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1 1 17.1 PDF
1 1 17.1 PDF
ISSN: 2455-4227
www.newresearchjournal.com/science
Volume 1; Issue 1; January 2016; Page No. 05-08
A comparative study on Lactic acid production from Canistel fruit using Lactobacillus species
1
Mridul Umesh, 1 Thazeem B, 2 Vikas OV
1
Research Scholar, Department of Microbial Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, India.
2
Research Chemist, RSAS Laboratories, Ajman, United Arab Emirates.
Abstract
The high production cost involved in the chemical synthesis of Lactic acid along with its ever increasing demand due to wide
spread application calls for new approaches to screen and produce LA from cheap and inexpensive raw material. The main focus
of the research work is to produce LA from an under-utilized tropical fruit canistel (Pouteria campechiana) by employing three
species of lactobacillus. The analysis of nutritive content revealed an appreciable amount of carbohydrates (36.1%) and reducing
sugars (19.8%) in canistel pulp. The highest amount of LA was produced by L. casei (20.07g/L) followed by L. acidophilus
(18.9g/L) and L. plantarum (18.9g/L).
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culture was further sub cultured in to MRS agar plates and The Lactobacillus cultures were cultivated in modified MRS
used for further studies. broth containing canistel hydrolysates instead of distilled
water. These media were kept for incubation at ambient
2.4. Preparation of hydro lysates for fermentation temperature on rotary shaker at 120 rpm for 3 days. These
Steam explosion were used as inocula for further studies.
The modified method of Pumiput et al., (2008) [7] was used
for substrate hydrolysate preparation. About 8gram of canistel 2.6. Media and fermentation conditions
pulp was steam exploded in an autoclave at 121 °C for To screen for lactic acid production from canistel
20min.Sterile water was added to the wet pretreated material hydrosylates , fermentation media containing fruit
to make the volume of 200ml and boiled at 80 °C for 30 min. hydrolysate was prepared in 250ml capacity conical flask by
Later the hydrolysate was recovered by filtration with cheese adding components of synthetic medium (Cheng 1991) [8]
cloth. (0.034g FeSO4, 1.0g Sodium acetate, 1.23g MgSO4.7H2O,
0.034g MnSO4.H2O, 0.65g KH2PO4, 30g Yeast extract) in
Acid hydrolysis 100ml of hydrolysate instead of distilled water. These flasks
Acid post hydrolysis of hydrolysate was carried out to cleave were inoculated with 5% L. plantarum, L. casei and L.
the oligosaccharides in to monomeric sugars by autoclaving at acidophillus cultures precultivated in s fruit hydrolysate based
121 °C with concentration of 1% HCl v/v for 30min media and incubated at 37 °C in a shaker incubator (120rpm)
(Pumiput, 2008) [7]. for 5 days. The product (lactic acid) formation was estimated
on daily basis.
pH adjustment
The hydrolysate from acid post hydrolysis was adjusted with 2.7. Estimation of lactic acid production
NaOH to pH 6-6.8 and the precipitate was removed by The production of lactic acid was primarily detected by
filtration with Whatmann filter paper No.1 (Pumiput, 2008) estimating the titrable acidity of the fermentation medium on
[7]
. daily basis, by titrating the fermentation medium against 1M
NaOH using phenolphthalein as indicator.
2.5. Inoculum preparation
2.8. Lactic acid downstream processing and confirmation convert complex sugars in to easily fermentable monomeric
Lactic acid present in the fermentation broth was separated (Figure 3) residues.
using the conventional Calcium lactate precipitation method
for the recovery of lactic acid. These crystals were further
used for confirmation of lactic acid using p-hydroxy diphenyl
method as described by Barnett (1951) [9]. The estimation of
L(+) and D(-) lactic acid content was done using NAD+
linked L- lactate dehydrogenase assay (using analytical kit
made by Randox labortries,UK).
3. Results
3.1. Analysis of nutritive parameters in canistel pulp
Nutritive composition of canistel pulp was tabulated (Table 1)
Fig 1: Canistel fruit
Table 1: Nutritive parameters in Canistel fruit pulp
Serial No. Nutritive Parameters Obtained Value (%)
1. Moisture Content 66.3 ± 0.21
2. Ash Content 16.5 ± 0.32
3. Protein Content 1.65 ± 0.12
4. Fiber Content 0.92 ± 0.06
5. Lipid Content 0.16 ± 0.04
7. Total Sugars 36.1 ± 0.06
8. Reducing Sugars 19.8 ± 0.14
Data represents the mean of triplicates ± standard deviation
Fig 4: Amount of Lactic Acid produced using Canistel Hydrolysate Medium (CHM)
3.4. Lactic acid downstream processing and confirmation the substrate for any fermentation process. These were in
Lactic acid produced in the fermentation broth was crystalised correlation with the results obtained by previous researchers
using conventional calcium precipitation method and analysis working with canistel fruit (Morton (1987 [10]; 1992 [11]);
of crystals through p-hydroxy diphenyl method as described Pushpa-kumara (2007) [12]. The carbohydrate and crude
by Barnett (1951) [9] revealed that irrespective of lactobacillus protein content in the canistel fruit pulp is similar to the result
strain type in all the CHM L-Lactate contributes to 20% of obtained by Attapattu et al., (2014) [13]. The acid hydrolysis
DL mixture approximately in other words the bacteria are method suggested by Pumiput et al., (2008) [7] working with
producing D: L isomers in 4:1 ratio (Table 3). lactic acid production from fruit waste was successful in
producing and extracting reducing sugars from canistel fruit
Table 3: Amount of Lactic Acid Isomers (D/L- Lactate) in CHM pulp. The gradual increase in the amount of LA produced
Lactobacillus Strain D-Lactate (g/L) L-Lactate (g/L) from first day to fourth day irrespective of the strain type is
L. plantarum 15.12 3.78 due to the gradual utilization of the reducing sugars in the
L. casei 16.63 4.15 CHM with simultaneous production of LA in the medium.
L. acidophilus 15.87 3.96 These results were in correlation with the work done by
Pushparani et al., (2012) [14] using fruit peels as substrate for
4. Discusssion LA production. The substantial decrease in the LA quantity
Evaluation of nutritive parameters in fully ripened canistel after fourth day of fermentation may be attributed to the
fruit pulp was essential to analyze its feasibility to be used as decrease in the amount of available reducing sugars left over
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for fermentation along with the reduction in the number of chicken, Tropical Agricultural Research & Extension.
bacteria due to increasing acidity of the CHM. 2014; 16(2):34-39.
14. Pushparani Mudaliyar, Latha Sharma, Chandrasekhar
5. Conclusion Kulkarni. Food waste management-lacic acid production
Many unexplored indigenous fruit species have not received by Lactobacillus species. International journal of
much attention by people inspite of its potential nutritional or advanced biological research, 2012; 2(1):34-38.
phytochemical components (Kong et al., 1980) [15]. The 15. Kong KW, Chew LY, Prasad KN, Lau CY, Amin I, Sun
present study highlights a methodology for eventual J et al. Nutritional constituents and antioxidant properties
utilization, recycling and reprocessing of an under-utilized of indigenious kembayau (Dacroydes rostrata (Blume)
fruit for the production of a commercially important organic H.J. Lam) fruits. Food Res. Int. 2010; 44:2332-2338.
acid. This could reduce the overall economics of production
and further commercialization of the process to industrial
scale can save the environment from the detrimental effects
offered by careless discharge of fruit waste.
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