Screening for and characterization of phospholipase A1 hypersecretory mutants of Tetrahymena thermophila
We have described a procedure for the isolation of mutants of Tetrahymena thermophila with hyperscretion of phospholipase A1 (PLA1). Using random chemical mutagenesis, uniparental cytogamy, genetic crossing and a new, fast and effective screening procedure, four PLA1-hypersecretory mutants were isol...
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Springer Verlag
2000
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| Acceso en línea: | Registro en Scopus DOI Handle Registro en la Biblioteca Digital |
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| LEADER | 09424caa a22009497a 4500 | ||
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| 001 | PAPER-2256 | ||
| 003 | AR-BaUEN | ||
| 005 | 20230518203139.0 | ||
| 008 | 190411s2000 xx ||||fo|||| 00| 0 eng|d | ||
| 024 | 7 | |2 scopus |a 2-s2.0-0033802432 | |
| 024 | 7 | |2 cas |a phosphatidylcholine, 55128-59-1, 8002-43-5; phospholipase A1, 9043-29-2 | |
| 040 | |a Scopus |b spa |c AR-BaUEN |d AR-BaUEN | ||
| 030 | |a AMBID | ||
| 100 | 1 | |a Hartmann, M. | |
| 245 | 1 | 0 | |a Screening for and characterization of phospholipase A1 hypersecretory mutants of Tetrahymena thermophila |
| 260 | |b Springer Verlag |c 2000 | ||
| 270 | 1 | 0 | |m Hartmann, M.; Inst. fur Allgemeine Zool./Genetik, Westfalische Wilhelms-Universitat, 48149 Munster, Germany; email: tiedtke@uni-muenster.de |
| 506 | |2 openaire |e Política editorial | ||
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| 504 | |a Bruns, P.J., Brussard, T.B., Kavka, A.B., Isolation of homozygous mutant after induced selffertilization in Tetrahymena (1976) Proc Natl Acad Sci USA, 73, pp. 3243-3247 | ||
| 504 | |a Chelius, M.K., Wodzinski, R.J., Strain improvement of Aspergillus niger for phytase production (1994) Appl Microbiol Biotechnol, 41, pp. 79-83 | ||
| 504 | |a Cole, E.S., Bruns, P.J., Uniparental cytogamy: A novel method for bringing micronuclear mutations of Tetrahymena into homozygous macronuclear expression with precocious sexual maturity (1992) Genetics, 132, pp. 117-131 | ||
| 504 | |a Dimond, R.L., Knecht, D.A., Jordan, K.B., Burns, R.A., Livi, G.P., Secretory mutants in the cellular slime mold Dictyostelium discoideum (1983) Methods Enzymol, 96, pp. 815-828 | ||
| 504 | |a Ebert, D.L., Jordan, K.B., Dimond, R.L., Lysosomal enzyme secretory mutants of Dictyostelium discoideum (1990) J Cell Sci, 96, pp. 491-500 | ||
| 504 | |a Florin-Christensen, J., Florin-Christensen, M., Knudsen, J., Rasmussen, L., Cytolytic activity released from Tetrahymena (1985) J Protozool, 32, pp. 657-660 | ||
| 504 | |a Florin-Christensen, J., Florin-Christensen, M., Rasmussen, L., Knudsen, J., Hansen, H.O., Phospholipase A1 and triacylglycerol lipase: Two novel enzymes from Tetrahymena culture medium (1986) Comp Biochem Physiol, 85 B, pp. 149-155 | ||
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| 504 | |a Hunseler, P., Scheidgen-Kleyboldt, G., Tiedtke, A., Isolation and characterization of a mutant of Tetrahymena thermophila blocked in secretion of lysosomal enzymes (1987) J Cell Sci, 88, pp. 47-55 | ||
| 504 | |a Jain, S., Parriche, M., Durand, H., Tiraby, G., Production of polysaccharidases by a cellulase-pectinase hyperproducing mutants (Po16) of Penicillium occitanis (1990) Enzyme Microb Technol, 12, pp. 691-696 | ||
| 504 | |a Kim, K.M., Rhee, J.S., Simple and rapid screening methods for microorganisms with phospholipase A1, A2 and C activities (1994) Biotechnol Tech, 8, pp. 635-638 | ||
| 504 | |a Kiy, T., Tiedtke, A., Lysosomal enzymes produced by immobilized Tetrahymena thermophila (1991) Appl Microbiol Biotechnol, 35, pp. 14-18 | ||
| 504 | |a Kiy, T., Tiedtke, A., Continuous high-cell-density fermentation of the ciliated protozoon Tetrahymena in a perfused bioreactor (1992) Appl Microbiol Biotechnol, 38, pp. 141-146 | ||
| 504 | |a Kiy, T., Tiedtke, A., Mass cultivation of Tetrahymena thermophila yielding high cell densities and short generation times (1992) Appl Microbiol Biotechnol, 37, pp. 576-579 | ||
| 504 | |a Kiy, T., Tiedtke, A., Protozoen in der biotechnologie: Ciliaten auf neuen wegen (1993) BioEngineering, 9, pp. 22-36 | ||
| 504 | |a Kiy, T., Hoersch, B., Marquardt, R., (1995), Process for the preparation for glycosides using glycosidases from ciliates. Eur Pat Appl EP 725,144 (C1 C12P19/14), 7 Aug 1996. DE Appl 19.503.649.6, 6 Feb 1995; Kiy, T., Scheidgen-Kleyboldt, G., Tiedtke, A., Production of lysosomal enzymes by continuous high-cell-density fermentation of the ciliated protozoon Tetrahymena thermophila in a perfused bioreactor (1996) Enzyme Microb Tech, 118, pp. 268-274 | ||
| 504 | |a Muller, M., Secretion of acid hydrolases and its intracellular source in Tetrahymena pyriformis (1972) Cell Biol, 52, pp. 478-487 | ||
| 504 | |a Munro, I.G., Protozoa as sources of commercially produced enzymes - A review (1985) Process Biochem, 20, pp. 139-144 | ||
| 504 | |a Rothstein, T.L., Blum, J.J., Lysosomal physiology in Tetrahymena II. Effect of culture age and temperature on the extracellular release of three acid hydrolases (1974) J Protozool, 21, pp. 163-168 | ||
| 504 | |a Tiedtke, A., Purification and properties of secreted N-acetyl-β-D-hexosaminidase of Tetrahymena thermophila (1983) Comp Biochem Physiol, 75 B, pp. 239-243 | ||
| 504 | |a Vandamme, E.J., The search for novel microbial fine chemicals, agrochemicals and biopharmaceuticals (1994) J Biotechnol, 37, pp. 89-108 | ||
| 520 | 3 | |a We have described a procedure for the isolation of mutants of Tetrahymena thermophila with hyperscretion of phospholipase A1 (PLA1). Using random chemical mutagenesis, uniparental cytogamy, genetic crossing and a new, fast and effective screening procedure, four PLA1-hypersecretory mutants were isolated. The screening procedure is based on the formation of a halo appearing around cylindrical holes in a lecithin-containing agar plate filled with cell-free supernatants. About 3,940 clones were tested with this procedure in primary screening for hypersecretory features, of which 60 putative hypersecretory mutants were isolated, subcloned and tested in a secondary screening. Of these, four selected mutants showed 1.8-2.2 more PLA1 activity in the cell-free supernatants compared to the wild-type strain CU 438.1. Hypersecretion was only observable for PLA1; no increased activity for two other lysosomal enzymes could be detected. These hypersecretory mutants of T. thermophila can be very useful for increasing the yield of PLA1 in fermentation processes. This is particularly relevant because, in contrast to other phospholipases, PLA1 is not available on the commercial market for fine chemicals and little is known about the role of PLA1 in cell signaling and metabolism. |l eng | |
| 536 | |a Detalles de la financiación: European Commission | ||
| 536 | |a Detalles de la financiación: Acknowledgements We thank Prof. Leif Rasmussen for critical reading of and helpful comments on the manuscript. The technical instructions on the radiochemical enzyme assay by Dr. Jorge Florin-Christensen is acknowledged. This study was supported by the EC grant No. CI1-CT94-0026 to A.T. and M.F.-C. | ||
| 593 | |a Institut für Allgemeine Zoologie und Genetik, Westfälische Wilhelms-Universität, 48149 Münster, Germany | ||
| 593 | |a Laboratory of Molecular Biology, Department of Biological Chemistry, Ciudad Universitaria, 1428 Buenos Aires, Argentina | ||
| 593 | |a Institute of Neuroscience (INEUCI), Ciudad Universitaria, Pabellon 2, 1428 Buenos Aires, Argentina | ||
| 690 | 1 | 0 | |a LYSOSOME ENZYME |
| 690 | 1 | 0 | |a PHOSPHATIDYLCHOLINE |
| 690 | 1 | 0 | |a PHOSPHOLIPASE A1 |
| 690 | 1 | 0 | |a ARTICLE |
| 690 | 1 | 0 | |a BACTERIUM ISOLATION |
| 690 | 1 | 0 | |a BACTERIUM MUTANT |
| 690 | 1 | 0 | |a CHEMICAL MUTAGENESIS |
| 690 | 1 | 0 | |a ENZYME ACTIVITY |
| 690 | 1 | 0 | |a ENZYME ANALYSIS |
| 690 | 1 | 0 | |a ENZYME RELEASE |
| 690 | 1 | 0 | |a FERMENTATION |
| 690 | 1 | 0 | |a MOLECULAR CLONING |
| 690 | 1 | 0 | |a NONHUMAN |
| 690 | 1 | 0 | |a SCREENING |
| 690 | 1 | 0 | |a STRAIN DIFFERENCE |
| 690 | 1 | 0 | |a SUPERNATANT |
| 690 | 1 | 0 | |a TETRAHYMENA THERMOPHILA |
| 690 | 1 | 0 | |a BACTERIA (MICROORGANISMS) |
| 690 | 1 | 0 | |a PROKARYOTA |
| 690 | 1 | 0 | |a PROTOZOA |
| 690 | 1 | 0 | |a TETRAHYMENA THERMOPHILA |
| 690 | 1 | 0 | |a TETRAHYMENA THERMOPHILA |
| 700 | 1 | |a Guberman, A. | |
| 700 | 1 | |a Florin-Christensen, M. | |
| 700 | 1 | |a Tiedtke, A. | |
| 773 | 0 | |d Springer Verlag, 2000 |g v. 54 |h pp. 390-396 |k n. 3 |p Appl. Microbiol. Biotechnol. |x 01757598 |w (AR-BaUEN)CENRE-173 |t Applied Microbiology and Biotechnology | |
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