The attractive recombinant phytase from Bacillus licheniformis: biochemical and molecular characterization

Springer Science and Business Media LLC - Tập 98 - Trang 5937-5947 - 2013
Mohamed Ali Borgi1, Samira Boudebbouze2, Nushin Aghajari3, Florette Szukala2, Nicolas Pons2, Emmanuelle Maguin2, Moez Rhimi2
1Faculty of Sciences of Gafsa–Unit of Macromolecular Biochemistry and Genetic, Department of Life Sciences, Zarroug, Tunisia
2INRA, UMR 1319 Micalis, Jouy-en-Josas, France
3Laboratory for Biocrystallography and Structural Biology of Therapeutical Targets, BMSSI–Institut de Biologie et Chimie des Protéines, UMR 5086-CNRS/Université de Lyon, Lyon cedex 07, France

Tóm tắt

The phyL gene encoding phytase from the industrial strain Bacillus licheniformis ATCC 14580 (PhyL) was cloned, sequenced, and overexpressed in Escherichia coli. Biochemical characterization demonstrated that the recombinant enzyme has an apparent molecular weight of nearly 42 kDa. Interestingly, this enzyme was optimally active at 70–75 °C and pH 6.5–7.0. This enzyme is distinguishable by the fact that it preserved more than 40 % of its activity at wide range of temperatures from 4 to 85 °C. This new phytase displayed also a high specific activity of 316 U/mg. For its maximal activity and thermostability, this biocatalyst required only 0.6 mM of Ca2+ ion and exhibited high catalytic efficiency of 8.3 s−1 μM−1 towards phytic acid.

Tài liệu tham khảo

Angel R, Tamim NM, Applegate TJ, Dhandu AS, Ellestad LE (2002) Phytic acid chemistry: influence on phytin–phosphorus availability and phytase efficacy. J Appl Poult Res 11:471–480 Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Wiley, New York Boyce A, Walsh G (2006) Comparison of selected physicochemical characteristics of commercial phytases relevant to their application in phosphate pollution abatement. J Environ Sci Health A 41:789–798 Bradford MM (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254 Brugger R, SimoesNunes C, Hug D, Vogel K, Guggenbuhl P, Mascarello F, Augem S, Wyss M, van Loon APGM, Pasamontes L (2004) Characteristics of fungal phytases from Aspergillus fumigatus and Sartorya fumigata. Appl Microbiol Biotechnol 63:383–389 Cao L, Wang W, Yang C, Yang Y, Diana J, Yakupitiyage A, Luo Z, Li D (2007) Application of microbial phytase in fish feed. Enzyme Microb Tech 40:497–507 Cheryan M (1980) Phytic acid interactions in food systems. Crit Rev Food Sci 13:297–335 Cho EA, Kim EJ, Pan JG (2011) Adsorption immobilization of Escherichia coli phytase on probiotic Bacillus polyfermenticus spores. Enzyme Microb Tech 49:66–71 Choi YM, Suh HJ, Kim JM (2001) Purification and properties of extracellular phytase from Bacillus sp. KHU-10. J Protein Chem 20:287–292 Dassa J, Marck C, Boquet PL (1990) The complete nucleotide sequence of the Escherichia coli gene appA reveals significant homology between pH 2.5 acid phosphatase and glucose-1-phosphatase. J Bacteriol 172:5497–5500 Edward JM, Ullah AHJ (2003) The term phytase comprises several different classes of enzymes. Biochem Biophys Res Commun 312:179–184 Escobin-Mopera L, Ohtani M, Sekiguchi S, Sone T, Abe A, Tanaka M, Meevootisom V, Asano K (2012) Purification and characterization of phytase from Klebsiella pneumoniae 9-3B. J of Biosci Bioeng 113:562–567 Farhat A, Chouayekh H, Ben Farhat M, Bouchaala K, Bejar S (2008) Gene cloning and characterization of a thermostable phytase from Bacillus subtilis US417 and assessment of its potential as a feed additive in comparison with a commercial enzyme. Mol Biotechnol 40:127–135 Fu S, Sun J, Qian L (2008a) Bacillus phytases: present scenario and future perspectives. Appl Biochem Biotechnol 151:1–8 Fu S, Sun J, Qian L (2008b) Effect of Ca2+ on beta-propeller phytases. Protein Peptide Lett 15:39–42 Gouet P, Robert X, Courcelle E (2003) ESPript/ENDscript: extracting and rendering sequence and 3D information from atomic structures of proteins. Nucleic Acids Res 31:3320–3323 Greiner R, Konietzny U (2006) Phytase for food application. Food Technol Biotechnol 44:125–140 Gulati HK, Chadha BS, Saini HS (2007) Production and characterization of thermostable alkamine phytase from Bacillus laevolacticus isolated from rhizosphere soil. J Ind Microbiol Biotechnol 34:91–98 Haefner S, Knietsch A, Scholten E, Braun J, Lohscheidt M, Zelder O (2005) Biotechnological production and applications of phytases. Appl Microbiol Biotechnol 68:588–597 Huang H, Luo H, Wang Y, Fu D, Shao N, Yang P, Meng K, Yao B (2009a) Novel low-temperature-active phytase from Erwinia carotovora var. carotovota ACCC 10276. J Microbiol Biotechnol 19:1085–1091 Huang H, Shao N, Wang Y, Luo H, Yang P, Zhou Z, Zhan Z, Yao B (2009b) A novel beta-propeller phytase from Pedobacter nyackensis MJ11 CGMCC 2503 with potential as an aquatic feed additive. Appl Microbiol Biotechnol 83:249–259 In MJ, Seo SW, Oh NS (2008) Fermentative production and application of acid phytase by Saccaromyces cerevisiae CY strain. Afr J Biotechnol 17:3115–3120 Kabsch W, Sander C (1983) Dictionary of protein secondary structure:pattern recognition of hydrogen-bonded and geometrical features. Biopolymers 22:2577–2673 Kerovuo J, Tynkkynen S (2000) Expression of Bacillus subtilis phytase in Lactobacillus plantarum 755. Lett Appl Microbiol 30:325–329 Kerovuo J, Lauraeus M, Nurminen P, Kalkkinen N, Apajalahti J (1998) Isolation, characterization, molecular gene cloning, and sequencing of a novel phytase from Bacillus subtilis. Appl Environ Microbiol 64:2079–2085 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 Lambrechts C, Boze H, Moulin G, Galzy P (1992) Utilization of phytate by some yeast. Biotechnol Lett 14:61–66 Li X, Chi Z, Liu Z, Li J, Wang X (2009) Molecular cloning, characterization, and expression of the phytase gene from marine yeast Kodamaea ohmeri BG3. Mycol Res 113:24–32 Li Z, Zhao A, Wang X, Jin X, Li J, Yu M (2013) Cloning, overexpression, and functional characterization of a phytase from the genus Bacillus. J Mol Microb Biotech 23:193–202 Maenz DD, Engele-Schaan CM, Newkirk RW, Classen HL (1999) The effect of minerals and mineral chelators on the formation of phytase-resistant and phytase-susceptible forms of phytic acid in solution and in a slurry of canola meal. Animal Feed Scien Technol 81:177–192 Martinez AC, Parsons CM, Baker DH (2006) Effect of microbial phytase and citric acid on phosphorus bioavailability, apparent metabolizable energy, and amino acid digestibility in distillers dried grains with solubles in chicks. Poultry Sci 85:470–475 Mayer AF, Hellmuth K, Schlieker H, Lopez-Ulibarri R, Oertel S, Dahlems U, Strasser AW, van Loon AP (1999) An expression system matures: a highly efficient and cost-effective process for phytase production by recombinant strains of Hansenula polymorpha. Biotechnol Bioeng 63:373–381 Mullaney EJ, Daly CB, Ullah AHJ (2000) Advances in phytase research. Adv Appl Microbiol 47:157–199 Nayini NR, Markakis P (1984) The phytase of yeast. Food SciTechnol 17:126–132 Ni Y, Chen R (2009) Extracellular recombinant protein production from Escherichia coli. Biotechnol Lett 31:1661–1670 Oh BC, Chang BS, Park KW, Ha NC, Kim HK, Oh BH, Oh TK (2001) Calcium-dependent catalytic activity of a novel phytase from Bacillus amyloliquefaciens DS11. Biochemistry 40:9669–9676 Oh BC, Choi WC, Park S, Kim YO, Oh TK (2004) Biochemical properties and substrate specificities of alkaline and histidine acid phytases. Appl Microbiol Biotechnol 63:362–372 Ostanin K, Harms EH, Stevis PE, Kuciel R, Zhou MM, van Etten RL (1992) Overexpression, site-directed mutagenesis, and mechanism of Escherichia coli acid phosphatase. J Biol Chem 267:22830–22836 Park SC, Choi YW, Oh TK (1999) Comparative enzymatic hydrolysis of phytate in various animal feedstuff with two different phytases. J Vet Med Sci 61:1257–1259 Parry R (1998) Agricultural phosphorus and water quality: a U.S. Environmental Protection Agency perspective. J Environ Qual 27:258–261 Ragon M, Neugnot-Roux V, Chemardin P, Moulin G, Boze H (2008) Molecular gene cloning and overexpression of the phytase from Debaryomyces castellii CBS 2923. Protein Expres Purif 58:275–283 Reddy NR, Sathe SK, Salunkhe DK (1982) Phytates in legumes and cereals. Adv Food Res 28:1–92 Rey MW, Ramaiya P, Nelson BA, Brody-Karpin SD, Zaretsky EJ, Tang M, Lopez de Leon A, Xiang H, Gusti V, Clausen IG, Olsen PB, Rasmussen MD, Andersen JT, Jørgensen PL, Larsen TS, Sorokin A, Bolotin A, Lapidus A, Galleron N, Ehrlich SD, Berka RM (2004) Complete genome sequence of the industrial bacterium Bacillus licheniformis and comparisons with closely related Bacillus species. Genome Biol 10:R77 Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor Selle PH, Ravindran V (2007) Microbial phytase in poultry nutrition. Anim Feed Sci Tech 135:1–41 Sharpley A, Daniel TC, Sims JT, Pote DH (1996) Determining environmentally sound soil phosphorus levels. J Soil Water Conserv 51:160–166 Shi P, Huang H, Wang Y, Luo H, Wu B, Meng K, Yang P, Yao B (2008) A novel phytase gene appA from Buttiauxella sp. GC21 isolated from grass carp intestine. Aquaculture 275:70–75 Shin S, Ha NC, Oh BC, Oh TK, Oh BH (2001) Enzyme mechanism and catalytic property of beta propeller phytase. Structure 9:851–858 Shivange AV, Serwe A, Dennig A, Roccatano D, Haefner S, Schwaneberg U (2012) Directed evolution of a highly active Yersinia mollaretii phytase. Appl Microbiol Biotechnol 95:405–418 Sohail SS, Roland DA (1999) Influence of supplemental phytase on performance of broilers four to six weeks of age. Poult Sci 78:550–555 Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739 Thompson JD, Higgins DG, Gibson TJ, (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 Tomaschy A, Brugger R, Lehmann M, Svendsen A, Vogel K, Kostrewa D, Lassen SF, Burger D, Kronenberger A, van Loon APGM, Pasamontes L, Wyss M (2002) Engineering of phytase for improved activity at low pH. Appl Environ Microb 68:1907–1913 Tran TT, Mamo G, Mattiasson B, Hatt-Kaul R (2010) A thermostable phytase from Bacillus sp. MD2: cloning, expression and high-level production in Escherichia coli. J Ind Microbiol Biotechnol 37:279–287 Tran TT, Mamo G, Buxo L, Le NN, Gaber Y, Mattiasson B, Hatt-Kaul R (2011) Site-directed mutagenesis of an alkaline phytase: influencing specificity, activity and stability in acidic milieu. Enzyme Microb Tech 49:177–182 Tye AJ, Siu FKY, Leung TYC, Lim BL (2002) Molecular cloning and the biochemical characterization of two novel phytases from B. subtilis 168 and B. licheniformis. Appl Microbiol Biotechnol 59:190–197 Ullah AH, Gibson DM (1987) Extracellular phytase (E.C. 3.1.3.8) from Aspergillus ficuum NRRL 3135: purification and characterization. Prep Biochem 17:63–91 Vats P, Banerjee UC (2004) Production studies and catalytic properties of phytases (myo-inositol hexakisphosphate phosphohydrolases): an overview. Enzyme Microb Tech 35:3–14 Watanabe T, Ikeda H, Masaki K, Fujii T, Iefuji H (2009) Cloning and characterization of a novel phytase from wastewater treatment yeast Hansenula fabianii J640 and expression in Pichia pastoris. J Biosci Bioeng 108:225–230 Yao MZ, Zhang YH, Lu WL, Hu MQ, Wang W, Liang AH (2011) Phytases: crystal structures, protein engineering and potential biotechnological applications. J Appl Microbiol 112:1–14 Zang GQ, Dong XF, Wang ZH, Zhang Q, Wang HX Tong JM (2010) Purification, characterization, and cloning of a novel phytase with low pH optimum and strong proteolysis resistance from Aspergillus ficuum NTG-23. BioresourceTechnol 101:4125–4131 Zeng YF, Ko TP, Lai HL, Cheng YS, Wu TH, Ma Y, Chen CC, Yang CS, Cheng KJ, Huang CS, Guo RT, Liu JR (2011) Crystal structure of Bacillus alkaline phytase in complex with divalent metal ions and inositol hexasulfate. J Mol Biol 409:214–224 Zhang W, Lei XG (2008) Cumulative improvements of thermostability and pH-activity profile of Aspergillus niger PhyA phytase by site-directed mutagenesis. Appl Microbiol Biotechnol 77:1033–1040