Three distinct quinoprotein alcohol dehydrogenases are expressed when Pseudomonas putida is grown on different alcohols.
A bacterial strain that can utilize several kinds of alcohols as its sole carbon and energy sources was isolated from soil and tentatively identified as Pseudomonas putida HK5. Three distinct dye-linked alcohol dehydrogenases (ADHs), each of which contained the prosthetic group pyrroloquinoline quinone (PQQ), were formed in the soluble fractions of this strain grown on different alcohols. ADH I was formed most abundantly in the cells grown on ethanol and was similar to the quinoprotein ADH reported for P. putida (H. Görisch and M. Rupp, Antonie Leeuwenhoek 56:35-45, 1989) except for its isoelectric point. The other two ADHs, ADH IIB and ADH IIG, were formed separately in the cells grown on 1-butanol and 1,2-propanediol, respectively. Both of these enzymes contained heme c in addition to PQQ and functioned as quinohemoprotein dehydrogenases. Potassium ferricyanide was an available electron acceptor for ADHs IIB and IIG but not for ADH I. The molecular weights were estimated to be 69,000 for ADH IIB and 72,000 for ADH IIG, and both enzymes were shown to be monomers. Antibodies raised against each of the purified ADHs could distinguish the ADHs from one another. Immunoblot analysis showed that ADH I was detected in cells grown on each alcohol tested, but ethanol was the most effective inducer. ADH IIB was formed in the cells grown on alcohols of medium chain length and also on 1,3-butanediol. Induction of ADH IIG was restricted to 1,2-propanediol or glycerol, of which the former alcohol was more effective. These results from immunoblot analysis correlated well with the substrate specificities of the respective enzymes. Thus, three distinct quinoprotein ADHs were shown to be synthesized by a single bacterium under different growth conditions.
Toyama
H
H
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Japan.
Fujii
A
A
Matsushita
K
K
Shinagawa
E
E
Ameyama
M
M
Adachi
O
O
D020001:1-Butanol; D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism*; D000440:Butanols / Q000378:metabolism; D003429:Cross Reactions; D000431:Ethanol / Q000378:metabolism; D015870:Gene Expression; D015151:Immunoblotting; D045542:PQQ Cofactor; D019946:Propylene Glycol; D011409:Propylene Glycols / Q000378:metabolism; D016958:Pseudomonas putida / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000254:growth & development; D015363:Quinolones / Q000302:isolation & purification*; D013045:Species Specificity; D013053:Spectrophotometry; D013379:Substrate Specificity
D000438:Alcohols; D000440:Butanols; D011409:Propylene Glycols; D015363:Quinolones; D000431:Ethanol; D019946:Propylene Glycol; D045542:PQQ Cofactor; D020001:1-Butanol; D000426:Alcohol Dehydrogenase