Bio/Technology
9, 987 - 990 (1991)
doi:10.1038/nbt1091-987
Heterologous Production of a Ligninolytic Enzyme: Expression of the Phlebia Radiata Laccase Gene in Trichoderma ReeseiMarkku Saloheimo1, *
& Marja-Leena Niku-Paavola1
1VTT, Biotechnical Laboratory, P.O. Box 202, SF-02151 Espoo, Finland.
*Corresponding author. We have expressed a gene encoding the ligninolytic laccase enzyme of the white−rot fungus Phlebia radiata in the soft rot fungus Trichoderma reesei under the promoter of the major cellulase gene (cbh1). Constructions from the laccase cDNA or the chromosomal gene were equivalent in expression, yielding 20 mg/l of secreted active laccase in small−scale fermentations. The recombinant enzyme has a similar molecular weight, antigenic properties and specific activity as the Phlebia laccase and is analogous to the Phlebia enzyme in removing monomeric lignin−related compounds from residual lignin of the pulping process, thus demonstrating its potential for industrial applications.
REFERENCES
- Kirk, T.K. and Farrell, R.L. 1987. Enzymatic "combustion": The microbial degradation of lignin. Ann. Rev. Microbiol. 41: 465−505. | Article | ISI | ChemPort |
- Niku-Paavola, M-L., Karhunen, E., Salola, P. and Raunio, V. 1988. Ligninolytic enzymes of the white-rot fungus Phlebia radiata. Biochem. J. 254: 877−884. | PubMed | ChemPort |
- Karhunen, E., Kantelinen, A. and Niku-Paavola, M.-L. 1990. Mn-dependent peroxidase from the lignin-degrading white-rot fungus Phlebia radiata. Arch. Biochem. Biophys. 279: 25−31. | PubMed | ChemPort |
- Karhunen, E., Niku-Paavola, M.-L., Viikari, L., Haltia, T., van de Meer, R.A. and Duine, J.A.1990. A novel combination of prosthetic groups in a fungal laccase: PQQ and two copper atoms. FEBS Letts. 267: 6−8. | Article | ChemPort |
- Saloheimo, M., Barajas, V., Niku-Paavola, M.-L. and Knowles, J.K.C. 1989. A lignin peroxidase-encoding cDNA from the white-rot fungus Phlebia radiata: characterization and expression in Trichoderma reesei. Gene 85: 343−351. | Article | PubMed | ChemPort |
- Saloheimo, M., Niku-Paavola, M.-L. and Knowles, J.K.C. 1991. Isolation and structural analysis of the laccase gene from the lignin-degrading fungus Phlebia radiata. J. Gen. Microbiol. In press.
- Saunders, G., Picknett, T.M., Tuite, M.F. and Ward, M. 1989. Heterologous gene expression in filamentous fungi. Trends in Biotechnol. 7: 283−287. | ISI | ChemPort |
- Harkki, A., Uusitalo, J., Bailey, M., Penttilä, M. and Knowles, J.K.C. 1989. A novel fungal expression system: secretion of active calf chymosin from the filamentous fungus Trichoderma reesei. Bio/Technology 7: 596−603. | ISI | ChemPort |
- Uusitalo, J.M., Nevalainen, K.M.H., Harkki, A.M., Knowles, J.K.C. and Penttilä, M.E. 1991. Enzyme production by recombinant Trichoderma reesei strains. J. Biotechnol. 17: 35−50. | Article | PubMed | ChemPort |
- Mattern, I.E., Punt, P.J., Unkles, S., Pouvels, P.H. and van den Hondel, C.A.M.J.J. 1987. Transfomation of Aspergillus oryzae. In: Abstracts of the 19th Lunteren Lectures on Molecular Genetics of Yeasts and Filamentous Fungi and its Impact on Biotechnology, p. 34. Lunteren, The Netherlands.
- Hynes, M.J., Corrick, C.M. and King, J.A. 1983. Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol. Cell. Biol. 3: 1430−1439. | PubMed | ISI | ChemPort |
- Ballance, D.J. 1986. Sequences important for gene expression in filamentous fungi. Yeast 2: 229−236. | PubMed | ISI | ChemPort |
- Penttilä, M., Lehtovaara, P., Nevalainen, H., Bhikhabhai, R. and Knowles, J.K.C. 1986. Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene. Gene 45: 253−263. | Article | PubMed | ChemPort |
- Kuan, I. and Tien, M. 1989. Phosphorylation of lignin peroxidases from Phanerochaete chrysosporium. J. Biol. Chem. 264: 20350−20355. | PubMed | ChemPort |
- Davis, S. and Burns, R.G. 1990. Decolorization of phenolic effluents by soluble and immobilized phenol oxidases. Appl. Microbiol. Biotechnol. 32: 721−726. | ChemPort |
- Farrell, R., Gelep, P., Anilionis, A., Javaherian, K., Maione, T.E., Rushe, J.R., Sadownick, B.A. and Jackson, J.A. 1987. Recombinant ligninase. European Patent Application No. 87810516.2.
- Kojima, Y., Tsukuda, Y., Kawai, Y., Tsukamoto, A., Sugiura, J. and Sakaino, M., 1990. Cloning, sequence analysis, and expression of ligninolytic phenoloxidase genes of the white-rot basidiomycete Coriolus hirsutus. J. Biol. Chem. 265: 15224−15230. | PubMed | ChemPort |
- Niku-Paavola, M.-L., Karhunen, E., Kantelinen, A., Viikari, L., Lundell, T. and Hatakka, A. 1990. The effect of culture conditions on the production of lignin modifying enzymes by the white-rot fungus Phlebia radiata. J. Biotechnol. 13: 211−221. | Article | ChemPort |
- Ward, M., Wilson, L.J., Kodama, K.H., Rey, M.W. and Berka, R.M. 1990. Improved production of chymosin in Aspergillus by expression as a glucoamylase-chymosin fusion. Bio/Technology 8: 435−440. | PubMed | ISI | ChemPort |
- Lamsa, M. and Bloebaum, P. 1990. Mutation and screening to increase chymosin yield in a genetically-engineered strain of Aspergillus awamori. J. Industr. Microbiol. 5: 229−238. | ChemPort |
- Sheir-Neiss, G. and Montenecourt, B.S. 1984. Characterization of the secreted cellulases of Trichoderma reesei wild type and mutants during controlled fermentations. Appl. Microbiol. Biotechnol. 20: 46−53. | ChemPort |
- Yannish-Perron, C., Vieira, J. and Messing, J. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mpl8 and pUC19 vectors. Gene 33: 103−119. | Article | PubMed |
- Penttilä, M., Nevalainen, H., Rättö, M., Salminen, E. and Knowles, J. 1987. A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei.Gene 61: 155−164. | Article | PubMed | ISI | ChemPort |
- Raeder, U. and Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1: 17−20. | ISI | ChemPort |
- Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J. and Rutter, W.J. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochem. J. 18: 5294−5299. | ChemPort |
- Bailey, M.J. and Ojamo, H. 1990. Selective concentration of polygalacturonase and beta-D-glucosidase of Aspergillus niger culture filtrate using mineral absorbents. Bioseparation 1: 133−139. | ChemPort |
- Bradford, M.M. 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. | Article | PubMed | ISI | ChemPort |
- Niku-Paavola, M.-L., Raaska, L. and Itävaara, M. 1990. Detection of white-rot fungi by a nontoxic stain. Mycol. Res. 94: 27−31.
- Alen, R. and Vikkula, A. 1989. Formation of lignin monomers during alkaline delignification of softwood. Holzforschung 43: 397−400. | ChemPort |
|
