Applied Microbiology and Biotechnology

, Volume 65, Issue 5, pp 559–565 | Cite as

Differential regulation and xenobiotic induction of tandem P450 monooxygenase genes pc-1 (CYP63A1) and pc-2 (CYP63A2) in the white-rot fungus Phanerochaete chrysosporium

Biotechnologically Relevant Enzymes and Proteins

Abstract

The two tandem P450 monooxygenase genes (pc-1 and pc-2) reported by us earlier in Phanerochaete chrysosporium were investigated for their regulation under nutrient-limited and nutrient-rich culture conditions. Transcript analysis based on real-time quantitative RT-PCR showed higher expression of pc-1 in defined low-nitrogen and pc-2 in defined high-nitrogen media, with maximum expression on day 4, indicating that the two genes, though tandemly linked, are regulated in a non-coordinate manner. Transcript levels of pc-1 and pc-2 were differentially influenced by the type of carbon source, incubation temperature, and oxygenation. Both genes were inducible by organic xenobiotic chemicals. Of the 42 xenobiotics tested in nutrient-rich cultures, pc-1 transcription was induced 2.12(±0.40)-fold to 6.27(±0.48)-fold in the presence of 19 compounds and pc-2 transcription was induced 2.00(±0.73)-fold to 29.39(±9.40)-fold in the presence of 22 compounds. Particularly, it is significant that both pc-1 and pc-2 are induced by polycyclic aromatic hydrocarbons (PAHs) of varying ring size, including naphthalene (4.35±0.09, 6.02±1.39), phenanthrene (2.82±0.12, 2.14±0.61), pyrene (3.93±0.01, 1.0±0.12), benzanthracene (1.67±0.03, 6.08±1.50), and benzo(a)pyrene (1.55±0.01, 5.54±2.75) respectively. This study constitutes the first report on the identification of P450 genes in a fungus that are responsive to environmentally significant pollutant chemicals (PAHs, DDT, long-chain alkyl phenols) and lignin derivatives.

References

  1. Anzenbacher P, Anzenbacherova E (2001) Cytochrome P450 and metabolism of xenobiotics. Cell Mol Life Sci 58:737–747PubMedGoogle Scholar
  2. Bumpus JA, Aust SD (1987) Biodegradation of environmental pollutants by the white rot fungus Phanerochaete chrysosporium: involvement of the lignin degrading system. BioEssays 6:166–170Google Scholar
  3. Cullen D (1997) Recent advances on the molecular genetics of ligninolytic fungi. J Biotechnol 53:273–289CrossRefPubMedGoogle Scholar
  4. Dosoretz CG, Dass SB, Reddy CA, Grethlein HE (1990) Protease-mediated degradation of lignin peroxidase in liquid cultures of Phanerochaete chrysosporium. Appl Environ Microbiol 56:3429–3434PubMedGoogle Scholar
  5. Fukuda H, Fujii T, Sukita E, Tazaki M, Nagahama S, Ogawa T (1994) Reconstitution of the isobutene forming reaction catalyzed by cytochrome P450 and P450 reductase from Rhodotorula minuta: decarboxylation with the formation of isobutene. Biochem Biophys Res Commun 201:516–522CrossRefPubMedGoogle Scholar
  6. Gold MH, Alic M (1993) Molecular biology of the lignin-degrading basidiomycete Phanerochaete chrysosporium. Microbiol Rev 57:605–622PubMedGoogle Scholar
  7. Keyser P, Kirk TK, Zeikus JG (1978) Ligninolytic enzyme system of Phanerochaete chrysosporium: synthesized in the absence of lignin in response to nitrogen starvation. J Bacteriol 135:790–797PubMedGoogle Scholar
  8. Kirk TK, Farrell RL (1987) Enzymatic “combustion”: the microbial degradation of lignin. Annu Rev Microbiol 41:465–505CrossRefPubMedGoogle Scholar
  9. Kullman SW, Matsumura F (1996) Metabolic pathways utilized by Phanerochaete chrysosporium for degradation of the cyclodiene pesticide endosulfan. Appl Environ Microbiol 62:593–600PubMedGoogle Scholar
  10. Kullman SW, Matsumura F (1997) Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium. Appl Environ Microbiol 63:2741–2746PubMedGoogle Scholar
  11. Lida T, Sumita T, Ohta A, Takagi M (2000) The cytochrome P450ALK multigene family of n-alkane-assimilating yeast, Yarrowia lipolytica: cloning and characterization of genes coding for new CYP52 family members. Yeast 16:1077–1087CrossRefPubMedGoogle Scholar
  12. Maloney AP, VanEtten HD (1994) A gene from the fungal plant pathogen Nectria haematococca that encodes the phytoalexin-detoxifying enzyme pisatin demethylase defines a new cytochrome P450 family. Mol Genet Genomics 243:506–514Google Scholar
  13. Nakashima A, Yoshida M, Nakayama K, Kato-Furuno A, Ueno M, Ushimaru T, Uritani M (2002) Genes for a nuclease and a protease are involved in the drastic decrease in cellular RNA amount in fission yeast cells during nitrogen starvation. J Biochem (Tokyo) 131:391–398Google Scholar
  14. Ohkuma M, Muraoka SI, Tanimoto T, Fujii M, Ohta A, Takagi M (1995) CYP52 (cytochrome P450alk) multigene family in Candida maltosa: identification and characterization of eight members. DNA Cell Biol 14:163–173PubMedGoogle Scholar
  15. Orth AB, Tien M (1995) Biotechnology of lignim degradation. In: Kuck (ed) The mycota II genetics and biotechnology. Springer, Berlin Heidelberg New York, pp 287–302Google Scholar
  16. Paszczynski A, Crawford RL (1995) Potential for bioremediation of xenobiotic compounds by the white rot fungus Phanerochaete chrysosporium. Biotechnol Prog 11:368–379Google Scholar
  17. Reddy CA (1995) The potential of white rot fungi for the treatment of pollutants. Curr Opin Biotechnol 6:320–328CrossRefGoogle Scholar
  18. Sutherland JB, Selby AL, Freeman JP, Evans FE, Cerniglia CE (1991) Metabolism of phenanthrene by Phanerochaete chrysosporium. Appl Environ Microbiol 57:3310–3316PubMedGoogle Scholar
  19. Van den Brink HJM, Van Gorcom RFM, Van den Hondel CAMJJ, Punt PJ (1998) Cytochrome P450 enzyme systems in fungi. Fungal Genet Biol 23:1–17CrossRefPubMedGoogle Scholar
  20. Van Gorcom RFM, Boschloo JG, Kuijvenhoven A, Lange J, Van Vark AJ, Bos CJ, Van Balken JAM, Pauwels PH, Van den Hondel CAMJJ (1990) Isolation and molecular characterization of the benzoate-para-hydroxylase gene (bphA) of Aspergillus niger: a member of a new family of the cytochrome P450 superfamily. Mol Genet Genomics 223:192–197Google Scholar
  21. Yadav JS, Doddapaneni H (2003) Genome-wide expression profiling and xenobiotic inducibility of P450 monooxygenase genes in the white rot fungus Phanerochaete chrysosporium. In: Anzenbacher P, Hudecek J (eds) Cytochrome P450 biochemistry, biophysics and drug metabolism. (13th international conference on cytochrome P450) Monduzzi, Bologna, pp 333–340Google Scholar
  22. Yadav JS, Loper JC (1999) Multiple P450alk (cytochrome P450 alkane hydroxylase) genes from the halotolerant yeast Debaryomyces hansenii. Gene 226:139–146CrossRefPubMedGoogle Scholar
  23. Yadav JS, Loper JC (2000) Cytochrome P450 oxidoreductase gene and its differentially terminated cDNAs from the white rot fungus Phanerochaete chrysosporium. Curr Genet 37:65–73CrossRefPubMedGoogle Scholar
  24. Yadav JS, Lawrence D, Nuck B, Federle T, Reddy CA (2001) Biotransformation of linear alkylbenzene sulfonate (LAS) by Phanerochaete chrysosporium: oxidation of alkyl side-chain. Biodegradation 12:443–453CrossRefPubMedGoogle Scholar
  25. Yadav JS, Soellner MB, Loper JC, Mishra PK (2003) Tandem cytochrome P450 monooxygenase genes and splice variants in the white rot fungus Phanerochaete chrysosporium: cloning, sequence analysis, and regulation of differential expression. Fungal Genet Biol 38:10–21CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Molecular Toxicology Division, Department of Environmental HealthUniversity of Cincinnati, College of MedicineCincinnatiUSA

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