Team:TU Munich/Project/Limonene

From 2012.igem.org

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==Idea==
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Limonene can be produced by (R)-limonene synthase.
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* state how we want to indtroduce this biobrick in ''S. cerevisiae''
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==Results==
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*Landmann C, Fink B, Festner M, Dregus M, Engel KH, Schwab W '''(2007)''' Cloning and functional characterization of three terpene synthases from lavender (Lavandula angustifolia). Arch Biochem Biophys 467:417–429.
*Landmann C, Fink B, Festner M, Dregus M, Engel KH, Schwab W '''(2007)''' Cloning and functional characterization of three terpene synthases from lavender (Lavandula angustifolia). Arch Biochem Biophys 467:417–429.
*William DC, McGarvey DJ, Katahira EJ, Croteau R '''(1998)''' Truncation of Limonene Synthase Preprotein Provides a Fully Active ‘Pseudomature’ Form of This Monoterpene Cyclase and Reveals the Function of the Amino-Terminal Arginine Pair. Biochem 37:12213-12220.
*William DC, McGarvey DJ, Katahira EJ, Croteau R '''(1998)''' Truncation of Limonene Synthase Preprotein Provides a Fully Active ‘Pseudomature’ Form of This Monoterpene Cyclase and Reveals the Function of the Amino-Terminal Arginine Pair. Biochem 37:12213-12220.
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== older blocks ==
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- please integrate the informations into the other blocks
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 +
===Idea===
 +
----
 +
<div>
 +
Limonene can be produced by (R)-limonene synthase.
 +
* state how we want to indtroduce this biobrick in ''S. cerevisiae''
 +
</div>

Revision as of 18:49, 24 September 2012


Contents

Limonene


Responsible: Andrea Richter and Lara Kuntz

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. 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).

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 beneficial effects on health such as preventive activity against cancer, dissolution of gallstones and relief of heartburn.


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.

Biosynthesis

Limonene is formed from geranyl pyrophosphat by limonene synthase. (R)-limonene synthase 1 consists of 606 aminoacids (sequence shown below, EC=4.2.3.20). (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

Results


Gel Picture of finished construct


SDS Page of Limonene Synthase


GCMS spectrum of Limonene


Toxicity Assay

Evaluation of the Toxicity Assay for Limonene.

References


  • Herrero O, Ramón D, Orejas M (2008) Engineering the Saccharomyces cerevisiae isoprenoid pathway for de novo production of aromatic monoterpenes in wine. Metab Eng 10:78-86.
  • Lücker J, Tamer M, Schwab W, Verstappen F, Van der Plas L, Bouwmeester H, Verhoeven H (2002) Monoterpene biosynthesis in lemon (Citrus limon) cDNA isolation and functional analysis of four monoterpene synthases. Eur J Biochem 269:3160-3171.
  • Sun J (2007) D-Limonene: Safety and Clinical Applications. Altern Med Rev 12(3):259-264.
  • Tsuda H, Ohshim Y, Nomoto H, Fujita K, Matsuda E, Iigo M, Takasuka N, Moore M (2004) Cancer Prevention by Natural Compounds. Drug Metab Pharmacokin 19(4):245-263.
  • Landmann C, Fink B, Festner M, Dregus M, Engel KH, Schwab W (2007) Cloning and functional characterization of three terpene synthases from lavender (Lavandula angustifolia). Arch Biochem Biophys 467:417–429.
  • William DC, McGarvey DJ, Katahira EJ, Croteau R (1998) Truncation of Limonene Synthase Preprotein Provides a Fully Active ‘Pseudomature’ Form of This Monoterpene Cyclase and Reveals the Function of the Amino-Terminal Arginine Pair. Biochem 37:12213-12220.


older blocks

- please integrate the informations into the other blocks

Idea


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

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