Effect of Storage on Protein Composition of Fermented Soybean (Glycine Max) Seed by Bacillus Subtillis

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ISSN: 2640-9208 Research Article

Modupe Elizabeth Ojewumi*, Adebola Oyinade Odubiyi and James Abiodun Omoleye Department of Chemical Engineering, Covenant University, Nigeria

*Corresponding author: Modupe Elizabeth Ojewumi, Department of Chemical Engineering, Covenant University, Canaan land, Ogun State, Nigeria Submission: May 24, 2018; Published: August 20, 2018

Abstract The effect of storage (shelf life) on percentage protein composition of stored fermented soybean seed was studied. Samples were fermented with Bacillus substilis

the fermented samples (as a starter to monitor culture) the andeffect without of salting (natural on the fermentation) shelf life. Constant for 3 weightdays. Two was methods obtained of from preservation 12hours of were drying used, and namely; about 0% oven moisture drying contentand salting. which Each is verysample close was to oven total dried dryness within was 4-24hours obtained inand both the fermentedpercentage soybean moisture samples. content Samplesremoved were was calculated.monitored 1-3%for two salt weeks. was also The added protein to content reduced with storage duration while pH moved towards acidic medium.

Keywords: Fermentation; Glycine max; Shelf life; Physiology; Bacillus subtillis

Introduction - Soyabean (Glycine max) or soybean is a species of legume, a na- ity of the fermented product is improved. Fermented Soya has tive to East Asia and is widely grown for its edible bean which has anti-nutritional constituents [8-11]. Overall, the nutritional qual been shown to inhibit E coli innumerable uses [1]. Like most plants, soybeans grow in distinct morphological stages as they develop from seeds into fully mature infection [12], minimize fluid losses shorten the duration of diarrhoea when supplemented to the diet in ETEC-infected small intestinal segments of piglets [12-16], and of malnourished children [17,18].

plants. The first stage of growth is germination, a method which There seems to be a general agreement on the spore-forming first becomes apparent as a seed’s radical emerges [2]. Like many are extremely high in protein content. On average, dry soybean con- Bacillus legumes, soybeans can fix atmospheric nitrogen [3]. Soybean seeds During the fermentation of soya bean with systematic investigation species as the main fermentation organisms [7,19-25]. showed that Bacillus subtillis is the most dominant bacterium re- tains roughly 40% protein, 35% carbohydrate, 20% soybean oil, the highest protein content among legume species. Soy protein is a and 5% ash (non-aqueous, metal oxides). Therefore, soybean has heat-stable protein, thus allowing soy seeds to undergo high tem- species of bacillus such as Bacillus lichenioformis, Bacillus megate- sponsible for the fermentation [5,26]. Literature revealed that some perature cooking and fermentation, without destroying the entire rium, L. mesenteriodes and Staphylococcus are also found in the fer- chemical composition of the soybean. mented condiment (Iru). In a study conducted on the fermentation of Iru, it was found that Gmelinaarborea as well as banana leaves accelerated fermentation of seeds, while also bringing an increase minerals and B vitamins [4]. Soy vegetable oil, used in food and in- The beans contain significant amounts of phytic acid, dietary in protein and crude fat contents with corresponding decrease in dustrial applications, is another product of processing the soybean carbohydrate [27]. crop. Traditional non-fermented food uses of soybeans include soy milk from which tofu and tofu skin are made. Fermented soy foods Materials and Method

Source of raw materials includeFermentation soy sauce, isfermented the chemical bean breakdown paste, natto of and substance tempeh by[5,6]. bacteria, yeast or other microorganism into alcohol, carbon dioxide Soya bean seeds were bought from open market. The starter or organic acids [7]. Fermentation may also decrease or eliminate cultures used were freshly prepared in the Microbiology laboratory, Covenant University, Ota, Nigeria.

Preparation of seed for fermentation process Moisture content determination The seeds were processed according to method [28,29]{Ojew-

Method [6] was used. Sample was oven dried after fermenting The percentage moisture content was calculated until constant umi, 2017 #26;Ojewumi, 2018 #181}. for 3 days and dried between 4 to 24hrs with four hours interval. Preparation of starter culture weight was obtained from 12-24hrs with 0% moisture content. Previously isolated Bacillus subtilis was used in solid inactive - form. This was activated by taking some of the bacteria with a loop corded and stored in an air tight container. Samples were cooled in a desiccator and the final weights were re and mixing it with a freshly prepared nutrient broth and incubated Proximate analysis for 24hours. pH determination moisture content was carried out in this research. The analysis was Determination of % Protein composition using method [5] and done weekly for 2weeks (Figure 1).

ResultMethod [30,31] was used for the determination of pH.

(Figure 1-6).

Figure 1: The effect of storage duration on pH of oven dried naturally fermented Soybean.

Figure 2: The effect of sorage duration on pH of Bacillus subtilis fermented Soybean. Discussion - tional properties. The ones dried for 4- 8 hours still had moisture up to 12-16 hours showed to have better preservation of the func Percentage moisture content in them, which increased its deterioration while the ones dried for The result of this research work show that oven drying im- concentration were found to be a little low, which could indicate proves the shelf life of soybean. Effect of moisture content on its de- longer than 16 hours had no moisture in them anymore but protein that due to high temperature some of this properties of the seeds terioration was carried out and the bacteria responsible for deteri- have been lost (Figure 2). oration need moisture for their metabolism. The samples dried for

Volume - 2 Issue - 4 How to cite this article: Modupe E O, Adebola O O, James A O. Effect of Storage on Protein Composition of Fermented Soybean (Glycine Max) Seed by 196 Bacillus Subtillis. Nov Tech Nutri Food Sci. 2018: 2(4). NTNF.000543. DOI: 10.31031/NTNF.2018.02.000543 Nov Tech Nutri Food Sci Copyright © Modupe Elizabeth Ojewumi

Figure 3: Protein analysis for Bacillus subtilis fermented oven dried samples.

Figure 4: Protein analysis for naturally fermented oven dried samples.

Figure 5: Protein analysis for Bacillus subtilis fermented and salted samples.

Volume - 2 Issue - 4 Modupe E O, Adebola O O, James A O. Effect of Storage on Protein Composition of Fermented Soybean (Glycine Max) Seed by How to cite this article: 197 Bacillus Subtillis. Nov Tech Nutri Food Sci. 2018: 2(4). NTNF.000543. DOI: 10.31031/NTNF.2018.02.000543 Nov Tech Nutri Food Sci Copyright © Modupe Elizabeth Ojewumi pH analysis

Table 1: Percentage moisture content removed for naturally fermented oven dried samples.

Time (Hrs) Initial Weight (G) Final Weight (G) % Moisture Content Removed 4 20 8.9

8 20 34.522 12 20 6.42.4 2 20 2 0

1620 20 2 0 24 20 2 0

Table 2: Percentage moisture content removed for Bacillus Subtilis fermented oven dried samples.

Time (hrs) Initial Weight (g) Final Weight (g) % Moisture Content Removed 4 28

8 35 65 12 35 16 30.71 35 5.27 0.05 1620 35 5.27 0.050 24 35 5.25 0 Table 1 & 2 above shows the effect of storage35 duration on the three, and salt5.25 was added as a preservative to each of the sample pH of both fermented samples. The higher the pH the lower the

in the ratio 1%: 2%: 3%, Figure 5 & 6. It was observed that after strive in an acidic medium. Increase in pH towards the acidic range the highest protein concentration, by the second week, the three risk and rate of spoilage, since microorganisms finds it difficult to the first week of storage, sample with 3% salt concentration had was recorded for drying durations 8-24 hours while increase in samples all had very close concentration, the protein concentration pH towards alkaline medium was noticed at 4 hour. The increase - in pH observed in this study is an indication that fermentation tration had increased. From the result the sample preserved by 2% of the 3% sample had reduced while the sample with 1% concen still continued after the processing period of soybean to a protein salt had a higher protein concentration and maintained all through the storage period. In the control sample colour change and maggot organisms responsible for the fermentation are still present at con- infestation was noticed on the third day of storage. condiment (post fermentation operation). This confirmed that the Conclusion sumptionProtein Analysis [6]. This work concluded that to increase the shelf life of fermented Oven dried samples fermented using Bacillus subtillis was mon- African locust bean seeds, water must be totally eliminated from it. itored for two weeks and protein analysis were carried out every Total dryness of the fermented condiment will preserve the seed week on each sample. The protein concentration was determined - by tracing the absorbance on the standard absorbance-concen- mulation. This will reduce scarcity (due to very short shelf life) and and increase its utilization especially in protein deficient food for tration protein graph and the values were multiplied by 10 using improve its availability all year round. Drying will also get rid of the undesirable smell which has made Ire unpopular in the urban four hours showed a higher protein concentration but as storage region. the conversion formula, Figure 3. From week 1 sample dried for time increased the concentration of the protein began to vary, it The application of many advanced food preservation methods increased and then decreased for each sample all through the stor- such as canning, refrigeration and irradiation have greatly reduced because of the high cost in their operations. Moisture content re- concentration with storage although the concentration varied too age period. Sample dried for 16 hours had a more stable protein with storage time compared to the samples dried at 4,8 12,20 and 24 hours. Referencesduction was found to be an efficient method for preservation. Also on naturally fermented oven dried samples Figure 4, the 1. results shows that protein concentration for each of the dried sam- línea]. University of Melbourne. Institute for Land & Food Resources. Melbourne,Porcher M (2005)Australia. Multilingual multiscript plant name database [en ples were higher compared to the bacillus fermented oven dried samples. From the graph the protein concentration varied with 2. Purcell LC, Salmeron M, Ashlock L (2014) Soybean growth and development. Arkansas Soybean Handbook. University of Arkansas, Fayetteville, USA. concentration compared to the samples dried for 4, 8, 12, 20 and 24 storage time. Samples dried for 16 hours had the higher protein hours. Samples with and without starter culture was divided into 3. Canfield DE, Glazer AN, Falkowski PG (2010) The evolution and future of earth’s nitrogen cycle. Science 330(6001): 192-196. Volume - 2 Issue - 4 How to cite this article: Modupe E O, Adebola O O, James A O. Effect of Storage on Protein Composition of Fermented Soybean (Glycine Max) Seed by 198 Bacillus Subtillis. Nov Tech Nutri Food Sci. 2018: 2(4). NTNF.000543. DOI: 10.31031/NTNF.2018.02.000543 Nov Tech Nutri Food Sci Copyright © Modupe Elizabeth Ojewumi

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Volume - 2 Issue - 4 Modupe E O, Adebola O O, James A O. Effect of Storage on Protein Composition of Fermented Soybean (Glycine Max) Seed by How to cite this article: 199 Bacillus Subtillis. Nov Tech Nutri Food Sci. 2018: 2(4). NTNF.000543. DOI: 10.31031/NTNF.2018.02.000543