Abstract
β-propeller phytase-like sequences (BPP-like sequences) are widespread in the microbial world and have been found in the sequenced genomes of aquatic, soil, and plant bacteria. Exploring NCBI microbial genome database for putative genes encoding phytase, a BPP-like sequence from Sphingomonas wittichii RW-1 (Sequence ID: CP000699.1), known for its capacity of degrading polychlorinated dibenzo-p-dioxins and dibenzofurans, was recognized. The putative phytase gene (phySw) was amplified with specific primers, cloned, and overexpressed in Escherichia coli and the catalytic properties of the recombinant PhySw protein were analyzed. The results show that phySw encodes an enzyme with the properties of β-propeller phytases: it requires the presence of Ca2+ ions, it is optimally active at 55 °C, and it has a pH optimum of 6.0 with good activity in the range 6.0–8.0. Furthermore, the enzyme exhibits a good thermostability, recovering 68% of its original activity after treatment at 80 °C for 10 min, and shows a good substrate specificity for phytic acid. These properties render this enzyme a candidate as an animal feed additive (e.g., for aquaculture industry). The isolation of phytases from a hydrocarbon-utilizing microorganism also opens new scenarios for their possible application in combating oil pollution.
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References
Aislabie J, Saul DJ, Foght JM (2006) Bioremediation of hydrocarbon-contaminated polar soils. Extremophiles 10:171–179. https://doi.org/10.1007/s00792-005-0498-4
Bae HD, Yanke LJ, Cheng KJ, Selinger LB (1999) A novel staining method for detecting phytase activity. J Microbiol Methods 39:17–22
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
Brinch-Pedersen H, Sorensen LD, Holm PB (2002) Engineering crop plants: getting a handle on phosphate. Trends Plant Sci 7:118–125
Chai B, Tsoi TV, Iwai S, Liu C, Fish JA, Gu C, Johnson TA, Zylstra G, Teppen BJ, Li H, Hashsham SA, Boyd SA, Cole JR, Tiedje JM (2016) Sphingomonas wittichii strain RW-1 genome-wide gene expression shifts in response to dioxins and clay. PLoS One 11:1–14. https://doi.org/10.1371/journal.pone.0157008
Cheng C, Lim BL (2006) Beta-propeller phytases in the aquatic environment. Arch Microbiol 185:1–13. https://doi.org/10.1007/s00203-005-0080-6
Cheng C, Wong K-B, Lim BL (2007) The effect of disulfide bond on the conformational stability and catalytic activity of beta-propeller phytase. Protein Pept Lett 14:175–183
Dersjant-Li Y, Awati A, Schulze H, Partridge G (2015) Phytase in non-ruminant animal nutrition: a critical review on phytase activities in the gastrointestinal tract and influencing factors. J Sci Food Agric 95:878–896. https://doi.org/10.1002/jsfa.6998
Ferguson AD, Deisenhofer J (2004) Metal import through microbial membranes. Cell 116:15–24. https://doi.org/10.1016/S0092-8674(03)01030-4
Garrett JB, Kretz KA, O’Donoghue E, Kerovuo J, Kim W, Barton NR, Hazlewood GP, Short JM, Robertson DE, Gray KA (2004) Enhancing the thermal tolerance and gastric performance of a microbial phytase for use as a phosphate-mobilizing monogastric-feed supplement. Appl Environ Microbiol 70:3041–3046. https://doi.org/10.1128/AEM.70.5.3041-3046.2004
Gerke J (2015) Phytate (inositol hexakisphosphate) in soil and phosphate acquisition from inositol phosphates by higher plants. A Review Plants 4:253–266. https://doi.org/10.3390/plants4020253
Ha NC, Oh BC, Shin S, Kim HJ, Oh TK, Kim YO, Choi KY, Oh BH (2000) Crystal structures of a novel, thermostable phytase in partially and fully calcium-loaded states. Nat Struct Biol 7:147–153. https://doi.org/10.1038/72421
Han Y, Wilson DB, Lei X (1999) Expression of an Aspergillus niger phytase gene ( phyA) in Saccharomyces cerevisiae. Appl Environ Microbiol 65:1915–1918
Huang H, Shi P, Wang Y, Luo H, Shao N, Wang G, Yang P, Yao B (2009) Diversity of beta-propeller phytase genes in the intestinal contents of grass carp provides insight into the release of major phosphorus from phytate in nature. Appl Environ Microbiol 75:1508–1516. https://doi.org/10.1128/AEM.02188-08
Jorquera MA, Gabler S, Inostroza NG, Acuña JJ, Campos MA, Menezes-Blackburn D, Greiner R (2018) Screening and characterization of phytases from bacteria isolated from Chilean hydrothermal environments. Microb Ecol 75:387–399. https://doi.org/10.1007/s00248-017-1057-0
Kim TW, Lei XG (2005) An improved method for a rapid determination of phytase activity in animal feed. J Anim Sci 83:1062–1067
Kumar V, Sinha AK, Makkar HPS, Becker K (2010) Dietary roles of phytate and phytase in human nutrition: a review. Food Chem 120:945–959. https://doi.org/10.1016/j.foodchem.2009.11.052
Kumar V, Yadav AN, Verma P, Sangwan P, Saxena A, Kumar K, Singh B (2017) β-propeller phytases: diversity, catalytic attributes, current developments and potential biotechnological applications. Int J Biol Macromol 98:595–609. https://doi.org/10.1016/j.ijbiomac.2017.01.134
Lim BL, Yeung P, Cheng C, Hill JE (2007) Distribution and diversity of phytate-mineralizing bacteria. ISME J 1:321–330. https://doi.org/10.1038/ismej.2007.40
Mallin MA, Cahoon LB (2003) Industrialized animal production—a major source of nutrient and microbial pollution to aquatic ecosystems. Popul Environ 24:369–386. https://doi.org/10.1023/A:1023690824045
Martin CJ, Evans WJ (1986) Phytic acid-metal ion interactions. II. The effect of pH on Ca(II) binding. J Inorg Biochem 27:17–30. https://doi.org/10.1016/0162-0134(86)80105-2
Miller TR, Delcher AL, Salzberg SL, Saunders E, Detter JC, Halden RU (2010) Genome sequence of the dioxin-mineralizing bacterium Sphingomonas wittichii RW1. J Bacteriol 192:6101–6102. https://doi.org/10.1128/JB.01030-10
Muslim SN, Mohammed Ali AN, AL-Kadmy IMS, Khazaal SS, Ibrahim SA, Al-Saryi NA, Al-Saadi LG, Muslim SN, Salman BK, Aziz SN (2018) Screening, nutritional optimization and purification for phytase produced by Enterobacter aerogenes and its role in enhancement of hydrocarbons degradation and biofilm inhibition. Microb Pathog 115:159–167. https://doi.org/10.1016/j.micpath.2017.12.047
Nakai K, Horton P (1999) PSORT: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci 24:34–36
Oh BC, Chang BS, Park KH, 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. https://doi.org/10.1007/s00253-003-1345-0
Raboy V (1997) Accumulation and storage of phosphate and minerals. In: Larkins BA, Vasil IK (eds) Cellular and molecular biology of plant seed development. Springer Netherlands, Dordrecht, pp 441–477
Ranjan B, Satyanarayana T (2016) Recombinant HAP Phytase of the thermophilic mold Sporotrichum thermophile: expression of the codon-optimized phytase gene in Pichia pastoris and applications. Mol Biotechnol 58:137–147. https://doi.org/10.1007/s12033-015-9909-7
Santos T, Connolly C, Murphy R (2015) Trace element inhibition of phytase activity. Biol Trace Elem Res 163:255–265. https://doi.org/10.1007/s12011-014-0161-y
Scholey D, Burton E, Morgan N, Sanni C, Madsen CK, Dionisio G, Brinch-Pedersen H (2017) P and Ca digestibility is increased in broiler diets supplemented with the high-phytase HIGHPHY wheat. Animal 11:1457–1463. https://doi.org/10.1017/S1751731117000544
Sebastian S, P. Touchburn S, Chavez RE (1998) Implications of phytic acid and supplemental microbial phytase in poultry nutrition: a review. Worlds Poultry Sci 54:27–47. https://doi.org/10.1079/WPS19980003
Seshadri R, Kravitz SA, Smarr L, Gilna P, Frazier M (2007) CAMERA: a community resource for metagenomics. PLoS Biol 5:0394–0397. https://doi.org/10.1371/journal.pbio.0050075
Ushasree MV, Shyam K, Vidya J, Pandey A (2017) Microbial phytase: impact of advances in genetic engineering in revolutionizing its properties and applications. Bioresour Technol 245:1790–1799. https://doi.org/10.1016/j.biortech.2017.05.060
Wealleans AL, Bold RM, Dersjant-Li Y, Awati A (2015) The addition of a Buttiauxella sp. phytase to lactating sow diets deficient in phosphorus and calcium reduces weight loss and improves nutrient digestibility. J Anim Sci 93:5283–5290. https://doi.org/10.2527/jas.2015-9317
Wyss M, Brugger R, Kronenberger A, Rémy R, Fimbel R, Oesterhelt G, Van Loon APGM, Re R, Lehmann M (1999) Biochemical characterization of fungal phytases ( myo-inositol hexakisphosphate phosphohydrolases ): catalytic properties biochemical characterization of fungal Phytases ( myo -inositol hexakisphosphate Phosphohydrolases ): catalytic properties. Appl Environ Microbiol 65:367–373
Yabuuchi E, Yamamoto H, Terakubo S, Okamura N, Naka T, Fujiwara N, Kobayashi K, Kosako Y, Hiraishi A (2001) Proposal of Sphingomonas wittichii sp. nov. for strain RW1T, known as a dibenzo-p-dioxin metabolizer. Int J Syst Evol Microbiol 51:281–292. https://doi.org/10.1099/00207713-51-2-281
Yao MZ, Lu WL, Chen TG, Wang W, Fu YJ, Yang BS, Liang AH (2014) Effect of metals ions on thermostable alkaline phytase from Bacillus subtilis YCJS isolated from soybean rhizosphere soil. Ann Microbiol 64:1123–1131. https://doi.org/10.1007/s13213-013-0751-5
Zeng ZK, Li QY, Zhao PF, Xu X, Tian QY, Wang HL, Pan L, Yu S, Piao XS (2016) A new Buttiauxella phytase continuously hydrolyzes phytate and improves amino acid digestibility and mineral balance in growing pigs fed phosphorous-deficient diet. J Anim Sci 94:629–638. https://doi.org/10.2527/jas2015-9143
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The authors are also grateful to Technopole SITEIA-Parma (Regione Emilia-Romagna) and Prof Nelson Marmiroli for allowing access to technical infrastructures.
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This research was in part supported by funding to Prof. Anna Maria Sanangelantoni from FIL of the University of Parma (Local Funding for Research).
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Sanangelantoni, A.M., Malatrasi, M., Trivelloni, E. et al. A novel β-propeller phytase from the dioxin-degrading bacterium Sphingomonas wittichii RW-1. Appl Microbiol Biotechnol 102, 8351–8358 (2018). https://doi.org/10.1007/s00253-018-9248-2
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DOI: https://doi.org/10.1007/s00253-018-9248-2