INHIBIION ACTIVITY OF SOME PLANT SEEDS EXTRACTS AGAINST THE PROTEASE PRODUCED FROM LOCAL ISOLATE OF CANDIDA ALBICANS ISOLATE USING SOLID STATE FERMENTATION
DOI:
https://doi.org/10.36103/dz6j4z41Keywords:
C. albicans ,protease ,inhibitors, plant extract, seedsAbstract
This study was aimed at inhibition of purified protease produced by C. albicans using plant seeds extract. One hundred two local C. albicans isolates that were isolated and identified by microscopic and biochemical tests were submitted to primary and secondary screening techniques in order to select the qualified C. albicans isolate for protease synthesis. Among these isolates, nineteen isolates with the highest hydrolysis zone on skim milk media (primary screening) were chosen for secondary screening. It has been found that C. albicansV56 had the highest productivity of the enzyme (41.8U/mg protein). The optimum conditions of protease production by selected isolate using solid state fermentation by using wheat bran as best substrate , temperature 28 °C and pH 7, after 5 days of incubation . The enzyme was purified by concentration with sucrose, then used gel filtration chromatography using Sephadex G-75. The results show two peaks. The first peak has a purification folds 6.5 time with an enzyme yield of 13.08 %, while peak 2 has a purification folds 3.3 time with an enzyme yield of 20%. The purified enzyme exhibited maximal activity at pH 6.0 for peak 1 and 3.0 for peak 2, whereas the maximum stability was 7.0 for peak 1 and 8 for peak 2. The optimal temperature for purified enzyme activity was 40 for peak 1 and 30 for peak 2, and it was stable until 45°C. Protease activity was inhibited with local plant seed extracts. Lathyrus sativus extract inhibits approximately 50 % of protease activity.
References
Abbas L., A. Hashim and B. Khadhim, 2020. Evaluation of myrtus communis flavonoid as antidermatophtic and keratinase inhibitor. Iraq J Agri Sci, 51(6):1525-1533
BenjakulS; W; and Thummaratwasik P., 2000. Isolation and characterization of trypsin inhibitors from some Thai legume seeds, J. Food Biotch.24;107-127.
Bradford, M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Analytical Biochemistry Journal. 72: 248-254
Bulet P; R. Stocklin; and L. Menin, 2004. Anti-microbial peptide :from invertebrate to vertebrate . Immunol .Rev.198:169-184.
Bull, A.T. and M.E. Bushnell, 1976 Environmental control of fungal growth. In: The filamentous fungo. (ed. Smith, J.E. and Berry, D.R.). 2: 1-29. Edward Arnold. London. (700N).
Castro, A. M., Andrea, T. W., Carvalho, D. F., Teixeira, M. M. P., Castilho, L. R., & Freire, D. M. G., 2011. Production of lipase and protease by Candida utilis in solid state fermentation. Waste Biomass Valor, 2, 291–302.(1500N).
Chang-H, E., Kang, M. S. and J.-J. Kim. 2020. Elastase/Collagenase Inhibition composition of Citrus unshiu and its association with phenolic content and ant-Oxidant actvity. Journal Applied Science. 10N
Chesworth, J. M., Stuchbury, T. and Scaif, J. R., 1998. An Introduction to agricultural biochemistry. Chapman and Hall, London. 5(2): 215-223.
Chisti Y, 1999. Solid substrate fermentations, enzyme production, food enrichment. In Encyclopedia of Bioprocess Technology: Fermentation, Biocatalysis, and Bioseparation, Vol 5, ed. by Flickinger MC and Drew SW. Wiley, New York, pp. 2446–2462
Clive, D., 2002. A guide to protein isolation. Kluwer academic publisher. NewYork.
Dennis, E. A., Cao, J., Hsu, Y. H., Magrioti, V., and Kokotos, G. 2011. Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chemical reviews. 111(10): 6130-6185
Fullbrook, P. D., 1983. The use of enzymes in the processing of oilseeds." Journal of the American Oil Chemists, Society 60, no. 2 (1983): 476-478
Ghannoum M. A, 2000. Potential role of phospholipases in virulence and fungal pathogenesis. Clinical Microbiol Rev.13:122–43
Graf, K., Last, A., Gratz, R., Allert, S., Linde, S., Westermann, M., Groger, M., Mosig, A.S., Gresnigt, M.S., and Hube, B. 2019. Keeping Candida albicans commensal: how lactobacilli antagonize pathogenicity of Candida albicans in an in vitro gut model. Dis Model Mechanisms. 12(9):39719.
Kim J.Y, S.C Park, Lee J.K, Hwang I, Cheong H, Nah J.W, Hahm K.S, Park Y . 2009. Antifungal mechanism of a novel antifungal protein from pumpkin rinds against various fungal pathogens. J Agric Food Chem 57:9299–9304
Kocher and Mishra, 2009. Immobilization of Bacillus circulans MTCC 7906 for enhanced production of alkaline protease under batch and packed bed fermentation. Internet J. Microbial, 7: 359-378.
Maher, H. R., and Cordes, E. A. 1971. Biochemistry of bacterial growth. 3rd ed. Blackwell Scientific publication, Oxford
Mohanasrinivasan V, Shankar V, Elizabeth R, Soumya A. R, Devi C. S 2012. Isolation, screening and identification of protease producing fungi from rhizosphere soil and optimization of pH, incubation time and inducer concentration for enhanced protease production. Int. J. Pharm. Biol. Sci., 3(2): 784-793
Mona M. Rasheed., E. Abeer Mahmoud, S. Amir Al-Kashef and U. Mohammed Nooman, 2012. Purification and Characterization of a Novel Fibrinolytic Enzyme by Candida guilliermondii Grown on Sunflower Oil Cake. Journal of Applied Sciences Research, 8(2): 635-645, 2012
Park, M.-H., Walpola B. C. and M.-H. Yoon. 2013. Purification and characterization of protease enzyme from Burkholderiastabilis. African Journal of Biotechnology 12(12).
Phadatare S.U., Deshpande, V.V. and Srinivasan, M.C. (1993). High activity alkaline protease from Condiobolus coronatus .Enzyme MicrobTechnol, 15: 72-76
Sabri.C.A , Hakeem.M.I , 2016 . Purification and characterization of salivary amylase inhibitor extracted from barly.Iraq J Agri Sci, 47(4):1039-1048
Sahar. I. H, A. F. Khalaf, Y. Ahmed B. Ahmed and A.Iyad,2020.Determination of inhibition activity of α- amylase enzyme, antioxidant activity, anti-bacteria and phenolic compound by using some of medical plant.Iraq J Agri Sci,51(1):411-421
Saleh, G. M. 2020. Isolation and characterization of unique fructophilic lactic acid bacteria from different flower sources. Iraqi Journal of Agricultural Sciences, 51(2), 508-518. https://doi.org/10.36103/ijas.v51i2.977
Segel, J. J. (1976). Biochemical Calculation. John Wiley and sons.(170N).
Smitha B.R and R. Shashanka, 2011. Isolation and identification of candida from oral cavity. ISNR Dentistry, Vol 487921,7 pages
Sugita T, Kurosaka S, Yajitate M, Sato H, Nishikawa A. 2002. Extra cellular proteinase and phospholipase activity of three genotypic strains of a human pathogenic yeast, Candida albicans. MicrobiolImmunol. 2002;46:881
Supuran, C. T., Scozzafava, A. and B. W. Clare. Bacterial protease inhibitors. Medicinal Research Reviews. 22 (4): 329-372.
Sutar I.I., M. C. Srinivasan & H. G. Vartak, 1992. Production of an alkaline proteinase from Conidiobolus coronatus and its use to resolve DL-phenylalanine and DL-phenylglycine. World Journal of Microbiology and Biotechnology volume 8, pp: 254–258
Wibawa, T. The role of virulence factors in Candida albicans pathogenicity. J. Med. Sci. 2016, 48, 58–68
Wu L. C. and Y. D. Hang, Letters in Applied Microbioliology, 1998. Pages: 201
Yakkou, L., S. Houida M Raouane, A., El Harti, and S. Amghar, 2022. Effect of earthworm's extracts (Aporrectodea molleri) on yeast growth: in vitro study. Iraqi Journal of Agricultural Sciences, 53(1):173-179. https://doi.org/10.36103/ijas.v53i1.1522
Yonis, R.W., K. J Luti, and G. M., Aziz, 2019. Statistical optimization of chitin bioconversion to produce an effective chitosan in solid state fermentation by Aspergillus flavus. Iraqi Journal of Agricultural Sciences, 50(3);916-927. https://doi.org/10.36103/ijas.v50i3.708
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