This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Craft, D. L. Articles by Wilson, C. R. Search for Related Content PubMed PubMed Citation Articles by Craft, D. L. Articles by Wilson, C. R. Agricola Articles by Craft, D. L. Articles by Wilson, C. R.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, October 2003, p. 5983-5991, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5983-5991.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of the CYP52 Family of Candida tropicalis ATCC 20336, Important for the Conversion of Fatty Acids and Alkanes to ,-Dicarboxylic Acids

David L. Craft,^* Krishna M. Madduri, Mark Eshoo, and C. Ron Wilson

Biotechnology Group, Cognis Corporation, Cincinnati, Ohio 45232

Received 20 March 2003/ Accepted 25 July 2003

Candida tropicalis ATCC 20336 excretes ,-dicarboxylic acids as a by-product when cultured on n-alkanes or fatty acids as the carbon source. Previously, a ß-oxidation-blocked derivative of ATCC 20336 was constructed which showed a dramatic increase in the production of dicarboxylic acids. This paper describes the next steps in strain improvement, which were directed toward the isolation and characterization of genes encoding the -hydroxylase enzymes catalyzing the first step in the -oxidation pathway. Cytochrome P450 monooxygenase (CYP) and the accompanying NADPH cytochrome P450 reductase (NCP) constitute the hydroxylase complex responsible for the first and rate-limiting step of -oxidation of n-alkanes and fatty acids. 10 members of the alkane-inducible P450 gene family (CYP52) of C. tropicalis ATCC20336 as well as the accompanying NCP were cloned and sequenced. The 10 CYP genes represent four unique genes with their putative alleles and two unique genes for which no allelic variant was identified. Of the 10 genes, CYP52A13 and CYP52A14 showed the highest levels of mRNA induction, as determined by quantitative competitive reverse transcription-PCR during fermentation with pure oleic fatty acid (27-fold increase), pure octadecane (32-fold increase), and a mixed fatty acid feed, Emersol 267 (54-fold increase). The allelic pair CYP52A17 and CYP52A18 was also induced under all three conditions but to a lesser extent. Moderate induction of CYP52A12 was observed. These results identify the CYP52 and NCP genes as being involved in ,-dicarboxylic acid production by C. tropicalis and provide the foundation for biocatalyst improvement.

Present address: Dow AgroSciences LLC, Indianapolis, IN 46268.

Present address: Buck Institute, Novato, CA 94945.

This article has been cited by other articles:

• Funhoff, E. G., Bauer, U., Garcia-Rubio, I., Witholt, B., van Beilen, J. B. (2006). CYP153A6, a Soluble P450 Oxygenase Catalyzing Terminal-Alkane Hydroxylation.. J. Bacteriol. 188: 5220-5227 [Abstract] [Full Text]  
• Ko, B. S., Kim, J., Kim, J. H. (2006). Production of Xylitol from D-Xylose by a Xylitol Dehydrogenase Gene-Disrupted Mutant of Candida tropicalis.. Appl. Environ. Microbiol. 72: 4207-4213 [Abstract] [Full Text]  
• Eirich, L. D., Craft, D. L., Steinberg, L., Asif, A., Eschenfeldt, W. H., Stols, L., Donnelly, M. I., Wilson, C. R. (2004). Cloning and Characterization of Three Fatty Alcohol Oxidase Genes from Candida tropicalis Strain ATCC 20336. Appl. Environ. Microbiol. 70: 4872-4879 [Abstract] [Full Text]