Abstract
Optimization of process parameters for phytase production by Enterobacter sp. ACSS led to a 4.6-fold improvement in submerged fermentation, which was enhanced further in fed-batch fermentation. The purified 62 kDa monomeric phytase was optimally active at pH 2.5 and 60 °C and retained activity over a wide range of temperature (40–80 °C) and pH (2.0–6.0) with a half-life of 11.3 min at 80 °C. The kinetic parameters K m, V max, K cat, and K cat/K m of the pure phytase were 0.21 mM, 131.58 nmol mg−1 s−1, 1.64 × 103 s−1, and 7.81 × 106 M−1 s−1, respectively. The enzyme was fairly stable in the presence of pepsin under physiological conditions. It was stimulated by Ca+2, Mg+2 and Mn+2, but inhibited by Zn+2, Cu+2, Fe+2, Pb+2, Ba+2 and surfactants. The enzyme can be applied in dephytinizing animal feeds, and the baking industry.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Koppelaar RH, Weikard HP (2013) Assessing phosphate rock depletion and phosphorus recycling options. Glob Environ Change 23:1454–1466
Escobin-Mopera L, Ohtani M, Sekiguchi S, Sone T, Abe A, Tanaka M et al (2012) Purification and characterization of phytase from Klebsiella pneumoniae 9-3B. J Biosci Bioeng 113:562–567
Joshi S, Satyanarayana T (2014) Optimization of heterologous expression of the phytase (PPHY) of Pichia anomala in P. pastoris and its applicability in fractionating allergenic glycinin from soy protein. J Ind Microbiol Biotechnol 41:977–987
Kumar A, Chanderman A, Singh S (2015) Microbial production of phytases for combating environmental phosphate pollution and other diverse applications. Crit Rev Environ Sci Technol. doi:10.1080/10643389.2015.1131562
Howson SJ, Davis RP (1983) Production of phytate-hydrolysing enzyme by some fungi. Enzyme Microb Technol 5:377–382
Heinonen JK, Lahti RJ (1981) A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase. Anal Biochem 113:313–317
Singh B, Satyanarayana T (2006) Phytase production by thermophilic mold Sporotrichum thermophile in solid-state fermentation and its application in dephytinization of sesame oil cake. Appl Biochem Biotechnol 133:239–250
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Bae HD, Yanke LJ, Cheng KJ, Selinger LB (1999) A novel staining method for detecting phytase activity. J Microbiol Methods 39:17–22
Bhavsar K, Ravi Kumar V, Khire JM (2011) High level phytase production by Aspergillus niger NCIM 563 in solid state culture: response surface optimization, up-scaling, and its partial characterization. J Indus Microbiol Biotechnol 38:1407–1417
Sapna, Singh B (2013) Improved production of protease-resistant phytase by Aspergillus oryzae and its applicability in the hydrolysis of insoluble phytates. J Ind Microbiol Biotechnol 40:891–899
Gacula MC, Singh J (1984) Statistical methods in food and consumer research. Academic Press, New York
Greiner R (2007) Phytate-degrading enzymes: regulation of synthesis in microorganisms and plants. Inositol phosphates. Linking agriculture and the environment. CAB International, Wallingford
Williams SG (1970) The role of phytic acid in the wheat grain. Plant Physiol 45:376–381
Guo J, Bian YY, Zhu KX, Guo XN, Peng W, Zhou HM (2015) Activation of endogenous phytase and degradation of phytate in wheat bran. J Agric Food Chem 63:1082–1087
Karthic P, Joseph S, Arun N, Kumaravel S (2013) Optimization of biohydrogen production by Enterobacter species using artificial neural network and response surface methodology. J Renew Sustain Energy 5:033104
Torres CA, Marques R, Ferreira AR, Antunes S, Grandfils C, Freitas F, Reis MA (2014) Impact of glycerol and nitrogen concentration on Enterobacter A47 growth and exopolysaccharide production. Int J Biol Macromol 71:81–86
Son MK, Hong SJ, Lee YH (2007) Acetate-mediated pH-stat fed-batch cultivation of transconjugant Enterobacter sp. BL-2S over-expressing glmS gene for excretive production of microbial polyglucosamine PGB-1. J Ind Microbiol Biotechnol 34:799–805
Li L, Li K, Wang Y, Chen C, Xu Y, Zhang L, Han B, Gao C, Tao F, Ma C (2015) Metabolic engineering of Enterobacter cloacae for high-yield production of enantiopure (2R, 3R)-2, 3-butanediol from lignocellulose-derived sugars. Meta Eng 28:19–27
Kleist S, Miksch G, Hitzmann B, Arndt M, Friehs K, Flaschel E (2003) Optimization of the extracellular production of a bacterial phytase with Escherichia coli by using different fed-batch fermentation strategies. Appl Microbiol Biotechnol 61:456–462
Verma D, Satyanarayana T (2012) Phytase production by the unconventional yeast Pichia anomala in fed batch and cyclic fed batch fermentations. Afr J Biotechnol 11:13705–13709
Coban HB, Demirci A (2014) Enhanced submerged Aspergillus ficuum phytase production by implementation of fed-batch fermentation. Bioproc Biosyst Engin 37:2579–2586
Dancer G, Mah JH, Rhee MS, Hwang IG, Kang DH (2009) Resistance of Enterobacter sakazakii (Cronobacter spp.) to environmental stresses. J Appl Microbiol 107:1606–1614
Rowbury R, Goodson M, Wallace A (1992) The PhoE porin and transmission of the chemical stimulus for induction of acid resistance (acid habituation) in Escherichia coli. J Appl Bacteriol 72:233–243
Gauthier MJ, Flatau GN, Clément RL, Munro PM (1993) The loss of culturability by Escherichia coli cells in seawater depends on availability of phosphate ions and phosphate transport systems. Microb Ecol 26:29–35
Zhao W, Xiong A, Fu X, Gao F, Tian Y, Peng R (2010) High level expression of an acid-stable phytase from Citrobacter freundii in Pichia pastoris. Appl Biochem Biotechnol 162:2157–2165
Ghatak A, Guha AK, Ray L (2010) β-d-Galactosidase from Enterobacter cloacae: production and some physicochemical properties. Appl Biochem Biotechnol 162:1678–1688
Cha J, Jung JH, Park SE, Cho MH, Seo DH, Ha SJ et al (2009) Molecular cloning and functional characterization of a sucrose isomerase (isomaltulose synthase) gene from Enterobacter sp. FMB-1. J Appl Microbiol 107:1119–1130
Farrokh P, Yakhchali B, Asghar Karkhane A (2014) Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12. Braz J Microbiol 45:677–687
Graminho ER, Takaya N, Nakamura A, Hoshino T (2015) Purification, biochemical characterization, and genetic cloning of the phytase produced by Burkholderia sp. strain a13. J Gen Appl Microbiol 61:15–23
Yu P, Chen Y (2013) Purification and characterization of a novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsonii ZJ0702. BMC Biotechnol 13:78
Park I, Lee J, Cho J (2012) Degradation of phytate pentamagnesium salt by Bacillus sp. T4 phytase as a potential eco-friendly feed additive. Asian Australas J Anim Sci 10:1466–1472
Yao MZ, Wang X, Wang W, Fu YJ, Liang AH (2013) Improving the thermostability of Escherichia coli phytase, appA, by enhancement of glycosylation. Biotechnol Lett 10:1669–1676
Reddy CS, Achary VM, Manna M, Singh J, Kaul T, Reddy MK (2015) Isolation and molecular characterization of thermostable phytase from Bacillus subtilis (BSPhyARRMK33). Appl Biochem Biotechnol 175:3058–3067
Tang J, Leung A, Leung C, Lim BL (2006) Hydrolysis of precipitated phytate by three distinct families of phytases. Soil Biol Biochem 38:1316–1324
Santos T, Connolly C, Murphy R (2015) Trace element inhibition of phytase activity. Biol Trace Elem Res 163:255–265
Sarıbuga E, Nadaroglu H, Dikbas N, Senol M, Cetin B (2015) Purification, characterization of phytase enzyme from Lactobacillus plantarum bacteria and determination of its kinetic properties. Afr J Biotechnol 13:2373–2378
Bar A, Shinder D, Yosefi S, Vax E, Plavnik I (2003) Metabolism and requirements for calcium and phosphorus in the fast-growing chicken as affected by age. Braz J Nutr 89:51–61
Sugiura SH, Dong FM, Hardy RW (2000) A new approach to estimating the minimum dietary requirement of phosphorus for large rainbow trout based on nonfecal excretions of phosphorus and nitrogen. J Nutr 130:865–872
Tran TT, Hashim SO, Gaber Y, Mamo G, Mattiasson B, Hatti-Kaul R (2011) Thermostable alkaline phytase from Bacillus sp. MD2: effect of divalent metals on activity and stability. J Inorg Biochem 105:1000–1007
Haros M, Rosell CM, Benedito C (2001) Fungal phytase as a potential bread making additive. Eur Food Res Technol 213:317–322
Acknowledgments
The financial support from the National Research Foundation, Republic of South Africa and Durban University of Technology is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they do not have any conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chanderman, A., Puri, A.K., Permaul, K. et al. Production, characteristics and applications of phytase from a rhizosphere isolated Enterobacter sp. ACSS. Bioprocess Biosyst Eng 39, 1577–1587 (2016). https://doi.org/10.1007/s00449-016-1632-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-016-1632-7