Insertion of an E. coli lacZ gene in Acetobacter xylinus for the production of cellulose in whey.
A mini-Tn10:lacZ:kan was inserted into a wild-type strain of Acetobacter xylinus by random transposon mutagenesis, generating a lactose-utilising and cellulose-producing mutant strain designated ITz3. Antibiotic selection plate assays and Southern hybridisation revealed that the lacZ gene was inserted once into the chromosome of strain ITz3 and was stably maintained in non-selective medium after more than 60 generations. The modified strain had, on the average, a 28-fold increase in cellulose production and a 160-fold increase in beta-galactosidase activity when grown in lactose medium. beta-Galactosidase activity is present in either lactose or sucrose medium indicating that the gene is constitutively expressed. Cellulose and beta-galactosidase production by the modified strain was also evaluated in pure and enriched whey substrates. Utilisation of lactose in whey substrate by ITz3 reached 17 g l(-1) after 4 days incubation.
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
Evelyn
E
Battad-Bernardo
Sharon L
SL
McCrindle
Iain
I
Couperwhite
Brett A
BA
Neilan
D002482:Cellulose / Q000096:biosynthesis* / Q000378:metabolism; D002611:Cheese / Q000382:microbiology*; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D041822:Gluconacetobacter xylinus / Q000235:genetics* / Q000378:metabolism*; D007763:Lac Operon; D007785:Lactose / Q000378:metabolism; D016296:Mutagenesis; D010957:Plasmids; D001616:beta-Galactosidase / Q000235:genetics
D004251:DNA Transposable Elements; D002482:Cellulose; D001616:beta-Galactosidase; D007785:Lactose