Applied Microbiology and Biotechnology

, Volume 93, Issue 1, pp 159–168 | Cite as

Synthesis of green note aroma compounds by biotransformation of fatty acids using yeast cells coexpressing lipoxygenase and hydroperoxide lyase

  • Markus Buchhaupt
  • Jan Christopher Guder
  • Maria Magdalena Walburga Etschmann
  • Jens Schrader
Biotechnological products and process engineering

Abstract

Green notes are substances that characterize the aroma of freshly cut grass, cucumbers, green apples, and foliage. In plants, they are synthesized by conversion of linolenic or linoleic acid via the enzymes lipoxygenase (LOX) and hydroperoxide lyase (HPL) to short-chained aldehydes. Current processes for production of natural green notes rely on plant homogenates as enzyme sources but are limited by low enzyme concentration and low specificity. In an alternative approach, soybean LOX2 and watermelon HPL were overexpressed in Saccharomyces cerevisiae. After optimization of the expression constructs, a yeast strain coexpressing LOX and HPL was applied in whole cell biotransformation experiments. Whereas addition of linolenic acid to growing cultures of this strain yielded no products, we were able to identify high green note concentrations when resting cells were used. The primary biotransformation product was 3(Z)-hexenal, a small amount of which isomerized to 2(E)-hexenal. Furthermore, both aldehydes were reduced to the corresponding green note alcohols by endogenous yeast alcohol dehydrogenase to some extent. As the cosolvent ethanol was the source of reducing equivalents for green note alcohol formation, the hexenal/hexenol ratio could be influenced by the use of alternative cosolvents. Further investigations to identify the underlying mechanism of the rather low biocatalyst stability revealed a high toxicity of linolenic acid to yeast cells. The whole cell catalyst containing LOX and HPL enzyme activity described here can be a promising approach towards a highly efficient microbial green note synthesis process.

Keywords

Green notes Saccharomyces cerevisiae Lipoxygenase Hydroperoxide lyase Fatty acid Biotransformation 

Notes

Acknowledgments

The authors acknowledge Wilfried Schwab for providing plasmids containing LOX and HPL cDNAs. Financial support from Arbeitsgemeinschaft Industrieller Fördervereinigungen e.V. under the project 15088N is gratefully acknowledged.

Conflicts of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Markus Buchhaupt
    • 1
  • Jan Christopher Guder
    • 1
  • Maria Magdalena Walburga Etschmann
    • 1
  • Jens Schrader
    • 1
  1. 1.Biochemical EngineeringDECHEMA e.V., Karl Winnacker-InstitutFrankfurtGermany

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