Abstract
The entire Escherichia coli lactose operon was inserted into an E. coli/Corynebacterium glutamicum shuttle vector and introduced into the gram-positive host organism C. glutamicum R163. Recombinant C. glutamicum strains carrying the lac genes downstream of an efficient promoter displayed rapid growth with lactose as the sole source of carbon. Two prerequisites were necessary to obtain good growth of C. glutamicum R 163 on lactose: presence of the lacY gene in addition to lacZ and an appropriatc promoter for efficient transcription in C. glutamicum. The galactose moiety of lactose was not utilized but accumulated in the culture broth. C. glutamicum strains carrying only the lacZ (β-galactosidase) gene but not lacY (lactose permease) were not able to grow in lactose minimal medium.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology. Wiley and Sons, New York
Hennecke H, Günther I, Binder F (1982) A novel cloning vector for the direct selection of recombinant DNA in Escherichia coli. Gene 19: 231–234
Huber RE, Kurz G, Wallenfels K (1976) A quantitation of the factors which affect the hydrolase and transgalactosylase activities of β-galactosidase (E. coli) on lactose. Biochemistry 15: 1994–2001
Kinoshita S, Nakayama K (1978) Amino acids. In: Rose AH (ed) Primary products of metabolism. Academic Press, London, pp 209–261
Liebl W, Schein B (1990) Isolation of restriction deficient mutants of Corynebacterium glutamicum. In: Behrens D, Krämer P (eds) Dechema Biotechnology Conferences, vol. 4 Verlag Chemic, Weinheim, pp 323–327
Liebl W, Klamer R, Schleifer KH (1989a) Requirement of chelating compounds for the growth of Corynebacterium glutamicum in synthetic media. Appl Microbiol Biotechnol 32: 205–210
Liebl W, Bayerl A, Schein B, Stillner U, Schleifer KH (1989b) High efficiency electroporation of intact Corynebacterium glutamicum cells. FEMS Microbiol Lett 65: 299–304
Miller JH (1972) Experiments in molecular genetics Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Nakamaye KI, Eckstein F (1986) Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis Nucleic Acids Res 14: 9679–9698
Padan E, Arbel T, Rimon A, Shira AB, Cohen A (1983) Biosynthesis of the lactose permease in Escherichia coli minicells and effect of carrier amplification on cell physiology. J Biol Chem 258: 5666–5673
Pries A, Steinbüchel A, Schlegel HG (1990) Lactose- and galactose-utilizing strains of poly(hydroxyalkanoic acid)-accumulating Alcaligenes eutrophus and Pseudomonas saccharophila obtained by recombinant DNA technology. Appl Microbiol Biotechnol 33: 410–417
Schäfer A, Kalinowski J, Simon R, Seep-Feldhaus AH, Pühler A (1990) High-frequency conjugal transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria. J Bacteriol 172: 1663–1666
Shapira SK, Chou J, Richaud FR, Casadaban MJ (1983) New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequenzes encoding an enzymatically active carboxy-terminal portion of β-galactosidase. Gene 25: 71–82
Viera J, Messing J (1982) The pUC plasmid, an M13mp/-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19: 259–268
Yoshihama M, Higashiro K, Rao EA, Akedo M, Shanabruch WG, Follettie MT, Waler GC, Sinskey AJ (1985) Cloning vector system for Corynebacterium glutamicum. Bacteriol 162: 591–597
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Brabetz, W., Liebl, W. & Schleifer, KH. Studies on the utilization of lactose by Corynebacterium glutamicum, bearing the lactose operon of Escherichia coli . Arch. Microbiol. 155, 607–612 (1991). https://doi.org/10.1007/BF00245357
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00245357