caa a22 4500 465824234 CHVBK 20180323112144.0 cr unu---uuuuu 170327e19900701xx s 000 0 eng 10.1007/BF00176656 doi (NATIONALLICENCE)springer-10.1007/BF00176656 Lactose- and galactose-utilizing strains of poly(hydroxyalkanoic acid)-accumulating Alcaligenes eutrophus and Pseudomonas saccharophila obtained by recombinant DNA technology [Elektronische Daten] [Andreas Pries, Alexander Steinbüchel, Hans Schlegel] Summary: Transposon Tn 951-encoded β-galactosidase was expressed in Pseudomonas saccharophila and enabled this bacterium to grow on lactose as sole carbon source. In contrast, β-galactosidase was not expressed in Alcaligenes eutrophus even if the lacZ gene of Tn 951 was separated from the lacI gene. However, β-galactosidase was expressed in A. eutrophus, if a DNA fragment, which was suspected to harbour the promoter of the A. eutrophus poly(3-hydroxybutyric acid)-synthetic genes, was ligated to the promoter probe vector pMC1403, which employs lac Z, Y as reporter genes. Plasmid pPL76, which harboured one of the promoter-lac fusions, enabled A. eutrophus not only to express β-galactosidase but also to grow slowly on lactose (doubling time = 25-30 h). Subsequently, the promoter-lac fusion was ligated to Tn5 in pSUP5011 and was inserted into the genome of A. eutrophus H16 and of the glucose-utilizing mutant H16-G+1 by applying the suicide plasmid technique. Two recombinant strains, H16-cPL and H16-G+1-cPL, which grow with a doubling time of 16-23 h on lactose, were investigated in detail. The cells only utilized the glucose residue of lactose as a carbon source for grouth and excreted galactose into the medium. Only after the Escherichia coli gal operon had been cloned in vector pVK101 and had been mobilized to H16-cPL or H16-G+1-cPL, was lactose completely utilized; no galactose was detected in the medium and the growth yields increased twofold. Depending on the orientation of the gal operon in pVK101, the expression of galactokinase seems to be dependent either on the promoter of aminoglycoside phosphotransferase gene (kan) or on the promoter of the tetR gene. Springer-Verlag, 1990 Pries Andreas Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut Steinbüchel Alexander Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut Schlegel Hans Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut Applied Microbiology and Biotechnology Springer-Verlag 33/4(1990-07-01), 410-417 0175-7598 33:4<410 1990 33 253 https://doi.org/10.1007/BF00176656 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00176656 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Pries Andreas Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut NATIONALLICENCE 700 1- Steinbüchel Alexander Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut NATIONALLICENCE 700 1- Schlegel Hans Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 33/4(1990-07-01), 410-417 0175-7598 33:4<410 1990 33 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer