Browse > Article
http://dx.doi.org/10.5657/fas.2003.6.1.020

Cloning and Expression of Escherichia coli K13 Phytase Gene (appA13) Isolated from Seawater  

Kim Young-Ok (Biotechnology Research Center, National Fisheries Research and Development Institute)
Kim Han-Woo (Biotechnology Research Center, National Fisheries Research and Development Institute)
Lee Jung-Ho (Biotechnology Research Center, National Fisheries Research and Development Institute)
Kim Kyung-Kil (Biotechnology Research Center, National Fisheries Research and Development Institute)
Lee Jong-Yun (Biotechnology Research Center, National Fisheries Research and Development Institute)
Publication Information
Fisheries and Aquatic Sciences / v.6, no.1, 2003 , pp. 20-26 More about this Journal
Abstract
A bacterial strain was isolated from seawater to screen for phytase activities. A colony had the highest activity and was identified as an Escherichia coli strain. Using primers derived from E. coli acid phosphatase appA sequence, we cloned a 1,495 bp DNA fragment connected with the pGEM-T vector. It was over-expressed under lac promoter combined with its native promoter in E. coli $DH5\alpha$. The expression of the phytase gene occurred during late exponential growth and the intracellular phytase production was 16.9 units/ml. The yield of recombinant phytate was 412-fold higher than that of wild type E. coli K13.
Keywords
Escherichia coli; Phytate; Phytase; Expression;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Rimbach, G., H.J. Ingelmann and J. Pallauf. 1994. The role of phytase in the dietary bioavailability of minerals and trace elements. Ernahrungsforschung, 39, 1- 10
2 Rodriguez, E.Y., Y. Han and X.G. Lei. 1999. Cloning, sequencing and expression of Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon. Biochern. Biophys. Res. Comm., 257, 117-123   DOI   ScienceOn
3 Shimizu, M. 1992. Purification and characterization of phytase from Bacillus subtilis (natto) N-77. Biosci. Biotechnol. Biochem, 56, 1266-1269   DOI
4 Ullah, A.A.H. and D.M. Gibson. 1986. Purification and characterization of acid phosphatase from cotyledons for germinating soybean seeds. Arch. Biochem. Biophys., 260, 514- 520
5 Van Hartingseldt, W., C.M.J. van Zeijl, G.M. Harteveld, R.J. Gouka, M.E.G. Suykerbuyk, R.G.M. Luiten, P.A van Paridon, G.C.M. Selten, A.E. Veenstra, R.F.M. van Gorcom and C.A.M.J.J. van den Hondel. 1993. Cloning, characterization and overexpression of the phytaseencoding gene (phyA) of Aspergillus niger. Gene, 127, 87-94   DOI   ScienceOn
6 Wodzinski, R.J. and A.H.J. UIIah. 1996. Phytase. Adv. Appl. Microbiol., 42, 263- 302   DOI
7 Yamada, K, Y. Minoda and K. Yamada. 1968. Phytase from Aspergillus terrus. Part 1. Production, purification, and some general properties of the enzyme. Agric. Biol, Chem., 32, 1275-1283   DOI
8 Yoon, S.J., Y.J. Choi, HK Min, K.K Cho, j.W. Kim, S.C. Lee and Y.H. Jung. 1996. Isolation and identification of phytase-producing bacterium, Enterobacter sp. 4 and enzymatic properties of phytase enzyme. Enzyme Microb. Technol., 18, 449-454   DOI   ScienceOn
9 Greaves, M.P., G. Anderson and D.M. Webley. 1967. The hydrolysis of inositol phosphates by Aerobacter aerogenes. Biochim. Biophys. Acta., 132, 412-418   DOI   ScienceOn
10 Greiner, R., U. Konietzny and K.D. Jany. 1993. Purification and characterization of two phytases from Escherichia coli. Arch. Biochem. Biophys., 303, 107-113   DOI   ScienceOn
11 Holt, J.G., N.R. Krieg, P.H.A Sneath, J.T. Staley and S.T. Williams. 1994. Bergey's Manual of Determinative Bacteriology, 9th ed. Baltimore, MD: Williams and Wilkins, pp. 175-289
12 Mitchell, D.B., K. Voge, B.J. Weimann, L. Pasamontes and A.P.G.M. van Loon. 1997. The phytase subfamily of histidine acid phosphatase: isolation of genes for two novel phytases from the fungi Aspergillus terreus and Myceliophthora thennophila. Microbiology, 143, 245-252   DOI   ScienceOn
13 Irving, G.C. and D.J. Cosgrove. 1971. Inositol phosphate phosphatases of microbiological origin. Some properties of a partially purified bacterial (Pseudomonas sp.) phytase. Aust. J. Biol. Sci., 24, 547- 557
14 Kim, Y.O., H.K Kim, K.S. Bae, J.H. Yu and T.K. Oh. 1998a. Purification and properties of a thermostable phytase from Bacillus sp. DS11. Enzyme Microb. Technol., 22, 2 -7   DOI   ScienceOn
15 Kim, Y.O., J.K. Lee, H.K Kim, J.H. Yu and T.K Oh. 1998b. Cloning of thermostable phytase from Bacillus sp. DS11 and its overexpression in E. coli. FEMS Microbiol. Lett., 162, 185-191   DOI   ScienceOn
16 Pasamontes, L.M. Haiker, M. Henriquez-Huecas, D.B. Mitchell and A.P.G.M. van Loon. 1997. Cloning of the phytases from Emericella nidulans and the thermophilic fungus Talaromyces thennophilus. Biochim. Biophys. Acta., 1353, 217-223   DOI   ScienceOn
17 Reddy, N.R., S.K Sathe and D.K Salunkhe. 1982. Phytates in legumes and cereals. Adv. Food Res., 28, 1-92   DOI
18 Gibsin, D.M. and A.H.J. Ullah. 1988. Purification and characterization of phytase from cotyledons of germinating soybean seeds. Arch. Biochem. Biophys., 260, 503- 513   DOI   ScienceOn
19 Dassa, J., C. Marek and P.L. Boquet. 1990. The complete nucleotide sequence of the Escherichia coli gene appA reveals significant homology between pH 2.5 acid phosphatase and glucose-l-phosphatase, J. Bacteriol., 172, 5497- 5500   DOI
20 Fiske, C.H. and Y.P. Subbarow. 1925. The colormetric determination of phosphorous. J. Biol. Chem., 66, 375-410