Molecular screening of potential Bacillus species bacteriocin starter cultures isolated from Nigerian fermented
), Ilesanmi F. Fadahunsi(2), Femi Ayoade(3), Janet O. Olaitan(4),
(1) Osun State University, Osogbo & University of Ibadan
(2) University of Ibadan
(3) Redeemers University, Ede & African Centre of Excellence for the Genomics of Infectious Diseases (ACEGID
(4) Osun State University, Osogbo
Corresponding Author
Abstract
Traditional fermented food condiments in Nigeria are majorly produced by using protein-rich leguminous seeds via spontaneous fermentation or back slopping. However, health risks such as the production of biogenic amines, presence of toxigenic genes and food borne pathogens remains major concerns of public health significance. Hence, the need for the production of safe condiments using non-pathogenic food-grade starter cultures. Isolation and characterization of organisms from spontaneously fermented condiments made from Parkia biglobosa, Ricinus communis, Pentaclethra macrophylla and Prosopis africana was done using standard methods. Phenotypic and biochemical characterizations was down using established protocols while molecular identification was done using 16S rRNA sequencing. Results obtained showed that the microbial load (cfu/ml) recorded from the spontaneously fermented condiments ranged from: 2.1×107-5.2×1012, 15.2×107-4.4×1010, 4.4×1010-4.4×1012 and 6.4×107-1.6×1012 for P. biglobosa, R. communis, P. macrophylla and P. africana respectively. Identification of isolates revealed Bacillus. However, 22.8% of the Bacillus isolates harboured HblA and HblD genes while 40.9 % showed bands for the presence of HblC genes. In addition, the percentage occurrence of 18.1%, 27.2% and 50% was obtained for the presence of NheA, NheB and NheC genes respectively while none of the Bacillus isolates showed bands for the presence of Cytk and EM1 genes. More so, five Bacillus isolates were identified to be non-toxigenic, showing bands only for Bac, Osbo and Spas genes which are bacteriocin producing genes. Put together, the identified non-toxigenic food-grade Bacillus species recorded in this study can be recommended as potential bacteriocin starter cultures for the fermentation of microbiologically safe food products.
Keywords
References
Adedeji BS, Ezeokoli OT, Ezekiel CN, Obadina AO, Somorin YM, Sulyok M, Adeleke RA, Warth B, Nwangburuka CC, Omemu AM, Oyewole O, & Krska R (2017). Bacterial species and mycotoxin contamination associated with locust bean, melon and their fermented products in south-western Nigeria. International Journal of Food Microbiology. 258 (2017):73–80.
Adekoya IA, Obadina M, Olorunfemi O, Akande S, Landschoot S, De Saeger S, & Njobeh P (2019). Occurrence of bacteria and endotoxins in fermented foods and beverages from Nigeria and South Africa. International Journal of Food Microbiology 305 (2019):108251.
Ademola OM, Adeyemi TE, Ezeokoli OT, Ayeni KI, Obadina AO, Somorin YM, Omemu AM, Adeleke RA, Nwangburuka C, Oluwafemi CF, Oyewole O, & Ezekiel CN (2018). Phylogenetic analyses of bacteria associated with the processing of iru and ogiri condiments. Letters in Applied Microbiology. 67 (4):354–62.
Ahaotu I, Anyogu A, Njoku OH, Odu NN, Sutherland JP, & Ouoba LII (2013). Molecular identification and safety of Bacillus species involved in the fermentation of African Oil beans (Pentaclethra macrophylla Benth) for production of ugba. International Journal of Microbiology. 162 (2013): 95-104.
Almasoud N, Xu Y, Nicolaou N, & Goodacre R (2014). Optimization of matrix assisted desporption/ionization time of flight mass spectrometry (MALDI-TOF-MS) for the characterization of Bacillus and Brevibacillus species. Analytica Chemica Acta. 840: 49-57. https://doi.org/10.https://doi.org/10.1016/j.aca.2014.06.032. PMID: PMC4223412.
Amoa-Awua WK, Awusi B, Owusu M, Appiah V, Ofori H, Thorsen L, & Jespersen L (2014). Reducing the atypical odour of dawadawa: Effect of modification of fermentation conditions and post-fermentation treatment on the development of the atypical odour of dawadawa. Food Control. 42 (2014):335–42.
Batiha GE, Hussein DE, Algammal AM, George TT, Jeandet P, Al-Snafi AE, Tiwari A, Pagnossa JP, Lima CM, Thorati ND, Zhoor MM El-Esawi M, Dey A, Alghamdi S, Hetta HF, & Cruz-Martins N (2021). Application of natural antimicrobials in food preservation: Recent reviews. Food Control.126 (2021): 1-14. https://doi.org/10.1016/j.foodcont.2021.108066.
Butkhot N, Soodsawaeng P, Vuthiphanchai V, & Nimrat S (2019). Characterization and biosafety evaluation of a novel bacteriocin produced by Bacillus velezensis BUU004. International Food Research Journal. 26(5): 1617-1625.
Caulier S, Nannan C, Gillis A, Licciardi G, Bragard C, & Mahillon J (2019). Overview of the antimicrobial compounds produced by members of the Bacillus subtilis group. Frontiers in Microbiology. 10(4): 1-20. Doi: 10.3389/fmicb.2019.00302.
Celandroni A, Vecchione A, Cara A, Mazzantini D, Lupetti A, & Ghelardi E (2019). Identification of Bacillus species: implication on the quality of probiotic formulation. PlusOne. 14(5):1-13. https://doi.org/10.1371/journal.pone.0217021.
Compaore CS, Jensen LB, Diawara B, Quedraogo GA, Jakobsen M, & Ouoba LII (2013). Resistance to antimicrobials and acid bile tolerance of Bacillussp isolated from Bikalga, fermented seeds of Hibiscus sabdariffa. African Journal of Food science. 7(2013): 408-413.
da Silva SS, Lopes AM, de Carvalho SEV, de Oliveira RYC, & de Souza ORP (2018). Bacteriocin partitioning from a clarified fermentation broth of Lactobacillus plantarum ST16Pa in aqueous two-phase systems with sodium sulfate and choline-based salts as additives. Process Biochemistry. 66(1): 212–221.
Dabire Y, Mogmenga I, Somda MK, Ugwuanyi J, Ezeogu LI, Dicko MH, & Traore AS (2020). Production technique, safety and quality of soumbala, a fermented food condiment sod and consumed in Burkina-Faso. African Journal of Science. 14(2): 38-52. Doi: 10.5897/AJFS2019.1891.
Dabire Y, Somda MK, Ugwuanyi J, Ezeogu LI, & Traore AS (2018). Diversity of Bacillus species and their antimicrobial compounds involved in alkaline fermentation of indigenous food condiments used in Africa. International Journal of Advanced Research.6(12):2320-5407. DOI:10.21474/IJAR01/8147.
Dabire Y, Somda N, Compaore CS, Mogmega I, Somda MK, Ouattara AS, Dicko MH, Ugwuanyi JO, Ezeogu LI, & Traore AS (2021). Molecular screening of bacteriocin-producing Bacillus spp. isolated from Soumbala, a fermented food condiment from Parkia biglobosa seeds. Scientific African. 12(2021): e00836.
Delesa DA (2017). Bacteriocin as an advanced technology in food industry. International Journal of Advanced Research in Biological Sciences. 12(4): 178-190. https://doi.org/10.22192/ijarbs.2017.04.12.018
Dimidi E, Cox SR, Rossi M, & Whelan K (2019). Fermented foods: definitions and characteristics, impact on the gut microbiota and effects on gastrointestinal health and disease. Nutrients 11(8):1806.
Epparti P, Eligar SM, Sattur AP, Gnanesh-Kumar BS, & Halami PM (2022). Characterization of dual bacteriocin producing Bacillus subtilis SC3.7 isolated from fermented food. LWT-Food Science and Technology.154(2022): 112854.
Fernandez B, LeLay C, & Jean J (2013). Growth, acid production and bacteriocin production by probiotic candidates under stimulated colonic conditions. Journal of Applied Microbiology. 114(2): 877-85.
Fira D, Dimkic I, Beric T, Lozo J, & Stankovic S (2018). Biological control of plant pathogens by Bacillus sp. Journal of Biotechnology. 10(285): 44-55.
Flynn K, Villarreal BP, Barranco A, Belc N, Bjornsdottir B, Fusco V, Rainieri S, Smaradottir SE, Smue I, Teixeira P, & Jorundsdottir HO (2019). An introduction to current food safety needs. Trends in Food Science and Technology. 84(3): 1-3. https://doi.org/10.1016/j.ijfoodmicro.2007.06.001.
Gopikrishan T, Keerthana H, Kumar S, Perumal K, & Elangovan E (2021). Impact of Bacillus in fermented soybean foods on human health. Annals of Microbiology. 71(2021): 30-46.
Kang SJ, Park SJ, & Mishig-Ochir T (2014). Antimicrobial peptides: Therapeutic potentials. Expert Review of Anti-infective Therapy. 12(1): 1477-1486.
Kaskoniene V, Stankevicius M, Bimbiraite-Surviliene K, Naujokaityte G, Serniene L, Kondrotiene K, Malakauskas M, & Maruska A (2017). Current state of purification, isolation and analysis of bacteriocins produced by lactic acid bacteria. Applied Microbiology and Biotechnology. 101(4): 1323–1335.
Kim JH, Lee ES, Kim BM, Ham JS, & Oh MH (2023). Simple purification and antimicrobial properties of bacteriocin-like inhibitory substance from Bacillus species for the biopreservation of cheese. Foods. 13(2023): 10-24.
Kimura K & Yokoyama S (2019). Trends in the application of Bacillus in fermented foods. Current Opinion in Biotechnology. 56C: 36-42.
Laranjo M, Potes ME, & Elias M (2019). Roles of starter cultures on the safety of fermented meat products. Frontiers in Microbiology. 10(853): 1-11. 10.3389/fmicb.2019.00853.
Maina J, Matharan JM, Kikuvi GM, & Wafula E (2021). Isolation and identification of bacteriocin producing Bacillus sp from Rastrineobola argentea (Omena) with activity against Bovine mastitis bacterial pathogens. Journal of Food Security. 9(2): 62-75.
Meade E, Slattery MA, & Garvey M (2020). Bacteriocins, potent antimicrobial peptides and the fight against multi drug resistant species: Resistance is futile? Antibiotics: 9(1): 32.
Mishra SS, Ray RC, Panda SK, & Montet D (2017). Technological innovations in processing of fermented foods, an overview (chapter 1). In: Ray, R.C. and Montet, D. (Eds). Fermented foods, part II. Technological interventions. Food Biology series. Taylor and Francis Group. 21-45.
Mohammed Y, Lee B, Kang Z, & Du G (2014). Development of a two-step cultivation strategy for the production of vitamin B12 by Bacillus megaterium. Microbial Cell Factory. 13(1): 102-110. https://doi.org/10.1186/s12934014-0102-7. PMID: 25023574.
Nath S, Chowdhury S, & Dora KC (2015). Application of Bacillus species as a biopreservative for food preservation. International Journal of Engineering Research and Application. 5(4): 85-95.
Nwagu TN, Ugwodo CJ, Onwosi CO, Inyima O, Uchendu OC, & Akpuru C (2020). Evaluation of the probiotic attributes of Bacillus strains isolated from traditional fermented African locust bean seeds (Parkia biglobosa), daddawa. Annals of Microbiology. 70(20): 30-39.
O’Connor PM, Ross RP, Hill C, & Cotter PD (2015). Antimicrobial antagonists against food pathogens: a bacteriocin perspective. Current Opinions in Food Science. 2(5): 51-57.
Obadina AO, Oyewole OB, Sanni LO, Tomlins KI, & Westby A (2008). Identification of hazards and critical control points (CCP) of cassava fufu processing in South-West Nigeria. Food Control. 19(1): 22-6.
Ogwuanyi JO & Okpara A (2019). Current status of alkaline fermented foods and seasoning agents in Africa. New Advances on Fermentation Processes. Doi: 10. 5772.intechopen87052. 1-34.
Olasupo NA & Okorie PC (2019). African fermented food condiments: microbiology impacts on their nutritional values. Frontiers and New Trends in the Science of Fermented Foods and Beverages. Doi: 10.5772intechopen.83466. 21-42.
Ouoba LII, Thorsen L, & Varnam AH (2008). Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from soumbala and bikalga, African alkaline fermented food condiments. International Journal of Food Microbiology. 124(2008): 224-230.
Owusu-Kwarteng J, Parkouda C, Adewumi GA, Ouoba LII, & Jespersen L (2020). Technologically relevant Bacillus species and microbial safety of West African traditional alkaline fermented seed condiments. Critical Reviews in Food Science and Technology. 62(3): 1-18.
Pershakova TV, Kupin GA, Kabalina DV, Mikhaylyuta LV, Gorlov SM, & Babakina A (2018). Studying the antagonistic properties of B. subtilis bacteria to pathogens of fruits in in-vitro and in-vivo experiments. Journal of Pharmaceutical Sciences and Research. 10(4): 920-925.
Rai M, Pandit R, Gaikwad S, & Kövics G (2016). Antimicrobial peptides as natural bio-preservative to enhance the shelf-life of food. Journal of Food Science and Technology. 53(2): 3381–3394.
Sabate DC & Audisio MC (2013). Inhibitory activity of surfactin produced by different Bacillus subtilis subsp. subtilis strains against L.monocytoegenes sensitive and bacteriocin-resistant strains. Microbiology Research. 168(3): 125-129.
Sharmila PS & Vidya AK (2015). Characterization and antbacterial activity of bacteriocin producing Bacillus subtilis isolated from raw milk. International Journal on Applied Bioengineering. 9(2): 1-6. https://doi.org/10.15613/SIJRS/2015/V212/114284.
Shu LJ & Yang YL (2017). Bacillus classification based on matrix-assisted laser desporption ionization time-of-flight mass spectrometry-effects of culture conditions. Scientific Reports. 7(1): 15546-9. https://doi.org/10.1038/s41598-017-15808-5. PMID: 29138467.
Takano H (2016). The regulatory mechanism underlying light-inducible production of carotenoids in non-phototrophic bacteria. Bioscience, Biotechnology and Biochemistry. 80(2): 1264-73. https://doi.org/10.1080/09168451. PMID: 26967471.
Tanaka K, Takanaka S, & Yoshida K (2014). A second generationBacillus cell factory for rare inositol production. Bioengineered. 5(5): 331-4. https://doi.org/10.4161/bioe.29897. PMID: 25482235.
Thorsen L, Azokpota P, Hansen BM, Ronsbo MH, Nielsen KF, Hounhouigan DJ, & Jakbosen M (2011). Formation of cereulide and enterotoxins by B. cereus in fermented African locust beans. Food Microbiology. 28(8): 1441-7.
Villeger R, Cachon R & Urdaci MC (2017). Fermented foods, microbiology, biochemistry and biotechnology. In: Ray, R.C., Montet, D (Eds). Fermented Foods, part II. Technological interventions. Food Biology Series. Boca Raton, Taylor and Francis Group. 1-20.
Wang J, Xu H, Liu S, Song B, Liu H, Li F, Deng S, Wang G, Zeng H, Zeng X, Xu D, Zhang B, & Xin B (2021). Toyocin, a novel leaderless bacteriocin that is produced by Bacillus toyonensis Xin-YC13 and specially targets B. cereus and L.monocytogenes. Applied Environmental Microbiology. 87(12): 1-12.
Youcef-Ali M, Chaouche NK, Dehimat L, Bataiche I, Ali MK, Cawoy H, & Thonart P (2014). Antifungal activity and bioactive compounds produced by Bacillus mojavensis and B. subtilis. African Journal of Microbiology Research. 8(6): 476-484.
Article Metrics
Abstract View
: 44 times
Download : 9 times
DOI: 10.57046/LEGP3275
Refbacks
- There are currently no refbacks.
Copyright (c) 2024 Proceedings of the Nigerian Academy of Science
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
