| Peer-Reviewed

Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates

Received: 5 January 2022    Accepted: 24 January 2022    Published: 9 February 2022
Views:       Downloads:
Abstract

Amylases are known to be of great importance worldwide. Most of their production using refined carbon and nitrogen substrates are expensive, resulting in the production of expensive goods which are not easily affordable by end users in our country Nigeria. This necessitates the call for the use of cost-effective substrates under optimized condition for maximum amylase yield. Hence, the present study was focused on the optimization of amylase production from Lactobacillus plantarum CS using agro wastes from local substrates. The parameters assayed were carbon and nitrogen substrates from agro wastes, fermentation period (24-168h), pH (3.5-11) and temperature (30°C-70°C). Others included solvents for preparation of fermentation medium and metal ions of Mg2+, Mn2+, K+, Na+, Hg2+, Fe2+, Cu2+ and Pb2+ as supplements. The amylase produced via shake-flask fermentation was assayed using Dinitro salicylic acid method. The result revealed 3% sweet potato and 1% bambara nut wastes as the best carbon and nitrogen components of the required fermentation medium of pH 6.5. The optimal amylase produced (38.6U/ml) was achieved within 48h at 35°C with 1% inoculum and Mn2+/ Mg2+ as a co-factor. Relatively, Cu2+, Pb2+ and Hg2+ reduced the amylase yield to < 68.13% as against Mn2+ and Mg2+ which enhanced the production to 121.66% and 101.09% respectively. Natural rain water used as solvent for the preparation of fermentation medium significantly enhanced amylase production (24.65 U/ml) in relation to other test waters at P>0.05. Comparatively, there was a significant increase in amylase produced under optimized condition than not. Conclusively, the obtained results which revealed high amylase yield with available and cost effective agro-wastes is promising for industrial application in Nigeria.

Published in Advances in Bioscience and Bioengineering (Volume 10, Issue 1)
DOI 10.11648/j.abb.20221001.12
Page(s) 11-18
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Amylase, L. plantarum CS, Optimization, Agro Wastes, Fermentation

References
[1] Elmansy, E. A., Asker, S. M., El-kady, E. M., Hassanein S. M., & El-Beih, F. M. (2018). Production and optimization of -amylase from thermo-halpohilic bacteria isolated from different local marine environment. Bulletin of the National Research Centre, 42: 31. https://doi.org/10.1186/s42269-018-0033-2
[2] Abou-Elela, G., El-Sersy, N., & Wefky, S. (2009). Statistical optimization of cold adapted alpha-amylase production by free and immobilized cells of Nocardiopsis aegyptia. Journal of Applied Science Research, 5 (3): 286–292. https://www.researchgate.net/publication/286716117_Statistical_optimization_of_cold_adapted_a-amylase_production_by_free_and_immobilized_cells_of_Nocardiopsis_aegyptia
[3] Gupta, R., Gigras, P., Mohapatra, H., Goswami, V., & Chauhan, B. (2003). Microbial α-amylases: A biotechnological perspective. Process Biochemistry, 38: 1599–1616. Doi 10.1016/S0032-9592 (03)00053-0.
[4] Gouda, M., & Elbahloul, Y. (2008). Statistical optimization and partial characterization of amylase production. World Journal of Agricultural Science, 4: 359–368. https://www.idosi.org/wjas/wjas4 (3)/13.pdf
[5] Ahmadia, A., Ghobadia, S., Khajehb, K., Nomanpourc, B., & Dalfard, B. (2010). Purification of α-amylase from Bacillus sp. GHA1 and its partial characterization. Journal of the Iranian Chemical Society, 7: 432–440. https://doi.org/10.1007/BF03246029
[6] Viader, R., Yde, M., Hartvig, J. Pagenstecher, M., Carlsen, J., Christensen, T., & Andersen, M. (2021). Optimization of beer brewing by monitoring α-amylase and β-amylase activities during mashing. Beverages, 7 (1): 13. https://doi.org/10.3390/beverages7010013
[7] Sajid, Z., Shafique, M., Naz, S., & Jabeen, N. (2020). Microbial amylases: Production from nonconventional sources of starch and multifarious applications. Pakistan Journal of Biochemistry and Molecular Biology, 53 (4), 70-85. http: //www.pjbmb.com/index.php/pjbmb/article/view/5
[8] Pandey, A, Nigam, P., Soccol, C., Soccol, V., Singh, D., & Mohan, R. (2000). Advances in microbial amylases. Biotechnology Applied Biochemistry, 31: 135-152. DOI: 10.1042/ba19990073.
[9] Nithya, K., Muthukumar, C., Kadaikunnan, S., Alharbi, N., Khaled, J., & Dhanasekaran, D. (2017). Purification, characterization, and statistical optimization of a thermostable alpha-amylase from desert actinobacterium Streptomyces fragilis DA7-7. 3 Biotechnology, 7 (5): 350. doi: 10.1007/s13205-017-0981-5.
[10] Sundarram, A., & Murthy, T. (2014). Alpha-amylase production and applications: A review. Journal of Applied Environmental Microbiology, 2 (4): 166–75. doi: 10.12691/jaem-2-4-10.
[11] Karim, K., Husaini, A., Sing, N., Sinang, F., Roslan, H., & Hussain, H. (2018). Purification of an alpha amylase from Aspergillus flavus NSH9 and molecular characterization of its nucleotide gene sequence. 3 Biotechnology, 8 (4): 204. doi: 10.1007/s13205-018-1225-z.
[12] Sachdev, S., Ojha, S., & Mishra, S. (2016). Bacillus Spp. amylase: Production, isolation, characterisation and its application. International Journal of Applied Sciences and Biotechnology, 4 (1), 3–14. https://doi.org/10.3126/ijasbt.v4i1.14574
[13] Sanni, A., Morlon-Guyot, J., & Guyot, J. (2002). New efficient amylase-producing strains of Lactobacillus plantarum and L. fermentum isolated from different Nigerian traditional fermented foods. International Journal of Food Microbiology, 72 (1-2): 53-62. doi: 10.1016/s0168-1605(01)00607-9.
[14] Khusro, A., Barathikannan, K., Aarti, C., & Agastian, P. (2017). Optimization of Thermo-Alkali Stable Amylase Production and Biomass Yield from Bacillus sp. under Submerged Cultivation. Journal of Fermentation, 3 (1): 7. doi: 10.3390/fermentation3010007.
[15] Saha, K., Maity, S., Roy, S., Pahan, K., Pathak, R., Majumdar, S., & Gupta, S. (2014). "Optimization of Amylase Production from B. amyloliquefaciens (MTCC 1270) Using Solid State Fermentation", International Journal of Microbiology, vol. 2014, Article ID 764046, 7 pages, 2014. https://doi.org/10.1155/2014/764046
[16] George – Okafor, U., Nwachukwu, U., & Mike – Anosike, E. (2018). Screening and identification of predominant Lactobacillus spp from fermented milled-mixed corn – soybean wastes. International Journal of Scientific and Engineering Research, 9 (2): 2050 – 2060. https://www.ijser.org/researchpaper/Screening-and-Identification-of-Predominant-Lactobacillus-spp-from-Fermented-Milled-Mixed-Corn-Soybean-Wastes.pdf
[17] Bogale, A. T., & Prapulla, S. G. (2015). Studies on amylase from probiotic lactic acid bacteria. International Journal of Modern Chemistry & Applied Science, 2 (4): 211-214. https://aip.scitation.org/doi/pdf/10.1063/1.5050161
[18] Ekka, A., & Namdeo, N. (2018): Screening, isolation and characterization of amylase producing bacteria and optimization for production of amylase. Journal of Biotechnology and Biochemistry, 4 (2): 50 – 56. DOI: 10.9790/264X-0402015056.
[19] Simair, A., Khushk, I. Qureshi, A., Bhutto, M., Chaudhry, H., Ansari, K., & Lu, C. (2017). Amylase production from thermophilic Bacillus sp. BCC 021-50 isolated from a marine environment. Fermentation, 3 (2): 25. https://doi.org/10.3390/fermentation3020025
[20] Singh, R., Kapoor, V., & Kumar, V. (2012). Utilization of Agro-industrial wastes for the simultaneous production of amylase and xylanase by thermophilic actinomycetes. Brazilian Journal of Microbiology, 43 (4): 1545-1552. doi: 10.1590/S1517-838220120004000039.
[21] Singh, S., Tyagi, C., Dutt, D., & Upadhyaya, J. (2009). Production of high level of cellulase-poor xylanases by wild strains of white-rot fungus Coprinellus disseminates in solid-state fermentation. New Biotechnology, 26 (3-4): 165-170. doi: 10.1016/j.nbt.2009.09.004.
[22] Mushtaq, Q., Irfan, M., Tabssum, F., & Qazi, I. (2016). Potato peels: A potential food waste for amylase production. Journal of Food Process Engineering 40 (4): 1-8. DOI: 10.1111/jfpe.12512.
[23] Sanchez, S., & Demain, A. (2008). Metabolic regulation and overproduction of primary metabolites. Microbial Biotechnology, 1 (4): 283-319. Doi: 101111/j.1751-7915.2007.00015.x.
[24] Vidyalakshmi, R., Paranthaman R., & Indhumathi, J. (2009). Amylase production on submerged fermentation by Bacillus spp. World Journal of Chemistry 4 (1): 89 – 91. https://www.idosi.org/wjc/4 (1)09/14.pd
[25] Panda, S., Swain, M., Kar, S., Ray, R., & Montet, D. (2008). Statistical optimization of α-amylase production by probiotic Lactobacillus plantarum MTCC 1407 in submerged fermentation. Polish Journal of Microbiology, 57 (2): 149-155. https://www.researchgate.net/publication/51420227_Statistical_optimization_of_a-amylase_production_by_probiotic_Lactobacillus_plantarum_MTCC_1407_in_submerged_fermentation
[26] Gupta, R. Hiteshi, K., & Didwal, G. (2018). Production, optimization of –amylase from Bacillus licheniformis. Journal of Advaned Research in Biology & Pharmacy Research, 1 (5): 1 – 15.·https://www.researchgate.net/publication/327237754_Production_optimization_of_a-amylase_from_Bacillus_licheniformis
[27] Nwagu, T. N., & Okolo, B. N. (2011). Extracellular amylase production of a thermotolerant Fusarium spp. isolated from eastern Nigeria soil. Brazillian Archives of Biology and Technology 54 (4): 649-658. DOI: 10.1590/S1516-89132011000400002.
[28] Nyarko, C., Mills, J., Afortude, J., Kizzie, N., Agbale, C. M., & Nyarko, S. B. (2019). Effect of pH stability on alpha amylase extracted from Aspergillus niger on starch from local rice in Ghana. Asian Journal of Microbiology and Biotechnology, 4 (1), 24-34.
[29] Giraud, E., Brauman, A., Keleke, S., Lelong, B., & Raimbault, M. (1991). Isolation and physiological study of an amylolytic strain of Lactobacillus plantarum. Applied Microbiology and Biotechnology, 36 (3), 379-383.
[30] Ruby, U., Serrano, A., & Traifalgar, R. (2012). Effect of pH on amylase, cellulase and protease of the Angelwing clam, Pholas orientalis. European Journal of Experimental Biology, 2 (6): 2280-2285.
[31] Hassan, M., Marzan, L., Hosna A., Hakin, A., & Azad, A. (2017). Optimization of some fermentation condition for the production of extracellular amylases by using Chryseobacterium and Bacillus isolated from organic kitchen waste. Journal of Genetic Engineering and Biotechnology, 15: 59 – 68. doi: 10.1016/j.jgeb.2017.02.009.
[32] Divakaran, D., Chandra, A., & Chandran R., (2011). Comparative study on production of – amylase from Bacillus licheniform strain. Brazilian Journal of Microbiology, 42 (4): 1397 – 1404. DOI: 10.1590/S1517-838220110004000022.
[33] Li, W., Zhou, X. and Lu, P. (2005). Structural features of thermozymes. Biotechnology Advance, 23: 271 – 281. DOI: 10.1016/j.biotechadv.2005.01.002.
[34] Shanthi, V., & Roymon, M. (2015). Response surface optimization of xylanase production by indigenous thermoalkalophillic Bacillus sp. African Journal of Microbiology Research, 9 (27): 1699-1705. https://doi.org/10.5897/AJMR2015.7557
[35] Sirohi, R., & Prakash V., (2015). Effect of metal ions on amylase production using Bacillus subtilis isolated from soil of Almora district, Uttarakhand, India. International Journal of Pure and Applied Bioscience 3 (4): 37-41.
[36] Linden, A., Mayans, O., Meyer-Klaucke, W., Antranikian, G., & Wilmanns, M. (2003). Differential regulation of a hyperthermophilic α-amylase with a novel (Ca, Zn) two-metal center by zinc. Journal of Biological Chemistry, 278 (11), 9875-9884.
[37] Arekemase, M. O., Agbabiak, T. O., Ajide-Bamigboye, N. T., Aderoboye, O. Y., Orogu, J. O., & Ahmed, T. (2020). Optimization of amylase produced from bacteria isolated from cassava peel dumpsite using submerged fermentation. Science World Journal, 15 (1): 64-75.
Cite This Article
  • APA Style

    George-Okafor Uzoamaka Ogechi, Nwachukwu Ujunwa Felicia, Ezeme-Nwafor Amara. (2022). Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates. Advances in Bioscience and Bioengineering, 10(1), 11-18. https://doi.org/10.11648/j.abb.20221001.12

    Copy | Download

    ACS Style

    George-Okafor Uzoamaka Ogechi; Nwachukwu Ujunwa Felicia; Ezeme-Nwafor Amara. Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates. Adv. BioSci. Bioeng. 2022, 10(1), 11-18. doi: 10.11648/j.abb.20221001.12

    Copy | Download

    AMA Style

    George-Okafor Uzoamaka Ogechi, Nwachukwu Ujunwa Felicia, Ezeme-Nwafor Amara. Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates. Adv BioSci Bioeng. 2022;10(1):11-18. doi: 10.11648/j.abb.20221001.12

    Copy | Download

  • @article{10.11648/j.abb.20221001.12,
      author = {George-Okafor Uzoamaka Ogechi and Nwachukwu Ujunwa Felicia and Ezeme-Nwafor Amara},
      title = {Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates},
      journal = {Advances in Bioscience and Bioengineering},
      volume = {10},
      number = {1},
      pages = {11-18},
      doi = {10.11648/j.abb.20221001.12},
      url = {https://doi.org/10.11648/j.abb.20221001.12},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.abb.20221001.12},
      abstract = {Amylases are known to be of great importance worldwide. Most of their production using refined carbon and nitrogen substrates are expensive, resulting in the production of expensive goods which are not easily affordable by end users in our country Nigeria. This necessitates the call for the use of cost-effective substrates under optimized condition for maximum amylase yield. Hence, the present study was focused on the optimization of amylase production from Lactobacillus plantarum CS using agro wastes from local substrates. The parameters assayed were carbon and nitrogen substrates from agro wastes, fermentation period (24-168h), pH (3.5-11) and temperature (30°C-70°C). Others included solvents for preparation of fermentation medium and metal ions of Mg2+, Mn2+, K+, Na+, Hg2+, Fe2+, Cu2+ and Pb2+ as supplements. The amylase produced via shake-flask fermentation was assayed using Dinitro salicylic acid method. The result revealed 3% sweet potato and 1% bambara nut wastes as the best carbon and nitrogen components of the required fermentation medium of pH 6.5. The optimal amylase produced (38.6U/ml) was achieved within 48h at 35°C with 1% inoculum and Mn2+/ Mg2+ as a co-factor. Relatively, Cu2+, Pb2+ and Hg2+ reduced the amylase yield to 2+ and Mg2+ which enhanced the production to 121.66% and 101.09% respectively. Natural rain water used as solvent for the preparation of fermentation medium significantly enhanced amylase production (24.65 U/ml) in relation to other test waters at P>0.05. Comparatively, there was a significant increase in amylase produced under optimized condition than not. Conclusively, the obtained results which revealed high amylase yield with available and cost effective agro-wastes is promising for industrial application in Nigeria.},
     year = {2022}
    }
    

    Copy | Download

  • TY  - JOUR
    T1  - Optimization of Amylase Production from Lactobacillus plantarum CS by Submerged Fermentation Using Agro Wastes Substrates
    AU  - George-Okafor Uzoamaka Ogechi
    AU  - Nwachukwu Ujunwa Felicia
    AU  - Ezeme-Nwafor Amara
    Y1  - 2022/02/09
    PY  - 2022
    N1  - https://doi.org/10.11648/j.abb.20221001.12
    DO  - 10.11648/j.abb.20221001.12
    T2  - Advances in Bioscience and Bioengineering
    JF  - Advances in Bioscience and Bioengineering
    JO  - Advances in Bioscience and Bioengineering
    SP  - 11
    EP  - 18
    PB  - Science Publishing Group
    SN  - 2330-4162
    UR  - https://doi.org/10.11648/j.abb.20221001.12
    AB  - Amylases are known to be of great importance worldwide. Most of their production using refined carbon and nitrogen substrates are expensive, resulting in the production of expensive goods which are not easily affordable by end users in our country Nigeria. This necessitates the call for the use of cost-effective substrates under optimized condition for maximum amylase yield. Hence, the present study was focused on the optimization of amylase production from Lactobacillus plantarum CS using agro wastes from local substrates. The parameters assayed were carbon and nitrogen substrates from agro wastes, fermentation period (24-168h), pH (3.5-11) and temperature (30°C-70°C). Others included solvents for preparation of fermentation medium and metal ions of Mg2+, Mn2+, K+, Na+, Hg2+, Fe2+, Cu2+ and Pb2+ as supplements. The amylase produced via shake-flask fermentation was assayed using Dinitro salicylic acid method. The result revealed 3% sweet potato and 1% bambara nut wastes as the best carbon and nitrogen components of the required fermentation medium of pH 6.5. The optimal amylase produced (38.6U/ml) was achieved within 48h at 35°C with 1% inoculum and Mn2+/ Mg2+ as a co-factor. Relatively, Cu2+, Pb2+ and Hg2+ reduced the amylase yield to 2+ and Mg2+ which enhanced the production to 121.66% and 101.09% respectively. Natural rain water used as solvent for the preparation of fermentation medium significantly enhanced amylase production (24.65 U/ml) in relation to other test waters at P>0.05. Comparatively, there was a significant increase in amylase produced under optimized condition than not. Conclusively, the obtained results which revealed high amylase yield with available and cost effective agro-wastes is promising for industrial application in Nigeria.
    VL  - 10
    IS  - 1
    ER  - 

    Copy | Download

Author Information
  • Department of Applied Microbiology and Brewing, Faculty of Applied Natural Sciences, Enugu State University of Science and Technology, Agbani, Nigeria

  • Department of Applied Microbiology and Brewing, Faculty of Applied Natural Sciences, Enugu State University of Science and Technology, Agbani, Nigeria

  • Department of Applied Microbiology and Brewing, Faculty of Applied Natural Sciences, Enugu State University of Science and Technology, Agbani, Nigeria

  • Sections