Research Paper | Published:

Construction of a 7-Aminocephalosporanic Acid (7ACA) Biosynthetic Operon and Direct Production of 7ACA in Acremonium Chrysogenum

Bio/Technologyvolume 9pages188191 (1991) | Download Citation

Subjects

Abstract

We have used cDNA encoding D-amino acid oxidase, and genomic DNA encoding cephalosporin acylase from Fusarium solani and Pseudomonas diminuta, respectively, to construct a novel hybrid 7-aminocephalosporanic acid (7ACA) biosynthetic operon under the control of regulatory elements from the alkaline protease gene of Acremonium chrysogenum. Transformants of A. chrysogenum BC2116, a high cephalosporin-producing strain, containing this operon, synthesized and secreted low levels of 7ACA. Although the amounts are not yet commercially significant, this represents the first microbial production of 7ACA and demonstrates the feasibility of introducing new biosynthetic capabilities into industrial microorganisms by combining fungal and bacterial genes.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Bunnel, C.A., Luke, W.D. and Perry, F.M. 1986. Industrial manufacture of cephalosporins, p.255–284. In: Beta-lactam Antibiotics for Clinical Use. S.F. Queener, J.A. Webber, and S.W. Queener (Eds.). Marcel Dekker, New York.

  2. 2

    Tsuzuki, K., Komatsu, K.-I., Ichikawa, S. and Shibuya, Y. 1989. Enzymatic synthesis of 7-aminocephalosporanic acid (7-ACA). Nippon Nogeikagaku Kaishi 63: 1847–1853.

  3. 3

    Malpartida, F. and Hopwood, D.A. 1984. Molecular cloning of the whole biosynthetic pathway of a Streptomyces antibiotic and its expression in a heterologous host. Nature 309: 462–464.

  4. 4

    Chen, C.W., Lin, H.-F., Kuo, C.L., Tsai, H.-L. and Tsai, J.F.-Y. 1988. Cloning and expression of a DNA sequence conferring cephamycin C production. Bio/Technology 6: 1222–1224.

  5. 5

    Smith, D.J., Burnham, M.K.R., Edward, J., Earl, A.J. and Turner, G. 1990. Cloning and heterologous expression of the penicillin biosynthetic gene cluster from Penicillium chrysogenum. Bio/Technology 8: 39–41.

  6. 6

    Isogai, T., Ono, H. and Kojo, H. 1990. D-Amino acid oxidase. Eur. Pat. Appl. EP 364,275 (18 Apr 1990).

  7. 7

    Isogai, T., Ono, H., Ishitani, Y., Kojo, H., Ueda, Y. and Kohsaka, M. 1990. Structure and expression of cDNA for D-amino acid oxidase active against cephalosporin C from Fusarium solani. J. Biochem. 108: 1063–1069.

  8. 8

    Szwajcer, E. and Mosbach, K. 1985. Isolation and partial characterization of a D-amino acid oxidase active against cephalosporin C from the yeast Trigonopsis variabilis. Biotechnology Letters 7: 1–7.

  9. 9

    Matsuda, A., Matsuyama, K., Yamamoto, K., Ichikawa, S. and Komatsu, K.-I. 1987. Cloning and characterization of the genes for two distinct cephalosporin acylases from a Pseudomonas strain. J. Bacteriol. 169: 5815–5820.

  10. 10

    Matsuda, A., Toma, K. and Komatsu, K.-I. 1987. Nucleotide sequences of the genes for two distinct cephalosporin acylases from a Pseudomonas strain. J. Bacteriol. 169: 5821–5826.

  11. 11

    Isogai, T., Fukagawa, M., Kojo, H., Kohsaka, M., Aoki, H. and Imanaka, H. 1991. Cloning and nucleotide sequence of the complementary and genomic DNAs for the alkaline protease from Acremonium chrysogenum. Agric. Biol. Chem. In press.

  12. 12

    Yagi, J., Yano, T., Kubochi, Y., Hattori, S., Ohashi, M., Sakai, H., Jomon, K. and Ajisaka, M. 1972. Studies on alkaline proteinase from Cephalosporium. J. Ferment. Technol. 50: 592–599.

  13. 13

    Skatrud, P.L., Queener, S.W., Carr, L.G. and Fisher, D.L. 1987. Efficient integrative transformation of Cephalosporium acremonium. Curr. Genet. 12: 337–348.

  14. 14

    Skatrud, P.L., Tietz, A.J., Ingolia, T.D., Cantwell, C.A., Fisher, D.L., Chapman, J.L. and Queener, S.W. 1989. Use of recombinant DNA to improve production of cephalosporin C by Cephalosporium acremonium. Bio/Technology 7: 477–485.

  15. 15

    Gritz, L. and Davies, J. 1983. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25: 179–188.

  16. 16

    van den Elzen, P.J.M., Townsend, J., Lee, K.Y. and Bedbrook, J.R. 1985. A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Molecular Biology 5: 299–302.

  17. 17

    Takeshita, S., Sato, M., Toba, M., Masahashi, W. and HashimotoGotoh, T. 1987. High-copy-number and low-copy-number plasmid vectors for lacZα-complementation and chloramphenicol- or kanamycin-resistance selection. Gene 61: 63–74.

  18. 18

    Shen, Y.-Q., Wolfe, S. and Demain, A.L. 1986. Levels of isopenicillin N synthetase and deacetoxycephalosporin C synthetase in Cephalosporium acremonium producing high and low levels of cephalosporin C. Bio/Technology 4: 61–64.

  19. 19

    Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, New York.

  20. 20

    Isogai, T., Yoshida, M., Tanaka, M. and Aoki, H. 1987. Transformation of Acremonium chrysogenum and Saccharomyces cerevisiae using an antibiotic resistance marker. Agric. Biol. Chem. 51: 2321–2329.

  21. 21

    Samson, S.M., Belagaje, R., Blankenship, D.T., Chapman, J.L., Perry, D., Skatrud, P.L., VanFrank, R.M., Abraham, E.P., Baldwin, J.E., Queener, S.W. and Ingolia, T.D. 1985. Isolation, sequence determination and expression in Escherichia coli of the isopenicillin N synthetase gene from Cephalosporium acremonium. Nature 318: 191–194.

Download references

Author information

Author notes

  1. Takao Isogai: Corresponding author.

Affiliations

  1. Exploratory Research Laboratories, Fujisawa Pharmaceutical Co. Ltd, Tokodai, Tsukuba, Ibaraki, 300-26, Japan

    • Takao Isogai
    • , Masao Fukagawa
    • , Ichiro Aramori
    • , Morita Iwami
    • , Hitoshi Kojo
    •  & Takaharu Ono
  2. Research and Development Group, Fujisawa Pharmaceutical Co. Ltd, Kashima, Yodogawa-ku, Osaka, 532, Japan

    • Yoshio Ueda
    • , Masanobu Kohsaka
    •  & Hiroshi Imanaka

Authors

  1. Search for Takao Isogai in:

  2. Search for Masao Fukagawa in:

  3. Search for Ichiro Aramori in:

  4. Search for Morita Iwami in:

  5. Search for Hitoshi Kojo in:

  6. Search for Takaharu Ono in:

  7. Search for Yoshio Ueda in:

  8. Search for Masanobu Kohsaka in:

  9. Search for Hiroshi Imanaka in:

About this article

Publication history

Received

Accepted

Issue Date

DOI

https://doi.org/10.1038/nbt0291-188

Further reading