Difference between revisions of "Limonene Synthase: Detailed information on the kinetic values obtained from the literature"

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==Kinetic Parameters==
 
==Kinetic Parameters==
 +
 +
=== Turnover numbers (Kcat) ===
 
{| class="wikitable sortable"
 
{| class="wikitable sortable"
 
|-
 
|-
Line 18: Line 20:
 
  [[file: 32_LimSynth_kcat_Olsen11.png |400px | An excerpt from Olsen 2011 Thesis ]]
 
  [[file: 32_LimSynth_kcat_Olsen11.png |400px | An excerpt from Olsen 2011 Thesis ]]
 
| <ref name="Olsen2011">Olsen, S. N. 2011. [http://bir.brandeis.edu/handle/10192/24424?show=full "Isolation, Purification, and Characterization of (+)-4R-limonene synthase: The first step in exploring enzyme stereospecificity in terpenoid biosynthesis"], Masters Thesis, Brandeis Univeristy</ref>
 
| <ref name="Olsen2011">Olsen, S. N. 2011. [http://bir.brandeis.edu/handle/10192/24424?show=full "Isolation, Purification, and Characterization of (+)-4R-limonene synthase: The first step in exploring enzyme stereospecificity in terpenoid biosynthesis"], Masters Thesis, Brandeis Univeristy</ref>
 +
|-
 +
| kcat<sub>forward</sub>
 +
| 0.13
 +
| s<sup>-1</sup>
 +
| Limonene synthase expressed at 20°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
 +
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 +
| <ref name="Entova2013"></ref>
 +
|-
 +
| kcat<sub>forward</sub>
 +
| 0.04
 +
| s<sup>-1</sup>
 +
| Limonene synthase expressed at 37°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
 +
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 +
| <ref name="Entova2013"></ref>
 +
|-
 +
| Kcat
 +
| 0.3
 +
| s<sup>-1</sup>
 +
| [[file: 03 LimSynth Kcat Alonso92.png | 450px | An excerpt from Alonso 1992]]
 +
| <ref name="Alonso1992"></ref>
 +
|-
 +
| Kcat
 +
| 0.020
 +
| s<sup>-1</sup>
 +
| [[file: 15_LimSynth_Km_Kcat_Williams98.png| 450px | An excerpt from Williams 1998, Result section]]
 +
| <ref name="Williams1998"> 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]</ref>
 +
|-
 +
| Kcat
 +
| 0.024
 +
| s<sup>-1</sup>
 +
| Below is an excerpt from Williams 1998, but data source is from Ogura 1997, describing data from wildtype ''E.coli''
 +
[[file: 15_LimSynth_Km_Kcat_Williams98.png| 450px | An excerpt from Williams 1998, Result section]]
 +
| <ref name="Ogura1997"> Ogura, K. & Koyama, T. 1997. "Dynamic Aspects of Natural Products Chemistry", pp 1-23 </ref>
 +
|-
 +
| Kcat
 +
| 0.082
 +
| s<sup>-1</sup>
 +
| Kcat value for CsTPS1, ''Cannabis sativa'', pH6.5, 40°C, enzyme assay
 +
[[file: 12_LimSynth_Km_Kcat_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Gunnewich2006"> </ref>
 +
|-
 +
|}
 +
 +
=== Michaelis-Menten constant (Km) ===
 +
{| class="wikitable sortable"
 +
|-
 +
! Parameter
 +
! Value
 +
! Units
 +
! Description
 +
! References
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 130
 +
| µM
 +
| [[file: 26_LimSynth_Km_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 3 ]]
 +
| <ref name="Entova2013"></ref>
 +
|-
 +
| Km
 +
| 0.7
 +
| µM
 +
| [[file: 28_LimSynth_Km_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
 +
| <ref name="Lucker2002">  </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 1.25
 +
| µM
 +
| [[file: 18_LimSynth_GPP_conc_Adan96.png | 400px | An excerpt from Adam 1996, Materials and Methods section]]
 +
| <ref name="Adam1996"> 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.</ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 6.8
 +
| µM
 +
| Km value for CsTPS1, pH6.5, 40°C, ''Cannabis sativa'' enzyme assay
 +
[[file: 12_LimSynth_Km_Kcat_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Gunnewich2006"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 8.6
 +
| µM
 +
| Km value for E.coli ''Q53'', from Williams 1998
 +
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Williams1998"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 6.0
 +
| µM
 +
| Km value for E.coli ''E57'', from Williams 1998
 +
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Williams1998"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 12.6
 +
| µM
 +
| Km value for E.coli ''R58'', from Williams 1998
 +
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Williams1998"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 29.9
 +
| µM
 +
| Km value for E.coli ''R59'', from Williams 1998
 +
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 +
| <ref name="Williams1998"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 16
 +
| µM
 +
| [[file: 15_LimSynth_Km_Kcat_Williams98.png | 300px | An excerpt from Williams 1998, results section]]
 +
| <ref name="Williams1998"> </ref>
 +
|-
 +
| Km<sub>GPP</sub>
 +
| 6.7
 +
| µM
 +
| Data from Ogura 1997, from wildtype ''E.coli'', as described in Williams 1998. ( I will get the book!)
 +
[[file: 15_LimSynth_Km_Kcat_Williams98.png | 300px | An excerpt from Williams 1998, results section]]
 +
| <ref name="Ogura1997"> </ref>
 +
|-
 +
|}
 +
 +
=== Substrate and Enzyme Concentrations or Amount ===
 +
{| class="wikitable sortable"
 +
|-
 +
! Parameter
 +
! Value
 +
! Units
 +
! Description
 +
! References
 
|-
 
|-
 
| [GPP]
 
| [GPP]
Line 33: Line 163:
 
| <ref name="Olsen2011"></ref>
 
| <ref name="Olsen2011"></ref>
 
|-
 
|-
−
| [Limonene synthase]
+
| [LimSynth]
 
| 0.3  
 
| 0.3  
 
| µM
 
| µM
Line 46: Line 176:
 
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
| <ref name="Entova2013">Entova, S. 2013. [http://bir.brandeis.edu/handle/10192/25158 "Kinetic characterization, crystallization, and photosynthetic expression of (+)-4R-limonene synthase from ''C. sinensis''"], Masters Thesis, Brandeis Univeristy</ref>
 
| <ref name="Entova2013">Entova, S. 2013. [http://bir.brandeis.edu/handle/10192/25158 "Kinetic characterization, crystallization, and photosynthetic expression of (+)-4R-limonene synthase from ''C. sinensis''"], Masters Thesis, Brandeis Univeristy</ref>
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 130
 
−
| µM
 
−
| [[file: 26_LimSynth_Km_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 3 ]]
 
−
| <ref name="Entova2013"></ref>
 
 
|-
 
|-
 
| [LimSynth]
 
| [LimSynth]
Line 57: Line 181:
 
| µM
 
| µM
 
|  Limonene synthase expressed at 37°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
 
|  Limonene synthase expressed at 37°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
−
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
−
| <ref name="Entova2013"></ref>
 
−
|-
 
−
| Vmax<sub>forward</sub>
 
−
| 0.53
 
−
| µM/min
 
−
|  Limonene synthase expressed at 20°C, 200µM GPP, ''Citrus sinensis'', pH7.5
 
−
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
−
| <ref name="Entova2013"></ref>
 
−
|-
 
−
| Vmax<sub>forward</sub>
 
−
|0.64
 
−
| µM/min
 
−
| Limonene synthase expressed at 37°C, 200µM GPP
 
−
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
−
| <ref name="Entova2013"></ref>
 
−
|-
 
−
| kcat<sub>forward</sub>
 
−
| 0.13
 
−
| s<sup>-1</sup>
 
−
| Limonene synthase expressed at 20°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
 
−
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
−
| <ref name="Entova2013"></ref>
 
−
|-
 
−
| kcat<sub>forward</sub>
 
−
| 0.04
 
−
| s<sup>-1</sup>
 
−
| Limonene synthase expressed at 37°C, 200µM GPP, ''Citrus sinensis'', pH 7.5
 
 
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 
| <ref name="Entova2013"></ref>
 
| <ref name="Entova2013"></ref>
Line 117: Line 213:
 
| 0.1 -100 µM
 
| 0.1 -100 µM
 
[[file: 27_LimSynth_GPP_conc_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
 
[[file: 27_LimSynth_GPP_conc_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
−
| <ref name="Lucker2002">  </ref>
 
−
|-
 
−
| Km
 
−
| 0.7
 
−
| µM
 
−
| [[file: 28_LimSynth_Km_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
 
 
| <ref name="Lucker2002">  </ref>
 
| <ref name="Lucker2002">  </ref>
 
|-
 
|-
Line 131: Line 221:
 
[[file: 27_LimSynth_GPP_conc_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
 
[[file: 27_LimSynth_GPP_conc_Lucker02.png | 400px | An excerpt from Lucker 2002, Materials and Methods section]]
 
| <ref name="Lucker2002">  </ref>
 
| <ref name="Lucker2002">  </ref>
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 1.25
 
−
| µM
 
−
| [[file: 18_LimSynth_GPP_conc_Adan96.png | 400px | An excerpt from Adam 1996, Materials and Methods section]]
 
−
| <ref name="Adam1996"> 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.</ref>
 
−
|-
 
−
| Specific enzyme activity
 
−
| 19
 
−
| µmol/h/mg
 
−
| [[file: 03 LimSynth Kcat Alonso92.png | 450px | An excerpt from Alonso 1992]]
 
−
| <ref name="Alonso1992"></ref>
 
−
|-
 
−
| Kcat
 
−
| 0.3
 
−
| s<sup>-1</sup>
 
−
| [[file: 03 LimSynth Kcat Alonso92.png | 450px | An excerpt from Alonso 1992]]
 
−
| <ref name="Alonso1992"></ref>
 
−
|-
 
−
| Kcat
 
−
| 0.020
 
−
| s<sup>-1</sup>
 
−
| [[file: 15_LimSynth_Km_Kcat_Williams98.png| 450px | An excerpt from Williams 1998, Result section]]
 
−
| <ref name="Williams1998"> 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]</ref>
 
 
|-
 
|-
 
| [LimSynth]
 
| [LimSynth]
Line 169: Line 235:
 
[[file: 30_LimSynth_GPP_conc_Williams98.png| 450px | An excerpt from Williams 1998, Materials Method section]]
 
[[file: 30_LimSynth_GPP_conc_Williams98.png| 450px | An excerpt from Williams 1998, Materials Method section]]
 
| <ref name="Williams1998"> </ref>
 
| <ref name="Williams1998"> </ref>
−
|-
 
−
| Kcat
 
−
| 0.024
 
−
| s<sup>-1</sup>
 
−
| Below is an excerpt from Williams 1998, but data source is from Ogura 1997, describing data from wildtype ''E.coli''
 
−
[[file: 15_LimSynth_Km_Kcat_Williams98.png| 450px | An excerpt from Williams 1998, Result section]]
 
−
| <ref name="Ogura1997"> Ogura, K. & Koyama, T. 1997. "Dynamic Aspects of Natural Products Chemistry", pp 1-23 </ref>
 
 
|-
 
|-
 
| [GPP]
 
| [GPP]
Line 188: Line 247:
 
| Possible it is referring to enzyme amount used for CsTPS1 (limonene producing terpene synthase).
 
| Possible it is referring to enzyme amount used for CsTPS1 (limonene producing terpene synthase).
 
  [[file: 11_LimSynth_possible_enzyme_amount_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
 
  [[file: 11_LimSynth_possible_enzyme_amount_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
−
| <ref name="Gunnewich2006"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 6.8
 
−
| µM
 
−
| Km value for CsTPS1, pH6.5, 40°C, ''Cannabis sativa'' enzyme assay
 
−
[[file: 12_LimSynth_Km_Kcat_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
 
−
| <ref name="Gunnewich2006"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 8.6
 
−
| µM
 
−
| Km value for E.coli ''Q53'', from Williams 1998
 
−
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 
−
| <ref name="Williams1998"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 6.0
 
−
| µM
 
−
| Km value for E.coli ''E57'', from Williams 1998
 
−
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 
−
| <ref name="Williams1998"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 12.6
 
−
| µM
 
−
| Km value for E.coli ''R58'', from Williams 1998
 
−
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 
−
| <ref name="Williams1998"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 29.9
 
−
| µM
 
−
| Km value for E.coli ''R59'', from Williams 1998
 
−
[[file: 16_LimSynth_EColi_Km_Kcat_Williams98.png | 300px | An excerpt from Günnewich 2006]]
 
−
| <ref name="Williams1998"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 16
 
−
| µM
 
−
| [[file: 15_LimSynth_Km_Kcat_Williams98.png | 300px | An excerpt from Williams 1998, results section]]
 
−
| <ref name="Williams1998"> </ref>
 
−
|-
 
−
| Km<sub>GPP</sub>
 
−
| 6.7
 
−
| µM
 
−
| Data from Ogura 1997, from wildtype ''E.coli'', as described in Williams 1998. ( I will get the book!)
 
−
[[file: 15_LimSynth_Km_Kcat_Williams98.png | 300px | An excerpt from Williams 1998, results section]]
 
−
| <ref name="Ogura1997"> </ref>
 
−
|-
 
−
| Kcat
 
−
| 0.082
 
−
| s<sup>-1</sup>
 
−
| Kcat value for CsTPS1, ''Cannabis sativa'', pH6.5, 40°C, enzyme assay
 
−
[[file: 12_LimSynth_Km_Kcat_Gunnewich06.png | 300px | An excerpt from Günnewich 2006]]
 
 
| <ref name="Gunnewich2006"> </ref>
 
| <ref name="Gunnewich2006"> </ref>
 
|-
 
|-
Line 261: Line 265:
 
|}
 
|}
  
−
[http://www.systemsbiology.ls.manchester.ac.uk/wiki/index.php/Kinetic_Model_of_Monoterpenoid_Biosynthesis_Wiki Back to the main model page].
+
=== Maximum velocity (Vmax), Enzyme activity & Specific enzyme activity ===
 +
{| class="wikitable sortable"
 +
|-
 +
! Parameter
 +
! Value
 +
! Units
 +
! Description
 +
! References
 +
|-
 +
| Vmax<sub>forward</sub>
 +
| 0.53
 +
| µM/min
 +
|  Limonene synthase expressed at 20°C, 200µM GPP, ''Citrus sinensis'', pH7.5
 +
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 +
| <ref name="Entova2013"></ref>
 +
|-
 +
| Vmax<sub>forward</sub>
 +
|0.64
 +
| µM/min
 +
| Limonene synthase expressed at 37°C, 200µM GPP
 +
[[file: 25_LimSynth_kinetics_Entova13.png |400px | An excerpt from Entova 2013 Thesis, Chapter 2 ]]
 +
| <ref name="Entova2013"></ref>
 +
|-
 +
| Specific enzyme activity
 +
| 19
 +
| µmol/h/mg
 +
| [[file: 03 LimSynth Kcat Alonso92.png | 450px | An excerpt from Alonso 1992]]
 +
| <ref name="Alonso1992"></ref>
 +
|-
 +
|}
 +
 
 +
 
  
 
==Other information==
 
==Other information==
Line 282: Line 317:
 
|-
 
|-
 
|}
 
|}
 +
 +
[http://www.systemsbiology.ls.manchester.ac.uk/wiki/index.php/Kinetic_Model_of_Monoterpenoid_Biosynthesis_Wiki Back to the main model page].
  
 
==References ==
 
==References ==
 
<references />
 
<references />

Revision as of 15:41, 14 April 2016

Back to terpene model MAIN page.

Back to LimSynth model page.

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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