Production of L-lysine under submerged fermentation by Corynebacterium glutamicum using different plants' leaves

Authors

  • Theresa EZEDOM Delta State University, Abraka, Nigeria Author https://orcid.org/0000-0002-3942-923X
  • Egoamaka Oliseneku Egbune Author
  • Solomon Adanoritsewo Atseponu Author
  • Mary Ogochukwu Charles Author
  • Blessed Achugbue Benson Author
  • Diana Ebbah Author
  • Promise Chika Amechi Author
  • Oghenetega Benjamin Author
  • Akpevweoghene Rejoice Egbodje Author
  • Lucky Ebinum Author
  • Blessing Ifechi Chukwudozie Author
  • Stephen Eboe Author
  • Ifeanyi Benedict Alexander Author
  • Sophia Fejiro Edijana Author
  • Nyerhovwo Tonukari Author https://orcid.org/0009-0000-0049-8783

Keywords:

Agricultural leaves, submerged fermentation, Corynebacterium glutamicum, extract, boiling

Abstract

Introduction: This study investigated the production of L-lysine using Corynebacterium glutamicum and
various leaf extracts (cassava, palm tree, maize, cowpea, cocoyam, and plantain). The study also explored
the activities of amylases and proteases, as well as the levels of total soluble proteins, reducing sugars,
glucose, flavonoid and phenolic contents, and pH changes.

Materials and Methods: Different treatments (extract, boiled extract, extract + C. glutamicum, boiled
extract + C. glutamicum) were examined for their effects on L-lysine concentration. Additionally, the
activities of amylases and proteases, as well as levels of total soluble proteins, reducing sugars, glucose,
flavonoid and phenolic contents, and pH changes, were analyzed.

Results: Maize leaf extract + C. glutamicum exhibited the highest L-lysine concentration (1.771a±0.1
mg/g), while boiled cassava leaf extract showed the lowest concentration (0.023b±0.1 mg/g). Palm tree
leaf extract had significantly higher reducing sugar levels compared to other extracts. Boiled plantain leaf
extract fermented by C. glutamicum had the highest total soluble protein level (9.5±0.2 mg/g), while
cassava leaf extract had the lowest (2.1±1.2 mg/g).

Conclusion: Submerged fermentation of leaf extracts using C. glutamicum can be utilized for L-lysine
production. The study highlights the influence of different leaf extracts and treatments on L-lysine
production, as well as on amylase and protease activities, total soluble protein levels, reducing sugars,
glucose, flavonoid and phenolic contents, and pH values. These findings provide valuable insights into
the potential application of this approach for lysine production.

References

Anastassiadis, S. (2007). L-lysine fermentation. Resent Patents Biotechnol. 1: 11-24.

Egbune, E. O, Eze, E, Edwards, R. A., Ezedom, T. and Tonukari NJ, (2021b). Enhancement of the nutritional value of elephant grass (PennisetumpurpureumSchum.) for use as animal feeds and for xylanase production. Nigerian Journal of Science and Environment, 19(2).

Ojo, I., Apiamu, A., Egbune, E. O. and Tonukari, N. J. (2022).Biochemical Characterization of Solid-State Fermented Cassava Stem (ManihotesculentaCrantz-MEC) and Its Application in Poultry Feed Formulation.Applied Biochemistry and Biotechnology, 194(6), 2620-2631.

Nelofer, R., Syed Q. and Nadeem M. (2008). Statistical Optimization of Process Variables for L-lysine production by Corynebacteriumglutamicum. Turk. J. Biochem., 33(2): 50-57.

Shah H., Hameed A., A., and Majidkhan G. (2002). Fermentative production of L-lysine:C. glutamicuml fermentation-I. J. Med. Sci. 2: 52-157.

Nelofer R, Ramanan RN, Rahman RN, Basri M, Ariff AB (2012) Comparison of the estimation capabilities of response surface methodology and artificial neural network for the optimization of recombinant lipase production by E. coli BL21. J IndMicrobiolBiotechnol 39:243–254. doi:10.1007/s10295-011-1019-3.

Ekwealor, I.A. and Obeta, J.A.N. (2005) Studies on Lysine Production by Bacillus megaterium. African Journal of Biotechnology, 4, 633-638.

http://dx.doi.org/10.5897/AJB2005.000-3115

Ikeda, M. (2003).Amino acid production process. Adv. Biochem. Eng. Biotechnol. 79: 1–35.

Kircher, M., and Pfeerle W. (2001). The fermentative production of L-lysine as an animal feed additive. Chemosphere. 43: 27-31.

Seibold, G., M., Auchter, S. Berens, J. Kalinowski and B.J. Eikmanns. (2006). Utilization of soluble starch by a recombinant Corynebacteriumglutamicum strain: growth and lysine production. J. Biotechnol., 124: 381-391.

Coello, N., Brito L. and Nonus M. (2000). Biosynthesis of L-lysine by Corynebacteriumglutamicumgrown on fish silage. Bioresource Technol. 73: 221-225.

Chinard, F.D. (1952) Photometric Estimation of Proline and Ornithine. Journal of Biological Chemistry, 199, 91-95.

Gornall, A., Bardsmill, C. T., David, M. M. (1949). Determination of protein by means of biuret reaction. J. Biol. Chem. 177: 751-766.

Miller GL, 1959. Use of the Dinitrosalicylic Acid Reagent for the Determination of Reducing Sugar.Anal. Chem. 31 426-428.

Nouadri, T., Meraihi, Z., Shahrazes, D. D. and Leila, B. (2010). Purification and characterization of amylase isolated from Penicilliumcamemberti PL21. African Journal of Biochemistry Research 4 (6): 155-162.

Kunitz M. (1947). “Crystalline soybean trypsin inhibitor. II. General properties,” The Journal of General Physiology, 30, 4, 291–310.

Jia Z, Tang M, Wu J. (1999). The determination of flavonoid contents of murlberry and their scavenging effects on superoxide radicals. Food Chemistry, 64, 555-559.

Singleton VL, Rossi JA (1965).Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture 16(3):144-158.

Teniola O.D. and Odunfa S. A. (2001). The effects of processing methods on the levels of lysine, methionine and the general acceptability of ogi processed using starter cultures. Int J Food Microbiol . 2001 Jan 22;63(1-2):1-9. doi: 10.1016/s0168-1605(00)00321-4.

Cui Li, Li D. and Liu C. (2012).Effect of fermentation on the nutritive value of maize. International Journal of Food Science and Technology 2012, 47, 755–760.

Ezemba, C.C., Ozokpo, C.A., Anakwenze, V.N., Anaukwu, G.C., Ogbukagu, C.M., Ekwealor, C.C. and Ekwealor, I.A. (2016) Lysine Production of Microbacteriumlacticum by SubmergedFermentation Using Various Hydrocarbon, Sugar and Nitrogen Sources. Advances in Microbiology , 6, 797-810

Amaechi N. C. and Oluagha N. E. (2016).effect of boiling on amino acid composition ofjackfruit (Artocarpusheterophyllus) seed from south-East Nigeria.

Hurrell R.F. and Finot P.A. (1983).Food processing and storage as a determinant of protein and amino acid availability.Experientia ;Suppl. 44: 135-156.

Gersten D. (2013). The 20 amino acids: What they do and how they keep you alive and vibrant. The Gersten Institute for Higher Medicine.San Diego, California, USA; 2013.

Hsu, C.B., Cheng, S.P., Hsu, J.C., Yen, H.T., 2001. Effect of threonine addition to a low protein diet on IgG levels in body fluid of first-litter sows and their piglets. Asian-Australasian Journal of Animal Sciences 14, 1157–1163.

Zhu, Y.P., Fan, J.F., Cheng, Y.Q., Li, L.T., 2008. Improvement of the antioxidant activity of Chinese traditional fermented okara (Meitauza) using Bacillus subtilis B2. Food Control 19, 654–661.

Ezekiel O.O., Aworh O. C., Blaschek H. P., and Thaddeus C. (2010).Protein enrichment of cassava peel by submerged fermentation withTrichodermaviride (ATCC 36316).African Journal of Biotechnology Vol. 9 (2), pp. 187-194.

Egbune E. O., Orhonigbe I., Adheigu R. O., Oniyan U. P. and Tonukari N. J. (2021a). Effect of inoculum size on solid state fermentation of pearl millet (Pennisetumglaucum) by Rhizopusoligosporus. Nigerian Journal of Science and Environment 20 (1): 1-9.

Ezedom, T., Egbune, E., Ehikordi, M., Ezeugo, N., Eledu, F., Esiete, J., Eriamiator, J., Ezeugo, O., Friday, C., Egholenwa, C., Ekene, E., Efemevotor, S., &Tonukari, N. (2022). Biochemical evaluation of autoclaved and solid state fermented tropical pasture grasses. Journal of Agricultural Biotechnology and Sustainable Development, 14(2), 24-32.

Vijayaraghavan P, Lourthuraj AA, Arasu MV, Al-Dhabi NA, Ravindran B, Woong CS., (2021). Effective removal of pharmaceutical impurities and nutrients using biocatalyst from the municipal wastewater with moving bed packed reactor, Environmental Research,Volume 200,111777,ISSN 0013-9351, https://doi.org/10.1016/j.envres.2021.111777.

Ray, A. K., Ghosh, K., and Ringø, E. (2012). Enzyme-producing C. glutamicum isolated from fish gut: a review. Aquaculture Nutrition, 18(5), 465–492. doi:10.1111/j.1365-2095.2012.00943.

Wei S, Lu G, Cao H. (2017). Effects of cooking methods on starch and sugar composition of sweetpotato storage roots. PLoS One. Aug 21;12(8):e0182604. doi: 10.1371/journal.pone.0182604. PMID: 28827808; PMCID: PMC5565179.

Lai Y.C., Huang C.L., Chan C.F., Lien C.Y. and Liao WC (2013).Studies of sugar composition and starch morphology of baked sweet potatoes (Ipomoea batatas (L.) Lam). J Food SciTechnol 50: 1193–1199. doi: 10.1007/s13197-011-0453-6.

Bian K and Liu X (2012) Determination of soluble suagrs by HPLC and their changes duing processing in sweet potato. J Henan UnivTechnol (Nat Sci Ed) 33: 1–5.

Lu G, Jiang Y, Wu J (1997) Change of starch and sugar of sweet potato storage roots during processing. ActaAgric Zhejiang 1997: 78–82.

Nkhata SG, Ayua E, Kamau EH, Shingiro JB. (2018). Fermentation and germination improve nutritional value of cereals and legumes through activation of endogenous enzymes. Food SciNutr. 16;6(8):2446-2458. doi: 10.1002/fsn3.846. PMID: 30510746; PMCID: PMC6261201.

Osman, M. A. (2011). Effect of traditional fermentation process on the nutrient and antinutrient contents of pearl millet during preparation of Lohoh. Journal of the Saudi Society of Agricultural Sciences, 10, 1–6.

El‐Hag, M. E. , El‐Tinay, A. H. andYousif, N. E. (2002). Effect of fermentation and dehulling on starch, total polyphenols, phytic acid content and in vitro protein digestibility of pearl millet. Food Chemistry, 77, 193–196. 10.1016/S0308-8146(01)00336-3.

Anigboro, A. A., Aganbi, E. and Tonukari, N. J. (2020).Solid State Fermentation of Maize (Zea mays) Offal by Rhizopusoligosporusunder Acidic and Basic Conditions.J. Sci. R., 12 (4): 751 – 756.

Kathiresan K. and Manivannan S. (2006).α Amylase production by Penicilliumfellutanum isolated from mangrove rhizosphere soil. African J. Biotech. 2006;5(10):829–832.

Egbune, E. O., Avwioroko, O. J., Anigboro A. A., Aganbi, E., Amata, A., Tonukari, N. J. (2022). Characterization of a surfactant-stable α-amylase produced by solid-state fermentation of cassava (ManihotesculentaCrantz) tubers using Rhizopusoligosporus: Kinetics, thermal inactivation thermodynamics and potential application in laundry industries. Biocatalysis and Agricultural Biotechnology 39. 102290

Dou, S., Chi, N., Zhou, X., Zhang, Q., Pang, F., Xiu, Z. (2018). Molecular cloning, expression, and biochemical characterization of a novel cold-active α-amylase from Bacillus sp. dsh19-1. Extremophiles 22, 739–749

Divakaran, D., Chandran, A., PratapChandran, R., 2011. Comparative study on production of α-amylase from Bacillus licheniformis strains. Braz. J. Microbiol. 42, 1397–1404

Anto H., Trivedi U. and Patel K. (2006). Alpha Amylase Production by Bacillus cereusMTCC 1305 Using Solid-State Fermentation. s, Food Technol. Biotechnol. 44 (2) 241–245.

Saxena R. and Singh R. (2011). Amylase production by solid-state fermentation of agro-industrial wastes using Bacillus sp. Braz J Microbiol. 2011 Oct;42(4):1334-42. doi: 10.1590/S1517-838220110004000014. Epub 2011 Dec 1. PMID: 24031761; PMCID: PMC3768732.

Punniyakotti, E., Lim, J., Park, Y., Cho, M., Shea, P. and Oh, B. (2020). Agricultural waste materials enhance protease production by Bacillus subtilis B22 in submerged fermentation under blue light-emitting diodes. Bioprocess and Biosystems Engineering. 43. 10.1007/s00449-019-02277-5.

Mathias TRS, Mello PPM, Servulo EFC (2014) Solid wastes in brewing process: a review. J Brew Distilling 5(1):1–9. https://doi.org/10. 5897/JBD2014.0043

Asoodeh A, Emtenani S, Emtenani S, Jalal R (2014). Enzymatic Molecular cloning and biochemical characterization of a thermoacidophilic, organic-solvent tolerant a-amylase from a Bacillus strain in Escherichia coli. J MolCatal B Enzym 99:114–120. https ://doi.org/10.1016/j.molcatb.2013.10.025

Farhadian S, Asoodeh A, Lagzian M (2015). Purifcation, biochemical characterization and structural modeling of a potential htrA-like serine protease from Bacillus subtilis DR8806. JMolCatal B Enzym 115:51–58. https://doi.org/10.1016/j.molca tb.2015.02.001

Adetuyi F.O. and Ibrahim T.A (2014).Effect of fermentation time on the phenolic, flavonoid and vitamin c contents and antioxidant activities of okra (Abelmoschusesculentus) seeds. Nigerian Food Journal Vol. 32 No. 2, pages 128 – 137.

Yao Q., Xiao-Nan J., Dong P.H. (2010). Comparison of antioxidant activities in black soybean preparations fermented with various microorganisms. Agricultural Science in China, 9 (2010), pp. 1065-1071.

Moktan, B., Saha, J. and Sarkar, P.K. (2008).Antioxidant activities of soybean as affected by Bacillus fermentation to kinema. Food Research International 41: 586 – 593.

Ademiluyi A.O. and Oboh G. (2011). Antioxidant properties of condiment produced from fermented bambara groundnut (Vigna subterranean L. Verdc). Journal of Food Biochemistry, 35 (2011), pp. 1145-1160.

James, S., Nwabueze T. U., Ndife J., Onwuka G. I., Usman M. A. (2020). Influence of fermentation and germination on some bioactive components of selected lesser legumes indigenous to Nigeria, Journal of Agriculture and Food Research, Volume 2, 2020, 100086,ISSN 2666-1543, https://doi.org/10.1016/j.jafr.2020.100086.

Egbune, E. O., Aganbi, E., Anigboro, A. A., Ezedom, T., Onojakpor, O., Amata, A. I. and Tonukari, N. J. (2023). Biochemical characterization of solid-state fermented cassava roots (ManihotesculentaCrantz) and its application in broiler feed formulation. World Journal of Microbiology and Biotechnology, 39(2), 1-12.

Ndego, A., Ezedom, T., Egbune, E. O. and Tonukari, N. (2023). Biochemical characterization of solid state fermented maize cob (Zea mays) using Rhizopusoligosporusand its application in poultry feed production. International journal of recycling organic waste in agriculture, 12(2), 235-246.

Rahmi, N., Purnama, D., Umar, S., and Eni, H.. (2016). The effect of fermentation on total phenolic, flavonoid and tannin content and its relation to antiC. glutamicuml activity in jaruktigarun (Crataevanurvala, Buch HAM). 23. 309-315.

Ezedom, T., Egbune, E., Ehikordi, M., Ezeugo, N., Eledu, F., Esiete, J., ...&Tonukari, N. (2022). Biochemical evaluation of autoclaved and solid state fermented tropical pasture grasses. Journal of Agricultural Biotechnology and Sustainable Development, 14(2), 24-32.

Duenas M, Hemandez T, Estrella I, Fernandez D. (2009). Germination as a process to increase the polyphenol content and antioxidant activity of lupin seeds (Lupinusangustifolius L.). Food Chemistry 117(4), 599-607. http://dx.doi.org/10.1016/j.foodchem.2009.04.051

Scherer R. and Godoy H.T. (2009). Antioxidant activity index (AAI) by the 2,2-diphenyl-1- picrylhydrazyl method Food Chemistry, 112 , pp. 654-658.

Dajanta K., Janpum, P. and Leksing, W. (2013). Antioxidant capacities, total phenolics and flavonoids in black and yellow soybeans fermented by Bacillus subtilis: A comparative study of Thai fermented soybeans (thuanao). International Food Research Journal. 20. 3125-3132.

Guzmán-Uriarte M.L., Sánchez-Magaña L.M., Angulo-Meza G.Y., Cuevas-Rodríguez E.O., Gutiérrez Dorado R.,, Mora-Rochín S., Milán-Carrillo J., Valdez-Ortiz A., Reyes-Moreno C. (2013). Solid state bioconversion for producing common bean (Phaseolus vulgaris L.) Functional flour with high antioxidant activity and antihypertensive potential Food and Nutrition Sciences, 4 (2013), pp. 480-490.

Shrestha, A.K., Dahal, N.R. and Ndungustse, V. (2010).Bacillus fermentation of soybean: A review. Journal of Food Science and Technology Nepal 6: 1 – 9.

Alireza C. K. (2017). Re: Why is the pH increasing during fed-batch fermentation?.Retrieved from:https://www.researchgate.net/post/Why-is-the-pH-increasing-during-fed-batch-ermentation/58c39675ed99e15e0c6393cf/citation/download.

Egbune E. O., Orhonigbe I., Adheigu R. O., Oniyan U. P. and Tonukari N. J. (2021). Effect of inoculum size on solid state fermentation of pearl millet (Pennisetumglaucum) by Rhizopusoligosporus. Nigerian Journal of Science and Environment, 20 (1): 1-9.

Downloads

Published

06/05/2023

How to Cite

1.
Production of L-lysine under submerged fermentation by Corynebacterium glutamicum using different plants’ leaves. AJTMBR [Internet]. 2023 Jun. 5 [cited 2024 Dec. 21];6(1):12-24. Available from: https://ajtmbr.org.ng/index.php/home/article/view/87