Team:TU Munich/Project/Limonene


Revision as of 21:14, 23 September 2012 by Fabian Froehlich (Talk | contribs)



Responsible: Andrea Richter and Lara Kuntz

Producing the flavoring substance limonene in our beer might result in a fresh, lemon-like taste on the one hand. On the other hand, we might have health beneficial effects like preventive activity against cancer, dissolution of gallstones and relief of heartburn.

  • bring this in the form of a motivational statement on why we want to have limonene in our beer.

Background and principles

Limonene is a cyclic terpene and a major constituent of several citrus oils (orange, lemon, mandarin, lime and grapefruit). It is a chiral molecule; citrus fruits contain the (R)-enantiomer. The (R)-enantiomer smells like oranges, while the (S)-enantionmer has a piney, turpentine-like odor. D-Limonene is used as a component of flavorings and fragrances.


Limonene is formed from geranyl pyrophosphat by limonene synthase. (R)-limonene synthase 1 consists of 606 aminoacids (sequence shown below, EC= (R)-limonene synthase needs magnesium or manganese as cofactor and catalyzes the following reaction: Geranyl pyrophosphate = (+)-(4R)-limonene + diphosphate.

  • state that geranyl pyrophosphate is naturally available in the yeast metabolism + reference

TUM12 ReactionLimoneneSynthase.jpg

The molecular and physiological effects of limonene

It has been shown to inhibit rat mammary and other tumor development (Tsuda et al. 2004). Being an excellent solvent of cholesterol, d-limonene also has been used clinically to dissolve cholesterol-containing gallstones. Because of its gastric acid neutralizing effect and its support of normal peristalsis, it has also been used for relief of heartburn (Sun 2007).

  • elaborate a bit more


Limonene can be produced by (R)-limonene synthase.

  • state how we want to indtroduce this biobrick in S. cerevisiae


Gel Picture of finished construct

SDS Page of Limonene Synthase

GCMS spectrum of Limonene

Toxicity Assay


Herrero O, Ramón D, Orejas M: Engineering the Saccharomyces cerevisiae isoprenoid pathway for de novo production of aromatic monoterpenes in wine. Metab Eng 2008 10:78-86.

Lücker J, Tamer M, Schwab W, Verstappen F, Van der Plas L, Bouwmeester H and Verhoeven H: Monoterpene biosynthesis in lemon (Citrus limon) cDNA isolation and functional analysis of four monoterpene synthases. Eur J Biochem 2002 269:3160-3171.

Sun J: D-Limonene: Safety and Clinical Applications. Altern Med Rev 2007 12(3):259-264.

Tsuda H, Ohshim Y, Nomoto H, Fujita K, Matsuda E, Iigo M, Takasuka N, Moore M: Cancer Prevention by Natural Compounds. Drug Metab Pharmacokin 2004 19(4):245-263.