL GROWTH

IN BAKERY PRODUCTS

Article · January 2015

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Volume – 1; Issue - 3; Year – 2015; Page: 273 – 279

Indo – Asian Journal of Multidisciplinary Research (IAJMR)

ISSN: 2454-1370

EFFECT OF CALCIUM PROPIONATE ON THE INHIBITION OF FUNGAL

GROWTH IN BAKERY PRODUCTS
D. Prem Kumar1*, M. Jayanthi1, P. Saranraj2 and S. Kavi Karunya1
1
Department of Microbiology, Annamalai University, Annamalai Nagar – 608 002, Tamil Nadu, India.
2
Assistant Professor of Microbiology, Department of Biochemistry, Sacred Heart College (Autonomous),
Tirupattur – 635 601, Tamil Nadu, India.
Abstract
Bakery products are valuable source of nutrients. Bakery products are considered as a source of
carbohydrates because starch is the main chemical constituent. Bakery products are subjected to spoilage
problems. Mould spoilage is a serious and costly problem for bakeries. The stability of bakery products
against the attack by fungi is mainly due to preservatives. Propionates were effective in controlling mold growth
on the surface of bakery products. In the present study, the effect of chemical preservative Calcium propionate
against the bakery food spoilage fungi was tested in different concentrations, pH and temperature respectively.
Among the various concentrations 0.25%, pH 7.5 and 20 °C was highly effective in the control of Rhizopus
stolonifer, Aspergillus niger, Penicillium chrysogenum and Mucor sp.

Key words: Food spoilage fungi, pH, Temperature and Calcium propionates.

1. Introduction
Bakery products are an important source of life of bakery products. It plays a decisive role
nutrients viz., energy, protein, iron, calcium and when molds compete with bacteria to spoil high
several vitamins. Fibers enrichment of several moisture foods (Ponte et al., 1993). Preservatives
bakery products has recently been tested using an are most commonly used to control mold growth
ingredient containing 95 per cent short chain in baked goods (CFR, 1990). Propionic acid, an
fructo-oligosaccharides (Hocking, 1998; Geetha et aminocarbolic acid (CH3CH2-COOH), is a
al., 2012; Jageethadevi et al., 2012). naturally occurring organic acid and is an oily
liquid with a slightly pungent, disagreeable rancid
Physical spoilage of bakery products usually odor. Its salts are white, free-flowing powders
involves moisture loss or gain resulting in a loss of with a slight cheese like flavor (Doores, 1993).
texture or mold growth. Microbiological spoilage Propionates were selected on the basis that higher
is often the major factors limiting the shelf life of MW fatty acids had a higher antimicrobial effect.
bakery products. Spoilage from microbial growth This acid or its salt can be used to prevent the
causes economic loss for both manufacturers and bacterial spoilage of bread known as rope caused
consumer (Hickey, 1998). This problem usually by certain Bacillus sp. (Legan, 1993). Several
occurs in the summer season when the climate is studies have also reported the effects of propionic
warm and humid (Smith, 1993). Physical factors acid and its salt on mold growth. Concentrations
are the important factor governing mold free shelf of propionate ranging from 8 to 12 per cent have
been reported effective in controlling mold growth
*Corresponding author: D. Prem Kumar on the surface of bakery products (Doores, 1993).
E-mail: [email protected]
Received: 25.06.2015; Revised: 20.07.2015;
Accepted: 18.08.2015.

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Prem Kumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 1(3): 273 – 279 274

2. Materials and methods Sabouraud’s dextrose broth was prepared

and distributed at 50 ml quantities in 100 ml
Collection of bakery foods
Erlenmeyer flasks in different concentration. The
Five different spoiled bakery foods were
calcium propionate (0.0%, 0.5%, 0.10%, 0.15%,
collected from the bakery in Chidambaram, Tamil
0.20%, and 0.25%) were prepared and added to
Nadu, India. The collected spoiled bakery foods
the broth and inoculated with 1 ml of fungal
were;
inoculums separately and incubated at room
a) Wheat bread. d) Chocolate cake. temperature for 3 days. After the incubation period
b) Sponge cake. e) Putting cake. the growths were measured at 420 nm using
c) Plum cake. Spectrophotometer.

Isolation and identification of fungi from Effect of calcium propionate at different pH on

bakery foods the inhibition of fungal growth
The samples were processed for the Sabouraud’s dextrose broth was prepared
estimation of fungal population by dilution plating and distributed at 50ml quantities in 100 ml
method by suspending 1g of each sample in 100 Erlenmeyer flasks in different concentration. The
ml sterile water in conical flask. The flask was preservative (0.0%, 0.5%, 0.10%, 0.15%, 0.20%,
kept in mechanical shaker for 5 minutes to prepare and 0.25%) were added to the broth and the pH
the suspension. One ml of aliquots was pipetted was adjusted to various level from 4.0, 4.5, 5.0,
out to 10 ml sterile water and dispersed uniformly 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0 in each flask by
by shaking. One ml of sample was transferred adding 0.1 N NaOH or 0.1N HCl or and pH in
repeatedly till a dilution of 10-4 or 1/10,000 was each broth was tested with the help of glass
obtained. Aliquots of 1 ml of the serial dilution electrode pH meter. The flasks were inoculated
10-3 and 10-4 were pipetted out to sterile with 1 ml of fungal inoculums separately and
petridishes and the plates were poured with SDA incubated at room temperature for 3 days. After
medium. The medium was allowed to set and the incubation period the growths were measured
incubated in inverted position. All plates were at 420 nm using Spectrophotometer.
incubated at room temperature for 3 days. The Effect of calcium propionate at different
colonies were counted and expressed per gram of temperature on the inhibition of fungal growth
bakery products. Sabouraud’s dextrose broth was prepared
The Lactophenol cotton blue mount was and distributed at 50 ml quantities in 100 ml
especially used to identify the fungus. In this a Erlenmeyer flasks in different concentration. The
drop of Lactophenol cotton blue was placed on a preservative (0.0%, 0.5%, 0.10%, 0.15%, 0.20%,
clean slide. With the help of the teasing needle and 0.25%) were prepared and added to the broth
small portion of the colony was picked and it was and inoculated with 1 ml of fungal inoculums
spread on Lactophenol cotton blue by using separately and incubated for 3 days at different
another teasing needle. Then, the cover slip was temperature viz., 10 °C, 15°C, 20 °C, 30 °C, 35
placed over that without any air bubbles. Then, the °C, 40 °C, and 45 °C in an incubator. After the
slide was examined under 45x objective lens. incubation period the growths were measured at
420 nm using Spectrophotometer.
Effect of calcium propionate on the inhibition
of fungal growth 3. Results and Discussion
The inhibitory effects of different Spoilage by mold is one of the major
concentrations of calcium propionate on the fungal problems in bakery food and these results will
growth were studied in the laboratory (Grundy, cause more economic losses. In this present study,
1996). five different bakery products viz., wheat bread,

© 2015 Published by JPS Scientific Publications Ltd. All rights reserved

Prem Kumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 1(3): 273 – 279 275

sponge cake, plum cake, chocolate cake and propionate was effective in the control of bakery
putting cake were collected from the bakery in product spoilage fungi (Grundy, 1996).
Chidambaram. The isolated cultures were
identified as Rhizopus stolonifer, Aspergillus In the present study, the effect of the chemical
niger, Penicillium chrysogenum and Mucor sp. preservative, Calcium propionates against bakery
food spoilage causing fungi was tested under different
Vytrasova et al. (2002) detected, isolated pH (4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0) and
and identified xerophilic fungi Eurotium the results were showed in Fig - 2, Fig - 3, Fig - 4
amstelodami, Eurotium chevalieri, Eurotium and Fig - 5. The preservative were used in different
herbariorum, Eurotium rubrum and Wallemia concentrations viz., 0.0 %, 0.5 %, 0.10 %, 0.15 %,
sebi. The resistance of these fungi against elevated 0.20 % and 0.25 %. The growth of the fungi Rhizopus
temperature and preserving agents was stolonifer (0.079 at 420 nm), Aspergillus niger (0.045
investigated. It was found that Eurotium sp. were at 420 nm), Penicillium chrysogenum (0.054 at 420
more resistant than Wallemia sebi. Preservation nm) was highly inhibited at pH 7.5 whereas, in Mucor
against xerophilic fungi was more effective with sp. (0.078 at 420 nm) was highest at pH 7.0. For all
the use of propionic acid than with potassium the fungal isolates, the growth was completely
sorbates. arrested at pH 4.0 and 8.0 (Doores, 1993).
In Propionates, 0.25% was highly effective Fig – 2: Effect of Calcium propionate at different
for the control of the growth of Rhizopus pH on the inhibition of Rhizopus stolonifer growth
stolonifer (0.182 at 420nm), Aspergillus niger
(0.189 at 420nm), Penicillium chrysogenum 0.45
(0.195 at 420nm) and Mucor sp. (0.177 at 420nm) 0.4
in Fig-1. 0.35
OD at 420 nm

0.3
Fig – 1: Effect of Calcium propionate on the 0.25
inhibition of fungal growth 0.2
0.15
0.35 0.1
0.3 0.05
0
OD at 420 nm

0.25
0.2 4 4.5 5 5.5 6 6.5 7 7.5 8
0.15 pH
0.1
0.05 0% 0.50% 0.10%
0
0.15% 0.20% 0.25%
0 0.5 0.1 0.15 0.2 0.25
Conc. of propionate (%)
The effect of chemical preservative
Calcium propionate against bakery food spoilage
R. stolanifer A. niger
fungi was tested under different temperature
P. chrysogenum Mucor sp. (10°C, 15°C, 20°C, 30°C, 35°C, 40°C, and 45°C)
and the results were showed in Fig - 6, Fig - 7, Fig
The growth of the Mucor sp. was highly - 8 and Fig - 9. The preservative were used in
controlled when compared to Rhizopus stolonifer, different concentrations viz., 0.0 %, 0.5 %, 0.10 %,
Aspergillus niger and Penicillium chrysogenum. 0.15 %, 0.20 % and 0.25 %. The growth of the
The results clearly showed that the Calcium fungi Rhizopus stolonifer (0.075 at 420 nm),
Penicillium chrysogenum (0.087 at 420 nm) and

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Prem Kumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 1(3): 273 – 279 276

Mucor sp. (0.031 at 420 nm) was highly inhibited Fig – 5: Effect of Calcium propionate at different pH
at 15 °C. Whereas, the growth of the fungi on the inhibition of Mucor sp. growth
Aspergillus niger (0.115 at 420 nm) was highest at
0.4
20 °C. For all the fungal isolates, the growth was

OD at 420 nm
completely inhibited at 10 °C, 40 °C and 45 °C. 0.3
0.2
Fig – 3: Effect of Calcium propionate at different
pH on the inhibition of Aspergillus niger growth 0.1
0
0.4
4 4.5 5 5.5 6 6.5 7 7.5 8
OD at 420 nm

0.3
pH
0.2
0.1 0% 0.50% 0.10%
0
0.15% 0.20% 0.25%
4 4.5 5 5.5 6 6.5 7 7.5 8
pH Fig – 6: Effect of Calcium propionate at different
0% 0.50% 0.10% temperature on the inhibition of Rhizopus stolonifer
growth
0.15% 0.20% 0.25%

0.45
0.4
Fig – 4: Effect of Calcium propionate at different
0.35
pH on the inhibition of Penicillium chrysogenum
OD at 420 nm

0.3
growth
0.25
0.2
0.4
0.15
OD at 420 nm

0.3 0.1
0.2 0.05
0
0.1
10 15 20 30 35 40 45
0
Temperature (C)
4 4.5 5 5.5 6 6.5 7 7.5 8
pH 0.0% 0.5% 0.10%
0.15% 0.20% 0.25%
0% 0.50% 0.10%
0.15% 0.20% 0.25%
Water activity, CO2, and their interaction
were the main factors significantly affecting
Guynot et al. (2003) used a sponge cake fungal growth. Water activity at levels of 0.80 to
analogue to study pH, water activity (a w) and 0.90 had a significant influence on fungal growth
carbon dioxide (CO2) levels on the growth of and determined the concentration of CO2 needed
seven fungal species commonly causing bakery to prevent cake analogue spoilage. At an a w level
product spoilage (Eurotium amstelodami, of 0.85, lag phases increased two fold when the
Eurotium herbariorum, Eurotium repens, level of CO2 in the headspace increased from 0 to
Eurotium rubrum, Aspergillus niger, Aspergillus 70 per cent. In general, no fungal growth was
flavus, and Penicillium corylophilum). observed for upto 28 days of incubation at 25 C

© 2015 Published by JPS Scientific Publications Ltd. All rights reserved

Prem Kumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 1(3): 273 – 279 277

when samples were packaged with 100% CO2, the solid salt than the corrosive liquid acid (Seiler,
regardless of the aw level. 1994). Sodium, potassium, and calcium propionate
are the most widely used antimicrobial additives
Fig – 7: Effect of Calcium propionate at in the baking industry. They are active against
different temperature on the inhibition of molds and have little effect against bacteria
Aspergillus niger growth (Smith, 1993). Both calcium propionate and
sodium propionate have been affirmed as GRAS
0.4 (Grundy, 1996).
0.35
0.3 Fig-8. Effect of propionate at different
OD at 420 nm

0.25 temperature on the inhibition of Penicillium

0.2 chrysogenum growth
0.15
0.1 0.4
0.05 0.35
0.3

OD at 420 nm
0
0.25
10 15 20 30 35 40 45
0.2
Temperature (C) 0.15
0.1
0.0% 0.5% 0.10% 0.05
0
0.15% 0.20% 0.25%
10 15 20 30 35 40 45

Sauer and Burrughs (1993) observed that Temperature (C)

the mold free shelf life of corn was extended for 1
week by 0.5-l. per cent calcium propionate. In the 0.0% 0.5% 0.10%
United States, preservation of bread for a few days 0.15% 0.20% 0.25%
is generally accomplished by the addition of
sodium or calcium propionate. However, some Fig-9. Effect of propionate at different
species of PencilIium are resistant and can grow in temperature on the inhibition of Mucor sp. growth
media containing 5 per cent propionic acid
(Doores, 1993). The advantage of using 0.35
propionate is that it has little effect on yeast hence, 0.3
OD at 420 nm

propionates can be added to bread dough to 0.25

prevent rope bacteria and mold growth without 0.2
interfering with leavening (Grundy, 1996). 0.15
Moreover, propionic acid and propionate are used 0.1
at levels of 0.1 and 0.2 per cent by flour weight, 0.05
respectively (Seiler, 1994; Saranraj and Geetha, 0
2012; Saranraj and Naidu, 2014). 10 15 20 30 35 40 45

At these levels, the mold free shelf life can Temperature (C)
be increased by 1-2 d. Compared to sodium
benzoate; propionates are generally more active 0.0% 0.5% 0.10%
against mold (King, 1981). However, propionic 0.15% 0.20% 0.25%
acid caused undesirable odors and flavors in baked
goods. Therefore, the bakers prefer to use solid
calcium propionate because it is easier to handle

© 2015 Published by JPS Scientific Publications Ltd. All rights reserved

Prem Kumar / Indo – Asian Journal of Multidisciplinary Research (IAJMR), 1(3): 273 – 279 278

Marin et al. (2002) tested the use of weak- C.C. (Ed), Food preservation by moisture
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