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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
|
[1]
|
kcatforward
|
0.13
|
s-1
|
Limonene synthase expressed at 20°C, 200µM GPP, Citrus sinensis, pH 7.5
|
[2]
|
kcatforward
|
0.04
|
s-1
|
Limonene synthase expressed at 37°C, 200µM GPP, Citrus sinensis, pH 7.5
|
[2]
|
Kcat
|
0.3
|
s-1
|
Limonene synthase assay- Mentha species, pH7.0, 30°C, typical reaction mixture contained 1-10 μg protein and 15mM MgCl2, Initial [GPP} is 10μM
|
[3]
|
Kcat
|
0.020
|
s-1
|
|
[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
|
[5]
|
Kcat
|
0.082
|
s-1
|
Kcat value for CsTPS1, Cannabis sativa, pH6.5, 40°C, enzyme assay
|
[6]
|
Michaelis-Menten constant (Km)
Parameter
|
Value
|
Units
|
Description
|
References
|
KmGPP
|
6.25 ± 0.41
|
µM
|
CsTPS1 (limonene producing), Km of 6.25 ± 0.41 mM, pH 6.5, 40°C, [GPP] 0.5 - 25.0 mM used for kinetic parameter determination, kcat 0.09 s-1
|
[7]
|
KmGPP
|
130
|
µM
|
|
[2]
|
Km
|
0.7
|
µM
|
|
[8]
|
KmGPP
|
1.25
|
µM
|
|
[9]
|
KmGPP
|
6.8
|
µM
|
Km value for CsTPS1, pH6.5, 40°C, Cannabis sativa enzyme assay
|
[6]
|
KmGPP
|
8.6
|
µM
|
Km value for E.coli Q53, from Williams 1998
|
[4]
|
KmGPP
|
6.0
|
µM
|
Km value for E.coli E57, from Williams 1998
|
[4]
|
KmGPP
|
12.6
|
µM
|
Km value for E.coli R58, from Williams 1998
|
[4]
|
KmGPP
|
29.9
|
µM
|
Km value for E.coli R59, from Williams 1998
|
[4]
|
KmGPP
|
16
|
µM
|
|
[4]
|
KmGPP
|
6.7
|
µM
|
Data from Ogura 1997, from wildtype E.coli, as described in Williams 1998. ( I will get the book!)
|
[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
|
[1]
|
[GPP]
|
10
|
µM
|
Limonene synthase expressed at 37°C, 40µM GPP, Citrus sinensis, product formation 1.17x105 µM/sec, [E]: 0.3 µM
|
[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
|
[1]
|
[LimSynth]
|
0.07
|
µM
|
Limonene synthase expressed at 20°C, 200µM GPP, Citrus sinensis
|
[2]
|
[LimSynth]
|
0.26
|
µM
|
Limonene synthase expressed at 37°C, 200µM GPP, Citrus sinensis, pH 7.5
|
[2]
|
Enzyme amount
|
1-10
|
µg
|
|
[3]
|
Enzyme amount
|
10-20
|
µg
|
|
[9]
|
[GPP]
|
10
|
µM
|
|
[3]
|
[GPP]
|
2
|
µM
|
|
[8]
|
[GPP]
|
0.1
|
µM
|
0.1 -100 µM
|
[8]
|
[GPP]
|
100
|
µM
|
0.1 -100 µM
|
[8]
|
[LimSynth]
|
1 - 50
|
µg
|
Amount of native or recombinant protein (E.coli) used for Limonene synthase enzyme assay
|
[4]
|
[GPP]
|
1 - 40
|
µM
|
Substrate concentration range for enzyme assay
|
[4]
|
[GPP]
|
10
|
µM
|
|
[6]
|
Enzyme Amount (Possibly)
|
1.25
|
µg
|
Possible it is referring to enzyme amount used for CsTPS1 (limonene producing terpene synthase).
|
[6]
|
[GPP]
|
25
|
µM
|
GPP concentration used, for preprotein (?) expressed in E.coli.
|
[4]
|
Enzyme amount
|
150
|
µg
|
Purified (-)-4S-limonene synthase amount.
|
[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
|
[2]
|
Vmaxforward
|
0.64
|
µM/min
|
Limonene synthase expressed at 37°C, 200µM GPP
|
[2]
|
Specific enzyme activity
|
19
|
µmol/h/mg
|
|
[3]
|
Other information
Parameter
|
Value
|
Units
|
Description
|
References
|
Limonene Synthase
Molecular Weight
|
56 kDa
|
kDa
|
Limonene synthase purification
|
[3]
|
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References
- ↑ 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
- ↑ 2.0 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
- ↑ 3.0 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
- ↑ 4.0 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
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- ↑ 5.0 5.1 Ogura, K. & Koyama, T. 1997. "Dynamic Aspects of Natural Products Chemistry", pp 1-23 Cite error: Invalid
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- ↑ 6.0 6.1 6.2 6.3 Cite error: Invalid
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- ↑ Gunnewich, N. 2008 "Expression and characterization of terpene synthases from Cannabis sativa L. and Salvia sclarea L.
- ↑ 8.0 8.1 8.2 8.3 Cite error: Invalid
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- ↑ 9.0 9.1 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.