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Literature summary for 2.3.1.304 extracted from

  • Jia, K.; Cao, R.; Hua, D.H.; Li, P.
    Study of class I and class III polyhydroxyalkanoate (PHA) synthases with substrates containing a modified side chain (2016), Biomacromolecules, 17, 1477-1485 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.3.1.304

Activating Compound

Activating Compound Comment Organism Structure
bovine serum albumin addition of BSA increases the specific activity of enzyme PhaCCs with HBCoA about 2fold Chromobacterium sp. USM2
bovine serum albumin leads to an activity increase of severalfold when BSA is added to a concentration of 0.5 mg/mL (0.0075 mM) in the assay mixture Allochromatium vinosum
additional information molecular activation mechanism, overview Allochromatium vinosum
additional information molecular activation mechanism, overview Chromobacterium sp. USM2
additional information no significant activation of enzyme PhaCCc by bovine serum albumin, molecular activation mechanism, overview Caulobacter vibrioides

Cloned(Commentary)

Cloned (Comment) Organism
enzyme PhaCCc, recombinant expression Caulobacter vibrioides
enzyme PhaECAv, recombinant expression Allochromatium vinosum
gene phaC, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Chromobacterium sp. USM2

Protein Variants

Protein Variants Comment Organism
A479S site-directed mutagenesis, while mutant A479S-PhaCCs displays the same order as the wild-type enzyme with regard to chain length, it exhibits higher activity for both HBCoA and HVCoA and lower activity for HHxCoA compared to wild-type Chromobacterium sp. USM2

Inhibitors

Inhibitors Comment Organism Structure
additional information no inhibition by bovine serum albumin Allochromatium vinosum
additional information no inhibition by bovine serum albumin Caulobacter vibrioides
additional information no inhibition by bovine serum albumin Chromobacterium sp. USM2

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Allochromatium vinosum
additional information
-
 additional information Michaelis-Menten kinetics Chromobacterium sp. USM2
additional information
-
 additional information Michaelis-Menten kinetics. Caulobacter crescentus class I PHA synthase uniquely lacks of a lag phase in its polymerization kinetics Caulobacter vibrioides
0.05
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
0.05
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
0.07
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
0.11
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
0.11
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
0.29
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
1.54
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
2.07
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
3.22
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
9.46
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(R)-3-hydroxybutanoyl-CoA + [(R)-3-hydroxybutanoate]n Allochromatium vinosum
-
 [(R)-3-hydroxybutanoate](n+1) + CoA
-
 ?

Organism

Organism UniProt Comment Textmining
Allochromatium vinosum
-
-
-
Caulobacter vibrioides A0A290N3S7
-
-
Chromobacterium sp. USM2 E1APK1
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme PhaCCc Caulobacter vibrioides
recombinant enzyme PhaECAv Allochromatium vinosum
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration Chromobacterium sp. USM2

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-3-hydroxybutanoyl-CoA + [(R)-3-hydroxybutanoate]n
-
 Allochromatium vinosum [(R)-3-hydroxybutanoate](n+1) + CoA
-
 ?
(R)-3-hydroxybutanoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HBCoA Allochromatium vinosum [(R)-3-hydroxybutanoate](n+1) + CoA
-
 ?
(R)-3-hydroxybutanoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HBCoA Chromobacterium sp. USM2 [(R)-3-hydroxybutanoate](n+1) + CoA
-
 ?
(R)-3-hydroxybutanoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HBCoA Caulobacter vibrioides [(R)-3-hydroxybutanoate](n+1) + CoA
-
 ?
(R)-3-hydroxycapryl-CoA + [(R)-3-hydroxybutanoate]n substrate HCCoA Allochromatium vinosum ? + CoA
-
 ?
(R)-3-hydroxycapryl-CoA + [(R)-3-hydroxybutanoate]n substrate HCCoA Chromobacterium sp. USM2 ? + CoA
-
 ?
(R)-3-hydroxycapryl-CoA + [(R)-3-hydroxybutanoate]n substrate HCCoA Caulobacter vibrioides ? + CoA
-
 ?
(R)-3-hydroxyvaleryl-CoA + [(R)-3-hydroxybutanoate]n substrate HVCoA Allochromatium vinosum ? + CoA
-
 ?
(R)-3-hydroxyvaleryl-CoA + [(R)-3-hydroxybutanoate]n substrate HVCoA Chromobacterium sp. USM2 ? + CoA
-
 ?
(R)-3-hydroxyvaleryl-CoA + [(R)-3-hydroxybutanoate]n substrate HVCoA Caulobacter vibrioides ? + CoA
-
 ?
3-(R)-hydroxhex-5-enoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxeCoA Allochromatium vinosum ? + CoA
-
 ?
3-(R)-hydroxhex-5-enoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxeCoA Chromobacterium sp. USM2 ? + CoA
-
 ?
3-(R)-hydroxhex-5-enoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxeCoA, low activity Caulobacter vibrioides ? + CoA
-
 ?
3-(R)-hydroxyhex5-ynoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxyCoA Allochromatium vinosum ? + CoA
-
 ?
3-(R)-hydroxyhex5-ynoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxyCoA Chromobacterium sp. USM2 ? + CoA
-
 ?
3-(R)-hydroxyhex5-ynoyl-CoA + [(R)-3-hydroxybutanoate]n substrate HHxyCoA, low activity Caulobacter vibrioides ? + CoA
-
 ?
3-(S)-hydroxy-4-azidobutyryl-CoA + [(R)-3-hydroxybutanoate]n substrate HABCoA Allochromatium vinosum ? + CoA
-
 ?
3-(S)-hydroxy-4-azidobutyryl-CoA + [(R)-3-hydroxybutanoate]n substrate HABCoA Chromobacterium sp. USM2 ? + CoA
-
 ?
3-(S)-hydroxy-4-azidobutyryl-CoA + [(R)-3-hydroxybutanoate]n substrate HABCoA, low activity Caulobacter vibrioides ? + CoA
-
 ?
additional information synthesis of a series of 3-(R)-hydroxyacyl CoA (HACoA) analogues as enzyme substrates, substrate specificity compared to class I PHA synthases, overview. The HHxyCoA and HABCoA can be efficiently incorporated into the polymers produced by enzyme PhaECAv. HHxCoA can be metabolically generated from 3-(R)-hydroxy-5-hexynoic acid. The activity of PhaECAv drops significantly with increasing length of the side chain in substrates. For example, the polymerization rates of HVCoA, HHxCoA, and HCCoA catalyzed by PhaECAv are measured at 23%, 0.38%, and 0.005% rate of HBCoA, respectively. No or poor activity with 3-(R)-hydroxy-4-phenylbutyryl-CoA (HPBCoA). Class III PhaECAv can polymerize HABCoA 6.5fold faster than HHxyCoA Allochromatium vinosum ?
-
-
additional information synthesis of a series of 3-(R)-hydroxyacyl CoA (HACoA) analogues as enzyme substrates, substrate specificity compared to class III PHA synthase, overview. PhaCCc displays 2.5fold lower activity with HABCoA than with HHxyCoA Caulobacter vibrioides ?
-
-
additional information synthesis of a series of 3-(R)-hydroxyacyl CoA (HACoA) analogues as enzyme substrates, substrate specificity compared to class III PHA synthase, overview. Priming of PhaCCc with saturated trimeric HBCoA (named sTCoA) demonstrates that approximately one equivalent CoA per PhaC is released during enzyme assay. This is quite different from other reported class I synthases. Wild-type PhaCCs has similar activities toward HHxyCoA and HABCoA Chromobacterium sp. USM2 ?
-
-

Synonyms

Synonyms Comment Organism
class I PHA synthase
-
 Chromobacterium sp. USM2
class I PHA synthase
-
 Caulobacter vibrioides
class I polyhydroxyalkanoate synthase
-
 Chromobacterium sp. USM2
class I polyhydroxyalkanoate synthase
-
 Caulobacter vibrioides
Class I synthase
-
 Chromobacterium sp. USM2
Class I synthase
-
 Caulobacter vibrioides
class III PHA synthase
-
 Allochromatium vinosum
class III polyhydroxyalkanoate synthase
-
 Allochromatium vinosum
class III synthase
-
 Allochromatium vinosum
PHA synthase
-
 Chromobacterium sp. USM2
PHA synthase
-
 Caulobacter vibrioides
PhaC
-
 Chromobacterium sp. USM2
PhaC
-
 Caulobacter vibrioides
PhaCCc
-
 Caulobacter vibrioides
PhaCCs
-
 Chromobacterium sp. USM2
PhaECAv
-
 Allochromatium vinosum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Allochromatium vinosum
30
-
 assay at Chromobacterium sp. USM2
30
-
 assay at Caulobacter vibrioides

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.06
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
0.13
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
0.19
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
2.99
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
12.7
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
14.4
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
20
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
75
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
98.1
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
508
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
 assay at Allochromatium vinosum
7.8
-
 assay at Chromobacterium sp. USM2
7.8
-
 assay at Caulobacter vibrioides

General Information

General Information Comment Organism
physiological function different PHA synthases display distinct preference with regard to the length of the alkyl side chains, they can withstand moderate side chain modifications such as terminal unsaturated bonds and the azide group Allochromatium vinosum
physiological function different PHA synthases display distinct preference with regard to the length of the alkyl side chains, they can withstand moderate side chain modifications such as terminal unsaturated bonds and the azide group Caulobacter vibrioides
physiological function different PHA synthases display distinct preference with regard to the length of the alkyl side chains, they can withstand moderate side chain modifications such as terminal unsaturated bonds and the azide group. Specifically, the specific activity of PhaCCs toward propynyl analogue (HHxyCoA) is only 5fold less than that toward the classical substrate HBCoA Chromobacterium sp. USM2

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.019
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
0.02
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
0.084
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
6.96
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
59.8
-
 3-(R)-hydroxyhex5-ynoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
254
-
 3-(R)-hydroxhex-5-enoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
258.6
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Caulobacter vibrioides
285.7
-
 3-(S)-hydroxy-4-azidobutyryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
891.8
-
 (R)-3-hydroxyvaleryl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum
4618.2
-
 (R)-3-hydroxybutanoyl-CoA recombinant enzyme, pH 7.8, 30°C Allochromatium vinosum