Phytase Production from a Novel Klebsiella sp. on Wheat Bran for Animal Feed Digestion
Dilruba Akter a, Md. Murad Khan b, Md. Mahmuduzzaman Mian a, Shakila Nargis Khan a, Md. Mozammel Hoq a
Microbial Bioactives 1(1) 014-021 https://doi.org/10.25163/microbbioacts.11004A0423100718
Submitted: 03 March 2018 Revised: 23 May 2018 Published: 02 April 2019
Abstract
Background: The present study was aimed to isolate phytase producing bacteria and optimize the physicochemical parameters of their phytase production. Materials and methods: Four bacterial isolates (Phs4, Phs5, Phs6, and Phs8), based on clear zone formation on phytase screening medium, were selected and tested for finding out the highest phytase producing strain. The production of phytase was then optimized and its biochemical properties were determined to judge the applicability of phytase as a digestive aid in animal feed. Results: The 4 bacterial isolates (Phs4, Phs5, Phs6 and Phs8) were identified by morphological, cultural, biochemical and molecular characterization as Burkholderia cepacia, Escherichia coli, Klebsiella pneumoniae and Klebsiella sp. respectively. Of these isolates, Phs8 (Klebsiella sp.) was found to produce maximum phytase in shake culture in a basal medium containing Na-phytate at 37oC and pH 5.5 after 72 hours of incubation. The omission of Na-phytate from the medium almost completely abolished the phytase production capacity of the isolate and thus signified its important role as an inducer. Among the different complex carbon sources, viz., glucose, wheat bran, rice bran and chickpea, maximum phytase production (94 unit/ ml) was obtained with wheat bran under comparable cultivation conditions. The phytase works best at a temperature of 37oC and pH of 4.0 with a wide temperature stability (more than 80% activity up to 80oC) and wide pH stability (more than 80% activity within a range of 3-8). Although Zn2+, Co2+, and Fe2+ slightly increased the phytase activity Cu2+ and Mg2+ strongly inhibited the enzyme. Conclusion: The present findings will be very useful for the development of a bioprocess of the enzyme for its large-scale production at the pilot and finally at the commercial level.
Keywords: Phytase; animal feed; Klebsiella sp.; enzyme activity.
Significance: Economic production of bacterial phytase as a poultry feed supplement
Abbreviations: PSM, phytase screening medium; PPM, phytase production medium; LB, Luria-Bertani; RPM, rotation per minute; PCR, polymerase chain reaction; dNTPs, deoxyribonucleotides; EDTA, ethylenediaminetetraacetic acid; EtBr, ethidium bromide; KIA, Kligler iron agar; MIU, motility indole urease test; VP, Voges–Proskauer test.
References
Aly, M. M., Tork, S., Al-Garni, S. M., & Kabli, S. A. (2015). Production and Characterization of Phytase from Streptomyces luteogriseus R10 Isolated from Decaying Wood Samples. International Journal of Agriculture and Biology, 17(3), 515-522. doi:http://dx.doi.org/10.17957/ijab/17.3.14.453
Badone, F. C., Amelotti, M., Cassani, E., & Pilu, R. (2012). Study of low phytic acid1-7 (lpa1-7), a new ZmMRP4 mutation in maize. Journal of Heredity, 103(4), 598-605. doi:http://dx.doi.org/10.1093/jhered/ess014
Bae, H., Yanke, L., Cheng, K.-J., & Selinger, L. (1999). A novel staining method for detecting phytase activity. Journal of Microbiological Methods, 39(1), 17-22. doi:http://dx.doi.org/10.1016/s0167-7012(99)00096-2
Bilgiçli, N., & Ibanoglu, S. (2007). Effect of wheat germ and wheat bran on the fermentation activity, phytic acid content and colour of tarhana, a wheat flour–yoghurt mixture. Journal of Food Engineering, 78(2), 681-686. doi:https://doi.org/10.1016/j.jfoodeng.2005.11.012
Cho, J. S., Lee, C. W., Kang, S. H., Lee, J. C., Bok, J. D., Moon, Y. S., . . . Choi, Y. J. (2003). Purification and characterization of a phytase from Pseudomonas syringae MOK1. Current Microbiology, 47(4), 0290-0294. doi:http://dx.doi.org/10.1007/s00284-002-3966-4
Elkhalil, E., Männer, K., Borriss, R., & Simon, O. (2007). In vitro and in vivo characteristics of bacterial phytases and their efficacy in broiler chickens. British Poultry Science, 48(1), 64-70. doi:http://dx.doi.org/10.1080/00071660601148195
Escobin-Mopera, L., Ohtani, M., Sekiguchi, S., Sone, T., Abe, A., Tanaka, M., . . . Asano, K. (2012). Purification and characterization of phytase from Klebsiella pneumoniae 9-3B. Journal of Bioscience and Bioengineering, 113(5), 562-567. doi:http://dx.doi.org/10.1016/j.jbiosc.2011.12.010
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39(4), 783-791. doi:https://doi.org/10.1111/j.1558-5646.1985.tb00420.x
Greiner, R., & Carlsson, N.-G. (2006). Myo-Inositol phosphate isomers generated by the action of a phytate-degrading enzyme from Klebsiella terrigena on phytate. Canadian Journal of Microbiology, 52(8), 759-768. doi:http://dx.doi.org/10.1139/w06-028
Greiner, R., Konietzny, U., & Jany, K.-D. (1993). Purification and characterization of two phytases from Escherichia coli. Archives of Biochemistry and Biophysics, 303(1), 107-113. doi:http://dx.doi.org/10.1006/abbi.1993.1261
Gupta, R. K., Gangoliya, S. S., & Singh, N. K. (2015). Reduction of phytic acid and enhancement of bioavailable micronutrients in food grains. Journal of Food Science and Technology, 52(2), 676-684. doi:http://dx.doi.org/10.1007/s13197-013-0978-y
Harland, B. F., & Morris, E. R. (1995). Phytate: a good or a bad food component? Nutrition Research, 15(5), 733-754. doi:http://dx.doi.org/10.1016/0271-5317(95)00040-p
Haros, M., Bielecka, M., Honke, J., & Sanz, Y. (2007). Myo-inositol hexakisphosphate degradation by Bifidobacterium infantis ATCC 15697. International Journal of Food Microbiology, 117(1), 76-84. doi:http://dx.doi.org/10.1016/j.ijfoodmicro.2007.02.021
Hegsted, D. (1968). Present knowledge of calcium, phosphorus, and magnesium. Nutrition Reviews, 26(3), 65-70. doi:http://dx.doi.org/10.1111/j.1753-4887.1968.tb00862.x
Hill, J. E., Kysela, D., & Elimelech, M. (2007). Isolation and assessment of phytate-hydrolysing bacteria from the DelMarVa Peninsula. Environmental Microbiology, 9(12), 3100-3107. doi:http://dx.doi.org/10.1111/j.1462-2920.2007.01420.x
Hong, S. W., Chu, I. H., & Chung, K. S. (2011). Purification and biochemical characterization of thermostable phytase from newly isolated Bacillus subtilis CF92. Journal of the Korean Society for Applied Biological Chemistry, 54(1), 89-94. doi:http://dx.doi.org/10.3839/jksabc.2011.012
Jones, G. (2013). How to select the best phytase for your feed formulation. Retrieved from https://www.wattagnet.com/articles/17645-how-to-select-the-best-phytase-for-your-feed-formulation
Jorquera, M., MARTíNEZ, O., Maruyama, F., Marschner, P., & de la Luz Mora, M. (2008). Current and future biotechnological applications of bacterial phytases and phytase-producing bacteria. Microbes and Environments, 23(3), 182-191. doi:http://dx.doi.org/10.1264/jsme2.23.182
Kalsi, H. K., Singh, R., Dhaliwal, H. S., & Kumar, V. (2016). Phytases from Enterobacter and Serratia species with desirable characteristics for food and feed applications. 3 Biotech, 6(1), 1-13. doi:http://dx.doi.org/10.1007/s13205-016-0378-x
Kerovuo, J., & Tynkkynen, S. (2000). Expression of Bacillus subtilis phytase in Lactobacillus plantarum 755. Letters in Applied Microbiology, 30(4), 325-329. doi:http://dx.doi.org/10.1046/j.1472-765x.2000.00660.x
Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16(2), 111-120. doi:https://doi.org/10.1007/BF01731581
Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Molecular Biology and Evolution, 35(6), 1547-1549. doi:https://doi.org/10.1093/molbev/msy096
Maathuis, F. J. (2009). Physiological functions of mineral macronutrients. Current Opinion in Plant Biology, 12(3), 250-258. doi:http://dx.doi.org/10.1016/j.pbi.2009.04.003
Mullaney, E. J., & Ullah, A. H. (2003). The term phytase comprises several different classes of enzymes. Biochemical and Biophysical Research Communications, 312(1), 179-184. doi:http://dx.doi.org/10.1016/j.bbrc.2003.09.176
Pandey, A., Szakacs, G., Soccol, C. R., Rodriguez-Leon, J. A., & Soccol, V. T. (2001). Production, purification and properties of microbial phytases. Bioresource Technology, 77(3), 203-214. doi:http://dx.doi.org/10.1016/s0960-8524(00)00139-5
Quan, C.-S., Tian, W.-J., Fan, S.-D., & Kikuchi, J.-I. (2004). Purification and properties of a low-molecular-weight phytase from Cladosporium sp. FP-1. Journal of Bioscience and Bioengineering, 97(4), 260-266. doi:http://dx.doi.org/10.1016/s1389-1723(04)70201-7
Saitou, N., & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4(4), 406-425. doi:https://doi.org/10.1093/oxfordjournals.molbev.a040454
Sajidan, A., Farouk, A., Greiner, R., Jungblut, P., Müller, E.-C., & Borriss, R. (2004). Molecular and physiological characterisation of a 3-phytase from soil bacterium Klebsiella sp. ASR1. Applied Microbiology and Biotechnology, 65(1), 110-118. doi:http://dx.doi.org/10.1007/s00253-003-1530-1
Selle, P. H., Cowieson, A. J., Cowieson, N. P., & Ravindran, V. (2012). Protein–phytate interactions in pig and poultry nutrition: a reappraisal. Nutrition Research Reviews, 25(1), 1-17. doi:http://dx.doi.org/10.1017/s0954422411000151
Soni, S. K., & Khire, J. M. (2007). Production and partial characterization of two types of phytase from Aspergillus niger NCIM 563 under submerged fermentation conditions. World Journal of Microbiology and Biotechnology, 23(11), 1585-1593. doi:http://dx.doi.org/10.1007/s11274-007-9404-9
Spier, M. R., Greiner, R., Rodriguez-León, J. A., Woiciechowski, A. L., Pandey, A., Soccol, V. T., & Soccol, C. R. (2008). Phytase production using citric pulp and other residues of the agroindustry in SSF by fungal isolates. Food Technology & Biotechnology, 46(2), 178-182. doi:http://dx.doi.org/10.1080/00986445.2010.493115
Unno, Y., Okubo, K., Wasaki, J., Shinano, T., & Osaki, M. (2005). Plant growth promotion abilities and microscale bacterial dynamics in the rhizosphere of Lupin analysed by phytate utilization ability. Environmental Microbiology, 7(3), 396-404. doi:http://dx.doi.org/10.1111/j.1462-2920.2004.00701.x
Wang, X., Upatham, S., Panbangred, W., Isarangkul, D., Summpunn, P., Wiyakrutta, S., & Meevootisom, V. (2004). Purification, characterization, gene cloning and sequence analysis of a phytase from Klebsiella pneumoniae subsp. pneumoniae XY-5. Science Asia, 30, 383-390. doi:http://dx.doi.org/10.2306/scienceasia1513-1874.2004.30.383
Zhang, R., Yang, P., Huang, H., Shi, P., Yuan, T., & Yao, B. (2011). Two types of phytases (histidine acid phytase and β-propeller phytase) in Serratia sp. TN49 from the gut of Batocera horsfieldi (Coleoptera) larvae. Current Microbiology, 63(5), 408. doi:http://dx.doi.org/10.1007/s00284-011-9995-0
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