Two novel metal-independent long-chain alkyl alcohol dehydrogenases from Geobacillus thermodenitrificans NG80-2.
Two alkyl alcohol dehydrogenase (ADH) genes from the long-chain alkane-degrading strain Geobacillus thermodenitrificans NG80-2 were characterized in vitro. ADH1 and ADH2 were prepared heterologously in Escherichia coli as a homooctameric and a homodimeric protein, respectively. Both ADHs can oxidize a broad range of alkyl alcohols up to at least C(30), as well as 1,3-propanediol and acetaldehyde. ADH1 also oxidizes glycerol, and ADH2 oxidizes isopropyl alcohol, isoamylol, acetone, octanal and decanal. The best substrate is ethanol for ADH1 and 1-octanol for ADH2. For both ADHs, the optimum assay condition is at 60 degrees C and pH 8.0, and both NAD and NADP can be used as the cofactor. Sequence analysis reveals that ADH1 and ADH2 belong to the Fe-containing/activated long-chain ADHs. However, the two enzymes contain neither Fe nor other metals, and Fe is not required for the activity, suggesting a new type of ADH. The ADHs characterized here are potentially useful in crude oil bioremediation and other bioconversion processes.
Liu
Xueqian
X
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Dong
Yanpeng
Y
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Zhang
Jing
J
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Zhang
Aixiang
A
Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
Wang
Lei
L
TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.|The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, PR China.|The Engineering and Research Center for Microbial Functional Genomics and Detection Technology, Ministry of Education, PR China.|Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.
Feng
Lu
L
The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, PR China.|The Engineering and Research Center for Microbial Functional Genomics and Detection Technology, Ministry of Education, PR China.|Tianjin Research Center for Functional Genomics and Biochip, Tianjin 300457, PR China.|Tianjin Key Laboratory of Microbial Functional Genomics, Tianjin 300457, PR China.|TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin 300457, PR China.
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism; D000595:Amino Acid Sequence; D001406:Bacillaceae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D001673:Biodegradation, Environmental; D002614:Chelating Agents / Q000494:pharmacology; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004492:Edetic Acid / Q000494:pharmacology; D007501:Iron / Q000378:metabolism; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D017421:Sequence Analysis; D013379:Substrate Specificity
D000438:Alcohols; D002614:Chelating Agents; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D009243:NAD; D009249:NADP; D004492:Edetic Acid; D007501:Iron; D000429:Alcohol Oxidoreductases; C065402:long-chain-alcohol dehydrogenase