Limonene Synthase: Detailed information on the kinetic values obtained from the literature

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Kinetic Parameters

Turnover numbers (Kcat)

Parameter Value Units Description References
kcatforward 3.9x10-5 s-1 Limonene synthase expressed at 37°C, 40µM GPP, Citrus sinensis, product formation 1.17x105 µM/sec 
An excerpt from Olsen 2011 Thesis
[1]
kcatforward 0.13 s-1 Limonene synthase expressed at 20°C, 200µM GPP, Citrus sinensis, pH 7.5

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
kcatforward 0.04 s-1 Limonene synthase expressed at 37°C, 200µM GPP, Citrus sinensis, pH 7.5

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
Kcat 0.3 s-1 An excerpt from Alonso 1992 [3]
Kcat 0.020 s-1 An excerpt from Williams 1998, Result section [4]
Kcat 0.024 s-1 Below is an excerpt from Williams 1998, but data source is from Ogura 1997, describing data from wildtype E.coli

An excerpt from Williams 1998, Result section

[5]
Kcat 0.082 s-1 Kcat value for CsTPS1, Cannabis sativa, pH6.5, 40°C, enzyme assay

An excerpt from Günnewich 2006

[6]

Michaelis-Menten constant (Km)

Parameter Value Units Description References
KmGPP 130 µM An excerpt from Entova 2013 Thesis, Chapter 3 [2]
Km 0.7 µM An excerpt from Lucker 2002, Materials and Methods section [7]
KmGPP 1.25 µM An excerpt from Adam 1996, Materials and Methods section [8]
KmGPP 6.8 µM Km value for CsTPS1, pH6.5, 40°C, Cannabis sativa enzyme assay

An excerpt from Günnewich 2006

[6]
KmGPP 8.6 µM Km value for E.coli Q53, from Williams 1998

An excerpt from Günnewich 2006

[4]
KmGPP 6.0 µM Km value for E.coli E57, from Williams 1998

An excerpt from Günnewich 2006

[4]
KmGPP 12.6 µM Km value for E.coli R58, from Williams 1998

An excerpt from Günnewich 2006

[4]
KmGPP 29.9 µM Km value for E.coli R59, from Williams 1998

An excerpt from Günnewich 2006

[4]
KmGPP 16 µM An excerpt from Williams 1998, results section [4]
KmGPP 6.7 µM Data from Ogura 1997, from wildtype E.coli, as described in Williams 1998. ( I will get the book!)

An excerpt from Williams 1998, results section

[5]

Substrate and Enzyme Concentrations or Amount

Parameter Value Units Description References
[GPP] 40 µM Limonene synthase expressed at 37°C, 40µM GPP, Citrus sinensis, product formation 1.17x105 µM/sec

An excerpt from Olsen 2011 Thesis

[1]
[GPP] 10 µM Limonene synthase expressed at 37°C, 40µM GPP, Citrus sinensis, product formation 1.17x105 µM/sec, [E]: 0.3 µM 
An excerpt from Olsen 2011 Thesis
[1]
[LimSynth] 0.3 µM Limonene synthase expressed at 37°C, 10µM GPP, Citrus sinensis, product formation 1.17x105 µM/sec, [E]: 0.3 µM

An excerpt from Olsen 2011 Thesis

[1]
[LimSynth] 0.07 µM Limonene synthase expressed at 20°C, 200µM GPP, Citrus sinensis

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
[LimSynth] 0.26 µM Limonene synthase expressed at 37°C, 200µM GPP, Citrus sinensis, pH 7.5

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
Enzyme amount 1-10 µg An excerpt from Alonso 1992, Materials and Methods section [3]
Enzyme amount 10-20 µg An excerpt from Adam 1996, Materials and Methods section [8]
[GPP] 10 µM An excerpt from Alonso 1992, Materials and Methods section [3]
[GPP] 2 µM An excerpt from Lucker 2002, Materials and Methods section [7]
[GPP] 0.1 µM 0.1 -100 µM

An excerpt from Lucker 2002, Materials and Methods section

[7]
[GPP] 100 µM 0.1 -100 µM

An excerpt from Lucker 2002, Materials and Methods section

[7]
[LimSynth] 1 - 50 µg Amount of native or recombinant protein (E.coli) used for Limonene synthase enzyme assay

An excerpt from Williams 1998, Materials Method section

[4]
[GPP] 1 - 40 µM Substrate concentration range for enzyme assay

An excerpt from Williams 1998, Materials Method section

[4]
[GPP] 10 µM An excerpt from Günnewich 2006 [6]
Enzyme Amount (Possibly) 1.25 µg Possible it is referring to enzyme amount used for CsTPS1 (limonene producing terpene synthase). 
An excerpt from Günnewich 2006
[6]
[GPP] 25 µM GPP concentration used, for preprotein (?) expressed in E.coli.

An excerpt from Williams 1998, Materials and Methods section

[4]
Enzyme amount 150 µg Purified (-)-4S-limonene synthase amount.

An excerpt from Williams 1998, Materials and Methods section

[4]

Maximum velocity (Vmax), Enzyme activity & Specific enzyme activity

Parameter Value Units Description References
Vmaxforward 0.53 µM/min Limonene synthase expressed at 20°C, 200µM GPP, Citrus sinensis, pH7.5

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
Vmaxforward 0.64 µM/min Limonene synthase expressed at 37°C, 200µM GPP

An excerpt from Entova 2013 Thesis, Chapter 2

[2]
Specific enzyme activity 19 µmol/h/mg An excerpt from Alonso 1992 [3]


Other information

Parameter Value Units Description References
Limonene Synthase

Molecular Weight

56 kDa kDa Limonene synthase purification 
An excerpt from Alonso 1992, Abstract
[3]

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References

  1. ↑ 1.0 1.1 1.2 1.3 Olsen, S. N. 2011. "Isolation, Purification, and Characterization of (+)-4R-limonene synthase: The first step in exploring enzyme stereospecificity in terpenoid biosynthesis", Masters Thesis, Brandeis Univeristy
  2. ↑ 2.0 2.1 2.2 2.3 2.4 2.5 2.6 Entova, S. 2013. "Kinetic characterization, crystallization, and photosynthetic expression of (+)-4R-limonene synthase from C. sinensis", Masters Thesis, Brandeis Univeristy
  3. ↑ 3.0 3.1 3.2 3.3 3.4 Alonso et. al. 1992. "Purification of 4S-Limonene Synthase, a Monoterpene Cyclase from the Glandular Trichomes of Peppermint (Mentha x piperita) and Spearmint (Mentha spicata)", The journal of Biological Chemistry, 267(11):7582-7587
  4. ↑ 4.0 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 Williams, D.C. et. al. 1998. [www.ncbi.nlm.nih.gov/pubmed/9724535 "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 ",Biochemistry, 37:12213-12220] Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content Cite error: Invalid <ref> tag; name "Williams1998" defined multiple times with different content
  5. ↑ 5.0 5.1 Ogura, K. & Koyama, T. 1997. "Dynamic Aspects of Natural Products Chemistry", pp 1-23 Cite error: Invalid <ref> tag; name "Ogura1997" defined multiple times with different content
  6. ↑ 6.0 6.1 6.2 6.3 Cite error: Invalid <ref> tag; name "Gunnewich2006" defined multiple times with different content Cite error: Invalid <ref> tag; name "Gunnewich2006" defined multiple times with different content
  7. ↑ 7.0 7.1 7.2 7.3 Cite error: Invalid <ref> tag; name "Lucker2002" defined multiple times with different content Cite error: Invalid <ref> tag; name "Lucker2002" defined multiple times with different content Cite error: Invalid <ref> tag; name "Lucker2002" defined multiple times with different content
  8. ↑ 8.0 8.1 Adam, K. et. al. 1996. "Partial purification and characterization of a monoterpene cyclase, limonene synthase, from the liverwort Ricciocarpos natans. 332(2):pp 352-356.
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