1887

Abstract

-Amino acids, important intermediates in the production of semisynthetic penicillins and cephalosporins, are currently prepared from the corresponding hydantoins using bacterial biomass containing two enzymes, hydantoinase and carbamylase. These enzymes convert the hydantoins first into carbamyl derivatives and then into the corresponding -amino acids. In an attempt to select more efficient biocatalysts, the hydantoinase and carbamylase genes from (formerly ) were cloned in The genes were assembled to give two operon-type structures one having the carbamylase gene preceding the hydantoinase gene and the other with the carbamylase gene following the hydantoinase gene. The recombinant strains stably and constitutively produced the two enzymes and efficiently converted the corresponding hydantoins into -hydroxyphenyl-glycine and phenylglycine. The order of the genes within the operon and the growth temperature of the strains turned out to be important for both enzyme and -amino acid production. The configuration with the carbamylase gene preceding the hydantoinase gene was the most efficient one when the biomass was grown at 25°C rather than 37°C. This biomass produced -amino acid twice as efficiently as the industrial strain of The efficiency was found to be correlated with the level of carbamylase produced, indicating that the concentration of this enzyme is the rate-limiting factor in -amino acid production under the conditions used on an industrial scale.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-144-4-947
1998-04-01
2021-05-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/144/4/mic-144-4-0947.html?itemId=/content/journal/micro/10.1099/00221287-144-4-947&mimeType=html&fmt=ahah

References

  1. Ambler R. P., Auffret A. D., Clarke P. H. (1987); The amino acid sequence of the aliphatic amidase from Pseudomonas aeruginosa.. Febs Letters 215:(2),285–290 [CrossRef]
    [Google Scholar]
  2. Buckholz R. G., Cooper T. G. (1991); The allantoinase {DALI) gene of Saccharomyces cerevisiae.. Yeast 7:(9),913–923 [CrossRef]
    [Google Scholar]
  3. Dagert M., Ehlrich S. D. (1979); Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells.. Gene 6:(1),23–28 [CrossRef]
    [Google Scholar]
  4. Dower W. G., Miller J. F., Ragsdale C. W. (1988); High efficiency transformation of E. coli by high voltage electroporation.. Nucleic Acids Research 16:(13),6127–6145 [CrossRef]
    [Google Scholar]
  5. Grifantini R., Pratesi P., Galli G., Grandi G. (1996); Topological mapping of cysteine residues of N-carbamyl-D-amino-acid amido- hydrolase and their role in enzymic activity.. Journal of Biological Chemistry 271:(16),9326–9331 [CrossRef]
    [Google Scholar]
  6. Kim G.-J., Kim H.-S. (1995); Optimization of the enzymic synthesis of D-p-hydroxyphenylglycine from DL-monosubstituted hydantoin using D-hydantoinase and N-carbamylase.. Enzyme Microb Technol 17:(1),63–65 [CrossRef]
    [Google Scholar]
  7. Labigne A., Cussac V., Courcoux P. (1991); Shuttle cloning and nucleotide sequence of Helicobacter pylori genes responsible for urease activity.. Journal of Bacteriology 173:(6),1920–1931 [CrossRef]
    [Google Scholar]
  8. Laemmli U. K. (1970); Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:(5259),680–685 [CrossRef]
    [Google Scholar]
  9. LaPointe G., Viau S., Leblanc D., Robert N., Morin, A. (1994); Cloning, sequencing and expression in Escherichia coli of the d- hydantoinase gene from Pseudomonas putida and distribution of homologous genes in other micro-organisms. Appl Environ Microbiol 60:888–895
    [Google Scholar]
  10. Lee, S.-G., Lee, D.-C., Sung, M.-H., Kim H.-S. (1994); Isolation of thermostable D-hydantoinase-producing thermophilic Bacillus sp.. SD-1. Biotechnol Lett 16:(5),461–166 [CrossRef]
    [Google Scholar]
  11. Meyer P., Runser S. (1993); Efficient production of the industrial biocatalists hydantoinase and N-carbamyl amino acid amido- hydrolase.. Fems Microbiology Letters 109:(1),67–74 [CrossRef]
    [Google Scholar]
  12. Ogawa J., Shimizu S., Yamada H. (1994a); N-Carbamoyl-D- amino acid amidohydrolase from Comamonas sp. E222c, purification and characterization.. European Journal of Biochemistry 212:(3),685–691 [CrossRef]
    [Google Scholar]
  13. Ogawa J., Chung M. C. M., Hida S., Yamada H., Shimizu S. (1994b); Thermostable N-carbamoyl-D-amino acid amidohydrolase: screening, purification and characterization.. J Biotechnol 38:11–19 [CrossRef]
    [Google Scholar]
  14. Olivieri R., Fascetti E., Angelini L., Degen L. (1981); Microbial transformation of racemic hydantoins to D-amino acids.. Biotechnology and Bioengineering 23:(10),2173–2183 [CrossRef]
    [Google Scholar]
  15. Runser S., Ohleyer E. (1990); Properties of the hydantoinase from Agrobacterium sp. IP 1-671.. Biotechnology Letters 12:(4),259–264 [CrossRef]
    [Google Scholar]
  16. Runser S., Chinski N., Ohleyer E. (1990); D-p-Hydroxy- phenylglycine production from DL-5-p-hydroxyphenylhydantoin by Agrobacterium sp.. Appl Microbiol Biotechnol 33:(4),382–388 [CrossRef]
    [Google Scholar]
  17. Sambrook J., Fritsh E. F., Maniatis T. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. (1977); DNA sequencing with chain terminating inhibitors.. Proc Natl Acad Sci USA 74:(12),5463–5467 [CrossRef]
    [Google Scholar]
  19. Soubrier F., Levy-Scill S., Mayaux J. F., Petra D., Arnaud A., Couzet J. (1992); Cloning and primary structure of the wide- spectrum amidase from Brevibacterium sp. R312: high homology to the amiE product from Pseudomonas aeruginosa.. Gene 16:(1),99–104 [CrossRef]
    [Google Scholar]
  20. Stark G. R., Smyth D. G. (1963); The use of cyanate for the determination of NH2-terminal residues in proteins.. Journal of Biological Chemistry 238:(1),214–226 [CrossRef]
    [Google Scholar]
  21. Suzuki T., Igarashi K., Hase K., Tsujimura K. (1973); Optical rotary dispersion and circular dichroism of amino acid hydantoins.. Agric Biol Chem 37:(2),411–416 [CrossRef]
    [Google Scholar]
  22. Takahashi S., Ohashi T., Kii Y., Kumagai H., Yamada H. (1979); Microbial transformation of hydantoins to N-carbamyl- D-amino acids.. Journal of Fermentation Technology 57:328–332
    [Google Scholar]
  23. Velati Bellini A., Galli G., Fascetti E., Frascotti G., Branduzzi P., Lucchese G., Grandi G. (1991); Production processes of recombinant IL-1β from Bacillus subtilis: comparison between intracellular and exocellular expression.. Journal of Biotechnology 18:(3),177–192 [CrossRef]
    [Google Scholar]
  24. Watabe K., Ishikawa T., Mukohara Y., Nakamura H. (1992); Cloning and sequencing of the genes involved in the conversion of 5-substituted hydantoins to the corresponding L-amino acids from the native plasmid of Pseudomonas sp. strain NS671. Journal of Bacteriology 174962–969
    [Google Scholar]
  25. Weatherburn M. W. (1967); Phenol—hypochlorite reaction for determination of ammonia.. Analytical Chemistry 207:285–290
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-144-4-947
Loading
/content/journal/micro/10.1099/00221287-144-4-947
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error