Producing Bacillus from the Ripe Bananas, Jackfruit Seeds and Sweet Potato Tubers by Solid Fermentation and its Effect on the Products' Biochemical Properties
More details
Hide details
1
Department of Agriculture, Agribusiness, and Environment, Bishop Stuart University, Mbarara, Uganda
2
Department of Crop and Animal Production, Faculty of Agriculture and Environmental Sciences, Mountains of the Moon University,
Fort Portal, Uganda
Submission date: 2025-06-08
Acceptance date: 2025-06-28
Publication date: 2025-06-30
Trends in Ecological and Indoor Environmental Engineering, 2025;3(2):27-36
KEYWORDS
ABSTRACT
Background:
Fermentation is defined as a biochemical process that produces energy using microorganisms from various raw material substrates. It is an ancient form of bio-preservation of food and improvement of its nutrient content. In addition to enriching the nutritional components of plant materials and preserving food, fermentation promotes the growth of beneficial microbes that play a role in human and animal health, especially in the digestive tract, commonly known as probiotics.
Objectives:
The current study investigated the fermentation of ripe banana, jackfruit seeds and sweet potato tubers to produce Bacillus subtilis and also examined the effect of fermentation on the biochemical properties of the plant materials used in the current study. The mentioned plant materials are the most cultivated crops in many parts of Uganda and generate a lot of food waste.
Methods:
The study was carried out at the soil laboratory of Mountains of the Moon University in Fort Portal tourism western Uganda. The solid fermentation of ripe bananas, jackfruit seeds and sweet potato tubers in this study was a natural process hence no inoculation of microbes onto the sample was done. Solid fermentation involved the putting the solid substrates into fermentation vessel with limited moisture. In every 3 days, temperatures, relative humidity and pH were monitored in the fermentation vessels to establish the conditions in which microbes are growing. Gram staining was done on the sub cultured microbial colonies to view the shapes of the cells and endospore structures of the Bacillus species. There were 2 biochemical tests performed on sub cultured colonies i.e. catalase test and starch hydrolysis to confirm the presence of the presence of Bacillus in the fermented samples of ripe banana, jack fruit seeds and sweet potato tubers. For catalase tests: the microbial colonies from nutrient agar on petri-dishes of ripe banana, jackfruit seeds and sweet potato tubers cultures were transferred and smeared on microscope slides. For starch hydrolysis tests, the sub cultured microbial colonies from ripe banana, jackfruit seeds, sweet potato and mix petri-dish were transferred to starch agar.
Results:
There was decrease in carbon-nitrogen ratio due to fermentation effect except the ripe banana which experienced an increase in carbon content from 2.44 carbon: 2.8 nitrogen to 3:12 carbon: 1.7 nitrogen. In additional to that, ripe banana produced high population of microbes with colony forming units of 1.6 · 104 on day 3.4 · 104 on day 6, 1.05 · 105 and 1.08 · 105 on day 12.
Conclusion:
All the fermented samples ripe banana, jackfruit seeds, sweet potato tubers and mixed samples contained Bacillus since their microbial colonies displayed rod shaped cells and gram positive and they tested catalase. Fermentation improves the biochemical properties of ripe bananas as evidenced by increase of crude proteins and fats and carbon. For samples jackfruit seeds, fermentation negatively affect the biochemical properties since they lost carbon, crude proteins and fat.
REFERENCES (35)
1.
Adu-Kwarteng, E., Sakyi-Dawson, E. O., Ayernor, G. S., Truong, V. D., Shih, F. F., & Daigle, K. (2014). Variability of sugars in staple-type sweet potato (Ipomoea batatas) cultivars: the effects of harvest time and storage. International Journal of Food Properties, 17(2), 410–420.
https://doi.org/10.1080/109429....
2.
Angelin, J., & Kavitha, M. (2022). Molecular mechanisms behind the cold and hot adaptation in extremozymes. In Extremozymes and Their Industrial Applications (pp. 141–176). Academic Press.
https://doi.org/10.1016/B978-0....
3.
Attri, S., & Goel, G. (2023). Sterilization in bioprocesses. In Basic biotechniques for bioprocess and bioentrepreneurship (pp. 329–339). Academic Press.
https://doi.org/10.1016/B978-0....
4.
Awais, M., Shah, A. A., Hameed, A., & Hasan, F. (2007). Isolation, identification and optimization of bacitracin produced by Bacillus sp. Pakistan Journal of Botany, 39(4), 1303–1312.
5.
Badar, R. A., Carmona, J. L., Collantes, J. G., Lojo, D. R., Ocampo, S. M., Ursua, R. L., & Bercede, D. H. (2022). Staining capability of plant extracts for the identification of gram-positive and gram-negative bacteria: A systematic review. Asian Journal of Biological and Life Sciences, 11(2), 277–284.
https://doi.org/10.5530/ajbls.....
6.
Banerjee, D. K., Das, A. K., Thakur, N., Talukder, S., Das, A., Sonowal, J., ... & Sakunde, D. T. (2019). Factors affecting microbial growth in livestock products: A review. International Journal of Conservation Science, 7(3), 4017–4022.
7.
Basamma Hadimani, B. H., & Shripad Kulkarni, S. K. (2017). Ideal growth conditions for mass production of biocontrol agent Bacillus subtilis (Ehrenberg) Cohn.
8.
Gauvry, E., Mathot, A. G., Couvert, O., Leguérinel, I., & Coroller, L. (2021). Effects of temperature, pH and water activity on the growth and the sporulation abilities of Bacillus subtilis BSB1. International Journal of Food Microbiology, 337, 108915.
https://doi.org/10.1016/j.ijfo....
10.
Hossain, M. S., Rahman, W., Ali, M. S., Sultana, T., & Hossain, K. M. M. (2021). Identification and antibiogram assay of Escherichia coli isolated from chicken eggs. Journal of Bio-Science, 123–133.
https://doi.org/10.3329/jbs.v2....
11.
Kaur, P., Ghoshal, G., & Banerjee, U. C. (2019). Traditional bio-preservation in beverages: Fermented beverages. In Preservatives and preservation approaches in beverages (pp. 69–113). Academic Press.
https://doi.org/10.1016/B978-0....
12.
Laue, M., Han, H. M., Dittmann, C., & Setlow, P. (2018). Intracellular membranes of bacterial endospores are reservoirs for spore core membrane expansion during spore germination. Scientific Reports, 8(1), 11388.
https://doi.org/10.1038/s41598....
13.
Li, S., Dong, S., Fang, G., Hao, Y., & Gao, Q. (2022). Study on internal structure and digestibility of jackfruit seed starch revealed by chemical surface gelatinization. Food Hydrocolloids, 131, 107779.
https://doi.org/10.1016/j.food....
14.
Liu, D., Ge, X., Pan, D., Zheng, X., & Zhou, X. (2024). Bacillus subtilis B55 degraded the ferulic acid and p-coumaric acid and changed the soil bacterial community in soils. Journal of Applied Microbiology, 135(9), lxae243.
https://doi.org/10.1093/jambio....
15.
Mahapatra, S., Yadav, R., & Ramakrishna, W. (2022). Bacillus subtilis impact on plant growth, soil health and environment: Dr. Jekyll and Mr. Hyde. Journal of Applied Microbiology, 132(5), 3543–3562.
https://doi.org/10.1111/jam.15....
16.
Meng, L., Li, S., Liu, G., Fan, X., Qiao, Y., Zhang, A., ... & Feng, Z. (2021). The nutrient requirements of Lactobacillus acidophilus LA-5 and their application to fermented milk. Journal of Dairy Science, 104(1), 138–150.
https://doi.org/10.3168/jds.20....
17.
Mustapha, A. T., Wahia, H., Ji, Q., Fakayode, O. A., Zhang, L., & Zhou, C. (2024). Multiple‐frequency ultrasound for the inactivation of microorganisms on food: A review. Journal of Food Process Engineering, 47(4), e14587.
https://doi.org/10.1111/jfpe.1....
18.
Nayak, S. K. (2021). Multifaceted applications of probiotic Bacillus species in aquaculture with special reference to Bacillus subtilis. Reviews in Aquaculture, 13(2), 862–906.
https://doi.org/10.1111/raq.12....
19.
Nicomrat, D., & Chamutpong, S. (2016). Application of microbial community for enhancing nutritional and appealing fermented juice. Applied Mechanics and Materials, 848, 131–134.
https://doi.org/10.4028/www.sc....
20.
Olmos, J., & Paniagua-Michel, J. (2014). Bacillus subtilis a potential probiotic bacterium to formulate functional feeds for aquaculture. Journal of Microbial & Biochemical Technology, 6(7), 361–365.
http://dx.doi.org/10.4172/1948....
21.
Osdaghi, E., Taghavi, S. M., Hamidizade, M., Fazliarab, A., Hajian Maleki, H., Li, X., ... & Portier, P. (2023). Clavibacter lycopersici sp. nov.: a peach-coloured actinobacterium isolated from symptomLess tomato plant. International Journal of Systematic and Evolutionary Microbiology, 73(9), 006022.
https://doi.org/10.1099/ijsem.....
22.
Pandav, P. V., Ahire, R. S., Patil, S. R., & Pawar, S. B. (2021). Process development for high density cultivation yield for Bacillus subtilis. Culture, 6, 0–2. Available:
https://journals.ijramt.com/in....
23.
Parsons, J. B., & Rock, C. O. (2013). Bacterial lipids: metabolism and membrane homeostasis. Progress in Lipid Research, 52(3), 249–276.
https://doi.org/10.1016/j.plip....
24.
Phillips, K. M., McGinty, R. C., Couture, G., Pehrsson, P. R., McKillop, K., & Fukagawa, N. K. (2021). Dietary fiber, starch, and sugars in bananas at different stages of ripeness in the retail market. PLoS One, 16(7), e0253366.
https://doi.org/10.1371/journa....
25.
Ravindran, V., Ravindran, G., & Sivakanesan, R. (1996). Evaluation of the nutritive value of Jackseed (Artocarpus heterophyllus) meal for poultry. The Journal of Agricultural Science, 127(1), 123–130.
https://doi.org/10.1017/S00218....
27.
Sahraei, F., Ahari, H., & Kakoolaki, S. (2019). Effect of Bacillus subtilis as a probiotic on protein, lipid content, and trypsin and chymotrypsin enzymes in rainbow trout biometry (Oncorhynchus mykiss). Aquaculture International, 27(1), 141–153.
https://doi.org/10.1007/s10499....
28.
Sargantanis, J., Karim, M. N., Murphy, V. G., Ryoo, D., & Tengerdy, R. P. (1993). Effect of operating conditions on solid substrate fermentation. Biotechnology and Bioengineering, 42(2), 149–158.
https://doi.org/10.1002/bit.26....
29.
Shukla, S. K., Bernard, K. A., Harney, M., Frank, D. N., & Reed, K. D. (2003). Corynebacterium nigricans sp. nov.: proposed name for a black-pigmented Corynebacterium species recovered from the human female urogenital tract. Journal of Clinical Microbiology, 41(9), 4353–4358.
https://doi.org/10.1128/jcm.41....
30.
Soemarie, Y. B., Milanda, T., & Barliana, M. I. (2021). Fermented foods as probiotics: A review. Journal of Advanced Pharmaceutical Technology & Research, 12(4), 335–339.
https://doi.org/10.4103/japtr.....
31.
Su, Y., Liu, C., Fang, H., & Zhang, D. (2020). Bacillus subtilis: a universal cell factory for industry, agriculture, biomaterials and medicine. Microbial Cell Factories, 19(1), 173.
https://doi.org/10.1186/s12934....
32.
Timmermans, E., Bautil, A., Brijs, K., Scheirlinck, I., Van der Meulen, R., & Courtin, C. M. (2022). Sugar levels determine fermentation dynamics during yeast pastry making and its impact on dough and product characteristics. Foods, 11(10), 1388.
https://doi.org/10.3390/foods1....
33.
Wang, Q., Ren, Y., Cui, Y., Gao, B., Zhang, H., Jiang, Q., ... & Xu, C. (2022). Bacillus subtilis produces amino acids to stimulate protein synthesis in ruminal tissue explants via the phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit beta–serine/threonine kinase–mammalian target of rapamycin complex 1 pathway. Frontiers in Veterinary Science, 9, 852321.
https://doi.org/10.3389/fvets.....
34.
Yadav, S., Kaushik, R., Saxena, A. K., & Arora, D. K. (2011). Diversity and phylogeny of plant growth‐promoting bacilli from moderately acidic soil. Journal of Basic Microbiology, 51(1), 98–106.
https://doi.org/10.1002/jobm.2....
35.
Zhang, Q., Wang, Z., Gao, R., & Jiang, Y. (2025). Sugars, lipids and more: New insights into Plant Carbon sources during plant–microbe interactions. Plant, Cell & Environment, 48(2), 1656–1673.
https://doi.org/10.1111/pce.15....