Difference between revisions of "Part:BBa K3038003"

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Thioesterase III (FadM) is a long-chain acyl-CoA thioesterase that is involved in the β-oxidation of oleic acid. The enzyme is able to hydrolyze a number of related substrates. The best substrate is 3,5-tetradecadienoyl-CoA, which is a minor side product of oleate β-oxidation that is resistant to further degradation. The hydrolysis product, 3,5-tetradecadienoate, is released into the growth medium [Ren04a, Nie08]. Thioesterase III is expressed upon growth on oleic acid as the sole source of carbon [Ren04a, Nie08]. FadM is a member of the fad regulon; expression is induced by a number of fatty acids, with C18:1 as the best inducer [Feng09b]. Reports disagree on whether [Nie08a] or not [Feng09b] conjugated linoleic acid (CLA) induces an even higher level of expression of fadM.
 
Thioesterase III (FadM) is a long-chain acyl-CoA thioesterase that is involved in the β-oxidation of oleic acid. The enzyme is able to hydrolyze a number of related substrates. The best substrate is 3,5-tetradecadienoyl-CoA, which is a minor side product of oleate β-oxidation that is resistant to further degradation. The hydrolysis product, 3,5-tetradecadienoate, is released into the growth medium [Ren04a, Nie08]. Thioesterase III is expressed upon growth on oleic acid as the sole source of carbon [Ren04a, Nie08]. FadM is a member of the fad regulon; expression is induced by a number of fatty acids, with C18:1 as the best inducer [Feng09b]. Reports disagree on whether [Nie08a] or not [Feng09b] conjugated linoleic acid (CLA) induces an even higher level of expression of fadM.
  
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https://biocyc.org/gene?orgid=ECOLI&id=G6244
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https://biocyc.org/gene?orgid=ECOLI&id=G6244<br/>
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FadM_Cter-FLAG
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For the competition, FadM is tagged in Cterm by a FLAG tag. This tag allows the purification of the protein in order to test the activity.
  
 
===GenBank===
 
===GenBank===
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===Protein Sequence ===     
 
===Protein Sequence ===     
  
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        10        20        30        40        50
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Without the Flag-tag :<br/>
 
MQTQIKVRGY HLDVYQHVNN ARYLEFLEEA RWDGLENSDS FQWMTAHNIA  
 
MQTQIKVRGY HLDVYQHVNN ARYLEFLEEA RWDGLENSDS FQWMTAHNIA  
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        60        70        80        90        100
 
 
FVVVNININY RRPAVLSDLL TITSQLQQLN GKSGILSQVI TLEPEGQVVA  
 
FVVVNININY RRPAVLSDLL TITSQLQQLN GKSGILSQVI TLEPEGQVVA  
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      110        120        130
 
 
DALITFVCID LKTQKALALE GELREKLEQM VK
 
DALITFVCID LKTQKALALE GELREKLEQM VK
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===Ecoli Long chain acyl CoA thioesterase ===
 
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===Thioesterase III===
 
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Thioesterase III (FadM) is a long-chain acyl-CoA thioesterase that is involved in the β-oxidation of oleic acid. The enzyme is able to hydrolyze a number of related substrates. The best substrate is 3,5-tetradecadienoyl-CoA, which is a minor side product of oleate β-oxidation that is resistant to further degradation. The hydrolysis product, 3,5-tetradecadienoate, is released into the growth medium [Ren04a, Nie08].
 
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Thioesterase III is expressed upon growth on oleic acid as the sole source of carbon [Ren04a, Nie08]. fadM is a member of the fad regulon; expression is induced by a number of fatty acids, with C18:1 as the best inducer [Feng09b]. Reports disagree on whether [Nie08a] or not [Feng09b] conjugated linoleic acid (CLA) induces an even higher level of expression of fadM.
 
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https://biocyc.org/gene?orgid=ECOLI&id=G6244
 
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===Protein Sequence===
 
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(not tagged)
 
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MQTQIKVRGY HLDVYQHVNN ARYLEFLEEA RWDGLENSDS FQWMTAHNIA
 
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FVVVNININY RRPAVLSDLL TITSQLQQLN GKSGILSQVI TLEPEGQVVA
 
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DALITFVCID LKTQKALALE GELREKLEQM VK
 
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Molecular size : 15.088 kD (from nucleotide sequence)
 
Molecular size : 15.088 kD (from nucleotide sequence)
  
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===Reference===
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==Reference==
  
 
Engineering of Bacterial Methyl Ketone Synthesis for Biofuels. Ee-Been Goh,a,c Edward E. K. Baidoo,a,c Jay D. Keasling,a,c,d and Harry R. Beller. Appl Environ Microbiol. 2012 Jan; 78(1): 70–80. doi: 10.1128/AEM.06785-11. PMCID: PMC3255637. PMID: 22038610
 
Engineering of Bacterial Methyl Ketone Synthesis for Biofuels. Ee-Been Goh,a,c Edward E. K. Baidoo,a,c Jay D. Keasling,a,c,d and Harry R. Beller. Appl Environ Microbiol. 2012 Jan; 78(1): 70–80. doi: 10.1128/AEM.06785-11. PMCID: PMC3255637. PMID: 22038610
  
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===Usage and Biology===
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==Usage and Biology==
  
 
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Revision as of 08:22, 19 October 2019

Description

FadM is for E. coli long-chain acyl CoA thioesterase that is Thioesterase III.
Thioesterase III (FadM) is a long-chain acyl-CoA thioesterase that is involved in the β-oxidation of oleic acid. The enzyme is able to hydrolyze a number of related substrates. The best substrate is 3,5-tetradecadienoyl-CoA, which is a minor side product of oleate β-oxidation that is resistant to further degradation. The hydrolysis product, 3,5-tetradecadienoate, is released into the growth medium [Ren04a, Nie08]. Thioesterase III is expressed upon growth on oleic acid as the sole source of carbon [Ren04a, Nie08]. FadM is a member of the fad regulon; expression is induced by a number of fatty acids, with C18:1 as the best inducer [Feng09b]. Reports disagree on whether [Nie08a] or not [Feng09b] conjugated linoleic acid (CLA) induces an even higher level of expression of fadM.

https://biocyc.org/gene?orgid=ECOLI&id=G6244
For the competition, FadM is tagged in Cterm by a FLAG tag. This tag allows the purification of the protein in order to test the activity.

GenBank

FadM : GenBank: P77712
https://www.uniprot.org/uniprot/P77712

Protein Sequence

Without the Flag-tag :
MQTQIKVRGY HLDVYQHVNN ARYLEFLEEA RWDGLENSDS FQWMTAHNIA FVVVNININY RRPAVLSDLL TITSQLQQLN GKSGILSQVI TLEPEGQVVA DALITFVCID LKTQKALALE GELREKLEQM VK

Molecular size : 15.088 kD (from nucleotide sequence)

Reference

Engineering of Bacterial Methyl Ketone Synthesis for Biofuels. Ee-Been Goh,a,c Edward E. K. Baidoo,a,c Jay D. Keasling,a,c,d and Harry R. Beller. Appl Environ Microbiol. 2012 Jan; 78(1): 70–80. doi: 10.1128/AEM.06785-11. PMCID: PMC3255637. PMID: 22038610

Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]