Applications of Molecular Biology Techniques to the Remediation of Hazardous Waste

  • Burt D. Ensley
Part of the Industry-University Cooperative Chemistry Program Symposia book series (IUCC)


There are a growing number of applications of molecular biology tools to the manipulation of microorganisms involved in the degradation of hazardous compounds. These tools, including pathway cloning, polymerase chain reaction gene amplification, the use of hybrid promoter and regulatory sequences, broad host range and high copy number plasmids, DNA sequencing and synthesis methods, and enzyme recruitment can all be used to construct highly sophisticated biological catalysts that offer potentially superior performance in degradation. The use of genetically engineered microorganisms for the degradation of hazardous molecules is not yet a widely accepted approach. At the same time, there are many advantages in using an organism containing a precisely engineered pathway that encodes enzymes capable of rapidly attacking highly recalcitrant or toxic molecules. Some of the potential benefits in the application of existing molecular biology techniques to the construction of hazardous waste degrading organisms are described in this chapter.


Catabolite Repression Protein Engineering Molecular Biology Technique Increase Reaction Rate High Level Synthesis 
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  1. 1.
    W. N. Burnette, V. L. Marr and W. Cieplak, Direct expression of Bordetalla pertussis toxin subunits to high levels in Escherichia coli. Bio /Technology. 6:699–706 (1988).Google Scholar
  2. 2.
    P. Maxwell, J. Hsieh, and J. Fieschko, Stabilization of a mutant microorganism population, European patent, EP0098750, (1984).Google Scholar
  3. 3.
    R. V. Subba-Rao, H. E. Rueben, and M. Alexander, Kinetics and extent of mineralization of organic chemicals at trace levels in fresh water and sewage, Appl. and Environ. Microbiol. 43:1139–1150 (1982).Google Scholar
  4. 4.
    M. Alexander, Ecologie constraints on genetic engineering, in: “Genetic Control of Environmental Pollutants,” G.S. Ullman, and A. Hollaender, Plenum Press, New York (1984).Google Scholar
  5. 5.
    R. B. Winter, K. M. Yen and B. D. Ensley, Efficient degradation of trichloroethylene by a recombinant Escherichia coli. Bio/Technology. 7:282–285 (1989).CrossRefGoogle Scholar
  6. 6.
    C. M. Serdar, B. C. Murdock, and M. F. Rohde, Parathion hydrolase gene from Pseudomonas dimunitamg: subcloningcomplete nucleotide sequence and expression of a mature portion of the enzyme in Escherichia coli. Bio/Technology, 7:1151–1155 (1989).Google Scholar
  7. 7.
    K. Nagahari, Deletion plasmids from transformants of Pseudomonas aeuroginosa trp cells RSF1010 trp hybrid plasmid and high levels of enzyme activity from the gene on the plasmid, J. Bacteriol., 136:312–318 (1978).PubMedGoogle Scholar
  8. 8.
    D. B. Janssen, F. Pries, J. Ploeg, B. Kavemier, P. Terpstra, and B. Witholt, Cloning of 1, 2-dichloroethane degradation genes of Xanthobacter autotrophicus GJ10 and expression and sequencing of the dhlA gene, J. Bacteriol., 171:6791–6799 (1989).PubMedGoogle Scholar
  9. 9.
    B. D. Ensley, T. D. Osslund, M. Joyce, and M. J. Simon, Expression and complementation of naphthalene dioxygenase activity in Escherichia coli. in: “Microbial Metabolism and the Carbon Cycle,” S.R. Hagedorn, R.S. Hanson, and D. A. Kunz, ed., Harwood Academic Publishers, New York (1988).Google Scholar
  10. 10.
    T. Isogai, M. Fukagawa, I. Aramori, M. Iwami, H. Kojo, T. Ono, Y. Ueda, M. Kohsaka, and Imanaka, Construction of a 7-aminocephalosporanic acid (7ACA) biosynthetic operon and direct production of 7ACA in Acremonium chrysogenum, Bio/Technology, 9:188–191 (1991).PubMedCrossRefGoogle Scholar
  11. 11.
    J. Fieschko, and T. Ritch, Production of human alpha consensus interferon in recombinant Escherichia coli. Chem. Eng. Commun., 45:229–240 (1986).CrossRefGoogle Scholar
  12. 12.
    K. N. Timmis, F. Rojo, and J. L. Ramos, Prospects for laboratory engineering of bacteria to degrade pollutants in: “Environmental Biotechnology; Reducing Risks from Environmental Chemicals Through Biotechnology,” Ullman, G.S., ed. Plenum Press, New York (1988).Google Scholar
  13. 13.
    A. M. Chakrabarty, Microorganisms having multiple compatible degradative energy generating plasmids and preparation thereof, US patent 4, 259, 444 (1981).Google Scholar
  14. 14.
    J. F. Grindley, M. A. Payton, H. Pole and K. G. Harding, Conversion of Glucose to 2-keto-l-gulonate, an intermediate in l-ascorbate synthesis by recombinant strain of Erwinia citrus, Appl. and Environ. Microbiol. 54:1770–1775 (1988).Google Scholar
  15. 15.
    L. O. Narhi, Y. Stabinski, M. Levitt, L. Miller, R. Sachdev, S. Finley, S. Park, C. Kolvenbach, T. Aurakawa, and M. Zukowski, Enhanced stability of subtilisin by three point mutations, Biotechnol. Appl. Biochem. 13:12–24 (1991).PubMedGoogle Scholar
  16. 16.
    M. Zukowski, Y. Stabinski, L. Narhi, J. Mauck, M. Stowers, and M. Fiske, An engineered subtilisin with improved stability: applications in human diagnostics, in: “Genetics and Biotechnology of Bacilli, Vol. 3,” M.N. Zukowski, A. T. Ganesan, and J. A. Hoch, ed., Academic Press, New York (1990).Google Scholar
  17. 17.
    D. Murdock, M. Thalen, C. Serdar, and B. Ensley, Manipulation of metabolic operons catalyzing the biochemical synthesis of indigo, in press.Google Scholar
  18. 18.
    B. G. Fox, J. G. Borneman, L. P. Wackett, and J. D. Lipscomb, Haloalkene oxidation by the soluble methane monooxygenase from Methylosinus trichosporium OB3b: mechanistic and environmental implications, Biochem., 29:6419–6427, (1990).CrossRefGoogle Scholar
  19. 19.
    J. A. Wells, B. C. Cuningham, T. P. Graycar and D. A. Estell, Recruitment of substrate specificity properties from one enzyme into a related one by protein engineering. Proc. Natl. Acad. Sci. 84:5157–5174 (1987).Google Scholar
  20. 20.
    J. A. Wells, D. B. Bowers, R. R. Bott, T. P. Graycar, and D. A. Estelle, Designing substrate specificity by protein engineering of electrostatic interactions, Proc. Natl. Acad. Sci. 84:1219–1223 (1987).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • Burt D. Ensley
    • 1
  1. 1.Envirogen, Inc.LawrencevilleUSA

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