Effect of purified alkaline phosphatase from Bacillus licheniformis on growth of Zea mays L.
DOI:
https://doi.org/10.14719/pst.2019.6.sp1.676Keywords:
Alkaline phosphatase, Bacillus licheniformis, Biofertilizer, Zea maysAbstract
Some soil microbes have the capability to solubilize mineral phosphate into organic phosphorous and used as biofertilizer to improve crop productivity in agricultural field. In this study, phosphate solubilization assay was carried out onto media plates containing calcium phsophate precipitated nutrient agar media for bacterial strains like Bacillus megaterium MTCC 453, Bacillus subtilis MTCC 1134, Bacillus licheniformis MTCC 2312, Pseudomonas aeruginosa MTCC 424, Escherichia coli MTCC 570. Among these bacterial strains, B. licheniformis MTCC 2312 showed largest clear zone of phosphate solubilzation and maximum activity of alkaline phosphatase. The enzyme alkaline phosphatase was purified from B. licheniformis MTCC 2312 with purification fold 3.52 and specific activity 295.89 Unit/mg protein using DEAE-sepharose chromatography. This enzyme showed molecular weight as 60 KD, thermostability upto 50?C, pH stability up to 8.5 and Michaelis constant (Km) and maximum activity (Vmax) as 2.30 mM and 2223 U/ml respectively. The lyophilized powder of this enzyme was further supplemented with media components for the growth of Zea mays for carrying tissue culture experiment. The sterilized soil supplemented with alkaline phosphatase improved the total height, dry weight, % phosphate content in the stem and root of Zea mays by 3.07, 3.15, 2.35 and 1.76 fold respectively compared to control set. This enzyme could be used at large extent as effective biofertilizer for the agricultural industry.
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References
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9. Jahan M, Mahallati MN, Amiri MB, Ehyayi HR. Radiation absorption and use efficiency of sesame as affected by biofertilizers inoculation, in a low input cropping system. Industrial Crops and products. 2013;43:606–11. https://doi.org/10.1016/j.indcrop.2012.08.012
10. David P, Raj RS, Linda R, Rhema SB. Molecular characterization of phosphate solubilizing bacteria (PSB) and plant growth promoting rhizobacteria (PGPR) from pristine soils. International Journal of Innovative Science Engineering and Technology. 2014;1:317–24.
11. Mamta RP, Pathania V, Gulati A, Singh B, Bhanwra RK, Tewari R. Stimulatory effect of phosphate-solubilizing bacteria on plant growth, stevioside and rebaudioside-A contents of Stevia rebaudiana Bertoni. Applied Soil Ecology. 2010;46:222–29. https://doi.org/10.1016/j.apsoil.2010.08.008
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13. Istina IN, Widiastuti H, Joy B, Antralina M. Phosphate solubilizing microbe from Saprists peat soil and their potency to enhance oil palm growth and P uptake. Procidia Food Science. 2015;3:426–35. https://doi.org/10.1016/j.profoo.2015.01.047
14. Chakraborty U, Chakraborty BN, Basnet M, Chakraborty, AP. Evaluation of Ochrobactrum anthropi TRS-2 and its talc based formulation for enhancement of growth of tea plants and management of brown root rot disease. Journal of Applied Microbiology. 2009;107:625–34. https://doi.org/10.1111/j.1365-2672.2009.04242.x
15. Fernandez Bidondo L, Silvani V, Colombo R, Pergola M, Bompadre J, Godeas A. Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host. Soil Biology and Biochemistry. 2011;43:1866–72. https://doi.org/10.1016/j.soilbio.2011.05.004
16. Halder AK, Mishra AK, Bhattacharya P, Chakrabarty PK. Solubilization of rock phosphate by Rhizobium and Bradyrhizobium. Journal of General Applied Microbiology. 1990;36:81–92.
17. Alori ET, Glick BR, Babalola OO. Microbial phosphorus solubilization and its potential for use in sustainable agriculture. Frontiers in Microbiology. 2017;8:971. https://doi.org/10.3389/fmicb.2017.00971
18. Surange S, Wollum II AG, Kumar N, Nautiyal CS. Characterization of Rhizobium from root nodules of leguminous trees growing in alkaline soils. Canadian Journal of Microbiology. 1997;43:891–94. https://doi.org/10.1139/m97-130
19. Gaind S, Gaur AC. Thermotolerant phosphate solubilizing microorganisms and their interaction with mung bean. Plant Soil 1991;133:141–49. https://doi.org/10.1007/BF00011908
20. Liu M, Liu X, Cheng BS, Ma XL, Lyu XT, Zhao XF, et al. Selection and evaluation of phosphate-solubilizing bacteria from grapevine rhizospheres for use as biofertilizers. Spanish Journal of Agricultural Research. 2016;14:4. https://doi.org/10.5424/sjar/2016144-9714
21. El-Sersy NA, Ebrahim HAH, Abou-Elela GM. Response surface methodology as a tool for optimizing the production of antimicrobial agents from Bacillus licheniformis SN2. Current Research in Bacteriology. 2010;3(1):1-14. https://doi.org/10.3923/crb.2010.1.14
22. Garen A, Levinthal C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. 1 - Purification and characterization of alkaline phosphatase. Biochimica et Biophysica Acta. 1960;38:470-83. https://doi.org/10.1016/0006-3002(60)91282-8
23. Lowry OH, Lopez JA. The determination of inorganic phosphate in the presence of labile phosphate esters. Journal of Biological Chemistry. 1946;162:421-28.
24. Darmwall NS, Singh RB, Rai R. Isolation of phosphate solubilizers from different sources. Current Science. 1989;58:570–71.
25. Bardiya MC, Gaur AC. Isolation and screening of microorganisms dissolving low grade rock phosphate. Folia Microbiology. 1974;19:386–89. https://doi.org/10.1007/BF02872824
26. Katznelson H, Peterson EA, Rovatt JW. Phosphate dissolving microoganisms on seed and in the root zone of plants. Canadian Journal of Botany. 1962;40:1181–86. https://doi.org/10.1139/b62-108
27. Kostadinova S, Marhova M. Purification and Properties of Alkaline Phosphatase from Bacillus cereus. Biotechnology & Biotechnological Equipment. 2010;24:602-06. https://doi.org/10.1139/b62-108
28. Dhaked RK, Alam SI, Dixit A, Singh L. Purification and characterization of thermolabile alkaline phosphatase from an Antarctic psychrotolerant Bacillus sp. P9. Enzyme Microbial Technology. 2005;36:855–61. https://doi.org/10.1016/j.enzmictec.2004.11.017
29. Goldman S, Hecht K, Eisenberg H, Mevarech M. Extracellular Ca2+-dependent inducible alkaline phosphatase from the extremely halophilic archeabacterium Haloarcula marismortui. Journal of Bacteriology. 1990; 172:7065–70. https://doi.org/10.1128/JB.172.12.7065-7070.1990
30. Fitt PS, Peterkin PI. Isolation and properties if a small manganese-ion- stimulated bacterial alkaline phosphatase. Biochemical Journal. 1976;157:161–67. https://doi.org/10.1042/bj1570161
31. Posen S. Alkaline phosphatase. Annals of Internal Medicine. 1967;67:183–203. https://doi.org/10.7326/0003-4819-67-1-183
32. Morales AC, Nozawa SR, Thedei G, Maccheroni W, Rossi A. Properties of a constitutive alkaline phosphatase from strain 74A of the mold Neurospora crassa. Brazilian Journal of Medical and Biological Research. 2000;33:905–12. https://doi.org/10.1590/S0100-879X2000000800006
33. Yeh MF, Trela JM. Purification and characterization of a repressible alkaline phosphatase from Thermus aquaticus. Journal of Biological Chemistry. 1976;251:3134-39.
34. Dong GQ, Zeikus JG. Purification and characterization of alkaline phosphatase from Thermotoga neapolitana. Enzyme Microbial Technology. 1997;21:335–40. https://doi.org/10.1016/S0141-0229(97)00002-1
35. Wojciechowski CL, Cardia JP, Kantrowitz ER. Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity. Protein Science. 2002;11:903–11. https://doi.org/10.1110/ps.4260102
36. Duff RB, Webley DM. 2-Ketogluconic acid as a natural chelator produced by soil bacteria. Chemistry and Industry (London). 1959;1376–77.
37. Banik S, Dey BK. Available phosphate content of an alluvial soil is influenced by inoculation of some isolated phosphate-solubilizing microorganisms. Plant Soil. 1982;69:353–64. https://doi.org/10.1007/BF02372456
38. Ohtake H, Wu H, Imazu K, Ambe Y, Kato J, Kuroda A. Bacterial phosphonate degradation, phosphite oxidation and polyphosphate accumulation. Resource Conservation and Recycling. 1996;18:125–34. https://doi.org/10.1016/S0921-3449(96)01173-1
39. McGrath JW, Wisdom GB, McMullan G, Lrakin MJ, Quinn, JP. The purification and properties of phosphonoacetate hydrolase, a novel carbon-phosphorus bond-cleaving enzyme from Pseudomonas fluorescens 23F. European Journal of Biochemistry. 1995;234:225–30. https://doi.org/10.1111/j.1432-1033.1995.225_c.x
40. Bujacz B, Wieczorek P, Krzysko-Lupcka T, Golab Z, Lejczak B, Kavfarski P. Organophosphonate utilization by the wild-type strain of Penicillium notatum. Applied Environmental Microbiology. 1995;61:2905–10. https://doi.org/10.1128/AEM.61.8.2905-2910.1995
41. Krasilnikov M. On the role of soil bacteria in plant nutrition. Journal of General and Applied Microbiology. 1961;7:128–44. https://doi.org/10.2323/jgam.7.128
42. Hall JA, Pierson D, Ghosh S, Glick BR. Root elongation in various agronomic crops by the plant growth promoting rhizobacterium Pseudomonas putida GR12-2. Israel Journal of Plant Sciences. 1996;44:37–42. https://doi.org/10.1080/07929978.1996.10676631
43. Glick BR, Changping L, Sibdas G, Dumbroff EB. Early development of canola seedlings in the presence of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2. Soil Biological Biochemistry. 1997;29:1233–39. https://doi.org/10.1016/S0038-0717(97)00026-6
44. Kloepper JW, Lifshitz K, Schroth MN. Pseudomonas inoculants to benefit plant production. ISI Atlas of Science: Animal and Plant Sciences. 1988;60–64.
45. Kapulnik J, Gafny R, Okon Y. Effect of Azopirillum spp. inoculation on root development and NO-3 uptake in wheat (Titicum aestivum cv. Miriam) in hydroponic systems. Canadian Journal of Botany. 1985;63:627–31. https://doi.org/10.1139/b85-078
46. Broadbent P, Baker KF, Franks N, Holland J. Effect of Bacillus spp. on increased growth of seedlings in steamed and in nontreated soil. Phytopathology. 1977;67:1027–34. https://doi.org/10.1094/Phyto-67-1027
47. Burr TJ, Schroth MN, Suslow T. Increased potato yields by treatment of seedpieces with specific strains of Pseudomonas fluorescens and Pseudomonas putida. Phytopathology. 1978;68:1377–83. https://doi.org/10.1094/Phyto-68-1377
2. Sharma SB, Sayyed RZ, Trivedi MH, Gobi TA. Phosphate solubilizing microbes: sustainable approach for managing phosphorus deficiency in agricultural soils. Springer plus. 2013;2:587–600. https://doi.org/10.1186/2193-1801-2-587
3. Kalayu G. Phosphate solubilizing microorganisms: promising approach as biofertilizers. International Journal of Agronomy. 2019;1-7.
4. Halvorson HO, Keynan A, Kornberg HL. Utilisation of calcium phosphates for microbial growth at alkaline pH. Soil Biological Biochemistry. 1990;22:887–90.
5. Azziz G, Bajsa N, Haghjou T, Taule C, Valverde A, Igual JM, Arias A. Abundance, diversity and prospecting of culturable phosphate solubilizing bacteria on soils under crop–pasture rotations in a no-tillage regime in Uruguay. Applied Soil Ecology. 2012;61:320–26. https://doi.org/10.1016/j.apsoil.2011.10.004
6. Tak HI, Ahmad F, Babalola OO, Inam A. Growth, photosynthesis and yield of chickpea as influenced by urban wastewater and different levels of phosphorus. International Journal of Plant Research. 2012;2:6–13. https://doi.org/10.5923/j.plant.20120202.02
7. Babalola OO, Glick BR. The use of microbial inoculants in African agriculture: current practice and future prospects. Journal of Food, Agriculture, and Environment. 2012b; 540–49.
8. Kumar S, Bauddh K, Barman SC, Singh, RP. Amendments of microbial bio fertilizers and organic substances reduces requirement of urea and DAP with enhanced nutrient availability and productivity of wheat (Triticum aestivum L.). Ecological Engineering Journal. 2014;71:432–37. https://doi.org/10.1016/j.ecoleng.2014.07.007
9. Jahan M, Mahallati MN, Amiri MB, Ehyayi HR. Radiation absorption and use efficiency of sesame as affected by biofertilizers inoculation, in a low input cropping system. Industrial Crops and products. 2013;43:606–11. https://doi.org/10.1016/j.indcrop.2012.08.012
10. David P, Raj RS, Linda R, Rhema SB. Molecular characterization of phosphate solubilizing bacteria (PSB) and plant growth promoting rhizobacteria (PGPR) from pristine soils. International Journal of Innovative Science Engineering and Technology. 2014;1:317–24.
11. Mamta RP, Pathania V, Gulati A, Singh B, Bhanwra RK, Tewari R. Stimulatory effect of phosphate-solubilizing bacteria on plant growth, stevioside and rebaudioside-A contents of Stevia rebaudiana Bertoni. Applied Soil Ecology. 2010;46:222–29. https://doi.org/10.1016/j.apsoil.2010.08.008
12. Zhao K, Penttinen P, Zhang X, Ao X, Liu M, Yu X, Chen Q. Maize rhizosphere in Sichuan, China, hosts plant growth promoting Burkholderia cepacia with phosphate solubilizing and antifungal abilities. Microbiological Research. 2014;169:76–82. https://doi.org/10.1016/j.micres.2013.07.003
13. Istina IN, Widiastuti H, Joy B, Antralina M. Phosphate solubilizing microbe from Saprists peat soil and their potency to enhance oil palm growth and P uptake. Procidia Food Science. 2015;3:426–35. https://doi.org/10.1016/j.profoo.2015.01.047
14. Chakraborty U, Chakraborty BN, Basnet M, Chakraborty, AP. Evaluation of Ochrobactrum anthropi TRS-2 and its talc based formulation for enhancement of growth of tea plants and management of brown root rot disease. Journal of Applied Microbiology. 2009;107:625–34. https://doi.org/10.1111/j.1365-2672.2009.04242.x
15. Fernandez Bidondo L, Silvani V, Colombo R, Pergola M, Bompadre J, Godeas A. Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host. Soil Biology and Biochemistry. 2011;43:1866–72. https://doi.org/10.1016/j.soilbio.2011.05.004
16. Halder AK, Mishra AK, Bhattacharya P, Chakrabarty PK. Solubilization of rock phosphate by Rhizobium and Bradyrhizobium. Journal of General Applied Microbiology. 1990;36:81–92.
17. Alori ET, Glick BR, Babalola OO. Microbial phosphorus solubilization and its potential for use in sustainable agriculture. Frontiers in Microbiology. 2017;8:971. https://doi.org/10.3389/fmicb.2017.00971
18. Surange S, Wollum II AG, Kumar N, Nautiyal CS. Characterization of Rhizobium from root nodules of leguminous trees growing in alkaline soils. Canadian Journal of Microbiology. 1997;43:891–94. https://doi.org/10.1139/m97-130
19. Gaind S, Gaur AC. Thermotolerant phosphate solubilizing microorganisms and their interaction with mung bean. Plant Soil 1991;133:141–49. https://doi.org/10.1007/BF00011908
20. Liu M, Liu X, Cheng BS, Ma XL, Lyu XT, Zhao XF, et al. Selection and evaluation of phosphate-solubilizing bacteria from grapevine rhizospheres for use as biofertilizers. Spanish Journal of Agricultural Research. 2016;14:4. https://doi.org/10.5424/sjar/2016144-9714
21. El-Sersy NA, Ebrahim HAH, Abou-Elela GM. Response surface methodology as a tool for optimizing the production of antimicrobial agents from Bacillus licheniformis SN2. Current Research in Bacteriology. 2010;3(1):1-14. https://doi.org/10.3923/crb.2010.1.14
22. Garen A, Levinthal C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. 1 - Purification and characterization of alkaline phosphatase. Biochimica et Biophysica Acta. 1960;38:470-83. https://doi.org/10.1016/0006-3002(60)91282-8
23. Lowry OH, Lopez JA. The determination of inorganic phosphate in the presence of labile phosphate esters. Journal of Biological Chemistry. 1946;162:421-28.
24. Darmwall NS, Singh RB, Rai R. Isolation of phosphate solubilizers from different sources. Current Science. 1989;58:570–71.
25. Bardiya MC, Gaur AC. Isolation and screening of microorganisms dissolving low grade rock phosphate. Folia Microbiology. 1974;19:386–89. https://doi.org/10.1007/BF02872824
26. Katznelson H, Peterson EA, Rovatt JW. Phosphate dissolving microoganisms on seed and in the root zone of plants. Canadian Journal of Botany. 1962;40:1181–86. https://doi.org/10.1139/b62-108
27. Kostadinova S, Marhova M. Purification and Properties of Alkaline Phosphatase from Bacillus cereus. Biotechnology & Biotechnological Equipment. 2010;24:602-06. https://doi.org/10.1139/b62-108
28. Dhaked RK, Alam SI, Dixit A, Singh L. Purification and characterization of thermolabile alkaline phosphatase from an Antarctic psychrotolerant Bacillus sp. P9. Enzyme Microbial Technology. 2005;36:855–61. https://doi.org/10.1016/j.enzmictec.2004.11.017
29. Goldman S, Hecht K, Eisenberg H, Mevarech M. Extracellular Ca2+-dependent inducible alkaline phosphatase from the extremely halophilic archeabacterium Haloarcula marismortui. Journal of Bacteriology. 1990; 172:7065–70. https://doi.org/10.1128/JB.172.12.7065-7070.1990
30. Fitt PS, Peterkin PI. Isolation and properties if a small manganese-ion- stimulated bacterial alkaline phosphatase. Biochemical Journal. 1976;157:161–67. https://doi.org/10.1042/bj1570161
31. Posen S. Alkaline phosphatase. Annals of Internal Medicine. 1967;67:183–203. https://doi.org/10.7326/0003-4819-67-1-183
32. Morales AC, Nozawa SR, Thedei G, Maccheroni W, Rossi A. Properties of a constitutive alkaline phosphatase from strain 74A of the mold Neurospora crassa. Brazilian Journal of Medical and Biological Research. 2000;33:905–12. https://doi.org/10.1590/S0100-879X2000000800006
33. Yeh MF, Trela JM. Purification and characterization of a repressible alkaline phosphatase from Thermus aquaticus. Journal of Biological Chemistry. 1976;251:3134-39.
34. Dong GQ, Zeikus JG. Purification and characterization of alkaline phosphatase from Thermotoga neapolitana. Enzyme Microbial Technology. 1997;21:335–40. https://doi.org/10.1016/S0141-0229(97)00002-1
35. Wojciechowski CL, Cardia JP, Kantrowitz ER. Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity. Protein Science. 2002;11:903–11. https://doi.org/10.1110/ps.4260102
36. Duff RB, Webley DM. 2-Ketogluconic acid as a natural chelator produced by soil bacteria. Chemistry and Industry (London). 1959;1376–77.
37. Banik S, Dey BK. Available phosphate content of an alluvial soil is influenced by inoculation of some isolated phosphate-solubilizing microorganisms. Plant Soil. 1982;69:353–64. https://doi.org/10.1007/BF02372456
38. Ohtake H, Wu H, Imazu K, Ambe Y, Kato J, Kuroda A. Bacterial phosphonate degradation, phosphite oxidation and polyphosphate accumulation. Resource Conservation and Recycling. 1996;18:125–34. https://doi.org/10.1016/S0921-3449(96)01173-1
39. McGrath JW, Wisdom GB, McMullan G, Lrakin MJ, Quinn, JP. The purification and properties of phosphonoacetate hydrolase, a novel carbon-phosphorus bond-cleaving enzyme from Pseudomonas fluorescens 23F. European Journal of Biochemistry. 1995;234:225–30. https://doi.org/10.1111/j.1432-1033.1995.225_c.x
40. Bujacz B, Wieczorek P, Krzysko-Lupcka T, Golab Z, Lejczak B, Kavfarski P. Organophosphonate utilization by the wild-type strain of Penicillium notatum. Applied Environmental Microbiology. 1995;61:2905–10. https://doi.org/10.1128/AEM.61.8.2905-2910.1995
41. Krasilnikov M. On the role of soil bacteria in plant nutrition. Journal of General and Applied Microbiology. 1961;7:128–44. https://doi.org/10.2323/jgam.7.128
42. Hall JA, Pierson D, Ghosh S, Glick BR. Root elongation in various agronomic crops by the plant growth promoting rhizobacterium Pseudomonas putida GR12-2. Israel Journal of Plant Sciences. 1996;44:37–42. https://doi.org/10.1080/07929978.1996.10676631
43. Glick BR, Changping L, Sibdas G, Dumbroff EB. Early development of canola seedlings in the presence of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2. Soil Biological Biochemistry. 1997;29:1233–39. https://doi.org/10.1016/S0038-0717(97)00026-6
44. Kloepper JW, Lifshitz K, Schroth MN. Pseudomonas inoculants to benefit plant production. ISI Atlas of Science: Animal and Plant Sciences. 1988;60–64.
45. Kapulnik J, Gafny R, Okon Y. Effect of Azopirillum spp. inoculation on root development and NO-3 uptake in wheat (Titicum aestivum cv. Miriam) in hydroponic systems. Canadian Journal of Botany. 1985;63:627–31. https://doi.org/10.1139/b85-078
46. Broadbent P, Baker KF, Franks N, Holland J. Effect of Bacillus spp. on increased growth of seedlings in steamed and in nontreated soil. Phytopathology. 1977;67:1027–34. https://doi.org/10.1094/Phyto-67-1027
47. Burr TJ, Schroth MN, Suslow T. Increased potato yields by treatment of seedpieces with specific strains of Pseudomonas fluorescens and Pseudomonas putida. Phytopathology. 1978;68:1377–83. https://doi.org/10.1094/Phyto-68-1377
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31-12-2019
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Singh P, Banik RM. Effect of purified alkaline phosphatase from Bacillus licheniformis on growth of Zea mays L. Plant Sci. Today [Internet]. 2019 Dec. 31 [cited 2024 Dec. 22];6(sp1):583-9. Available from: https://horizonepublishing.com/journals/index.php/PST/article/view/676
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