Hexadecane and Tween 80 stimulate lipase production in Burkholderia glumae by different mechanisms.
Burkholderia glumae strain PG1 produces a lipase of biotechnological relevance. Lipase production by this strain and its derivative LU8093, which was obtained through classical strain improvement, was investigated under different conditions. When 10% hexadecane was included in the growth medium, lipolytic activity in both strains could be increased approximately 7-fold after 24 h of growth. Hexadecane also stimulated lipase production in a strain containing the lipase gene fused to the tac promoter, indicating that hexadecane did not affect lipase gene expression at the transcriptional level, which was confirmed using lipA-gfp reporter constructs. Instead, hexadecane appeared to enhance lipase secretion, since the amounts of lipase in the culture supernatant increased in the presence of hexadecane, with a concomitant decrease in the cells, even when protein synthesis was inhibited with chloramphenicol. In the presence of olive oil as a carbon source, nonionic detergents, such as Tween 80, increased extracellular lipase activity twofold. Like hexadecane, Tween 80 appeared to stimulate lipase secretion, although in a more disruptive manner, since other, normally nonsecreted proteins were found in the culture supernatant. Additionally, like olive oil, Tween 80 was found to induce lipase gene expression in strain PG1 in medium containing sucrose as a carbon source but not in glucose-containing medium, suggesting that lipase gene expression is prone to catabolite repression. In contrast, lipase production in the lipase-overproducing strain LU8093 was independent of the presence of an inducer and was not inhibited by glucose. In conclusion, hexadecane and Tween 80 enhance lipase production in B. glumae, and they act via different mechanisms.
Boekema
Bouke K H L
BK
Department of Molecular Microbiology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Beselin
Anke
A
Breuer
Michael
M
Hauer
Bernhard
B
Koster
Margot
M
Rosenau
Frank
F
Jaeger
Karl-Erich
KE
Tommassen
Jan
J
D000473:Alkanes / Q000494:pharmacology*; D015153:Blotting, Western; D019117:Burkholderia / Q000201:enzymology*; D017931:DNA Primers; D018274:Electroporation; D005453:Fluorescence; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D011136:Polysorbates / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D013501:Surface-Active Agents / Q000494:pharmacology*
D000473:Alkanes; D017931:DNA Primers; D011136:Polysorbates; D013501:Surface-Active Agents; D008049:Lipase; C007932:n-hexadecane