Differences in substrate specificities of five bacterial wax ester synthases

Brett M. Barney, Bradley D. Wahlen, Emma Lee Garner, Jiashi Wei, Lance C. Seefeldt

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

Abstract

Wax esters are produced in certain bacteria as a potential carbon and energy storage compound. The final enzyme in the biosyn- thetic pathway responsible for wax ester production is the bifunctional wax ester synthase/acyl-coenzyme A (acyl-CoA):diacyl- glycerol acyltransferase (WS/DGAT), which utilizes a range of fatty alcohols and fatty acyl-CoAs to synthesize the corresponding wax ester. We report here the isolation and substrate range characterization for five WS/DGAT enzymes from four different bacteria: Marinobacter aquaeolei VT8, Acinetobacter baylyi, Rhodococcus jostii RHA1, and Psychrobacter cryohalolentis K5. The results from kinetic studies of isolated enzymes reveal a differential activity based on the order of substrate addition and reveal subtle differences between the substrate selectivity of the different enzymes. These in vitro results are compared to the wax ester and triacylglyceride product profiles obtained from each organism grown under neutral lipid accumulating conditions, provid- ing potential insights into the role that the WS/DGAT enzyme plays in determining the final wax ester products that are produced under conditions of nutrient stress in each of these bacteria. Further, the analysis revealed that one enzyme in particular from M. aquaeolei VT8 showed the greatest potential for future study based on rapid purification and significantly higher activ- ity than was found for the other isolated WS/DGAT enzymes. The results provide a framework to test prospective differences between these enzymes for potential biotechnological applications such as high-value petrochemicals and biofuel production.

Original languageEnglish (US)
Pages (from-to)5734-5745
Number of pages12
JournalApplied and environmental microbiology
Volume78
Issue number16
DOIs
StatePublished - Aug 2012

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