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. 1990 Apr;56(4):919–923. doi: 10.1128/aem.56.4.919-923.1990

Construction of lactose-utilizing Xanthomonas campestris and production of xanthan gum from whey.

J F Fu 1, Y H Tseng 1
PMCID: PMC184322  PMID: 2111116

Abstract

Xanthomonas campestris pv. campestris possesses a low level of beta-galactosidase and therefore is not able to grow and produce significant amounts of xanthan gum in a medium containing lactose as the sole carbon source. In this study, a beta-galactosidase expression plasmid was constructed by ligating an X. campestris phage phi LO promoter with pKM005, a ColE1 replicon containing Escherichia coli lacZY genes and the lpp ribosome-binding site. It was then inserted into an IncP1 broad-host-range plasmid, pLT, and subsequently transferred by conjugation to X. campestris 17, where it was stably maintained. The lacZ gene under the control of the phage promoter was expressed at a high level, enabling the cells to grow in a medium containing lactose. Production of xanthan gum in lactose or diluted whey by the engineered strain was evaluated, and it was found to produce as much xanthan gum in these substrates as the cells did in a medium containing glucose.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cornelis G., Ghosal D., Saedler H. Tn951: a new transposon carrying a lactose operon. Mol Gen Genet. 1978 Apr 6;160(2):215–224. doi: 10.1007/BF00267484. [DOI] [PubMed] [Google Scholar]
  4. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frank J. F., Somkuti G. A. General Properties of Beta-Galactosidase of Xanthomonas campestris. Appl Environ Microbiol. 1979 Sep;38(3):554–556. doi: 10.1128/aem.38.3.554-556.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  7. Jansson P. E., Kenne L., Lindberg B. Structure of extracellular polysaccharide from Xanthomonas campestris. Carbohydr Res. 1975 Dec;45:275–282. doi: 10.1016/s0008-6215(00)85885-1. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Melton L. D., Mindt L., Rees D. A., Sanderson G. R. Covalent structure of the extracellular polysaccharide from Xanthomonas campestris: evidence from partial hydrolysis studies. Carbohydr Res. 1976 Feb;46(2):245–257. doi: 10.1016/s0008-6215(00)84296-2. [DOI] [PubMed] [Google Scholar]
  10. Schwartz R. D., Bodie E. A. Production of High-Viscosity Whey Broths by a Lactose-Utilizing Xanthomonas campestris Strain. Appl Environ Microbiol. 1985 Dec;50(6):1483–1485. doi: 10.1128/aem.50.6.1483-1485.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Walsh P. M., Haas M. J., Somkuti G. A. Genetic Construction of Lactose-Utilizing Xanthomonas campestris. Appl Environ Microbiol. 1984 Feb;47(2):253–257. doi: 10.1128/aem.47.2.253-257.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Yang B. Y., Tsai H. F., Tseng Y. H. Broad host range cosmid pLAFR1 and non-mucoid mutant XCP20 provide a suitable vector-host system for cloning genes in Xanthomonas campestris pv. campestris. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi. 1988 Feb;21(1):40–49. [PubMed] [Google Scholar]

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