Literature summary for 2.3.1.304 extracted from

Fluechter, S.; Follonier, S.; Schiel-Bengelsdorf, B.; Bengelsdorf, F.R.; Zinn, M.
Anaerobic production of poly(3-hydroxybutyrate) and its precursor 3?hydroxybutyrate from synthesis gas by autotrophic clostridia (2019), Biomacromolecules, 20, 3271-3282 .

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Application

Application	Comment	Organism
synthesis	poly(3-hydroxyalkanoates) (PHAs) such as poly(3-hydroxybutyrate) (PHB) are suitable biobased and biodegradable candidates to replace petroleum-based nondegradable plastics. Enzyme PhaEC can be used in its biocatalytic production, PHB production in clostridia, method development and evaluation, overview. The successful transfer and expression of phaJ and phaEC in autotrophic gas-fermenting clostridia leading to PHB formation now opens the possibility to establish an economically viable route to biodegradable plastics from waste and greenhouse gases, as the gas fermentation technology has meanwhile matured and is already performed at industrial scale. A novel metabolic pathway leading to 3-hydroxybutyrate has also been engineered that will allow similarly better economic production of this platform chemical	Clostridium acetireducens

Cloned(Commentary)

Cloned (Comment)	Organism
gene PhaEC, sequence comparisons, construction of a plasmid with genes encoding thiolase A (thlA), 3-hydroxybutyryl-CoA dehydrogenase (hbd), and crotonase (crt) from Clostridium scatologenes strain ATCC 25775 and (R)-enoyl-CoA hydratase (phaJ) and PHA synthase (phaEC) from Clostridium acetireducens strain DSM 10703 under control of promoter Ppta-ack from Clostridium ljungdahlii strain DSM 13583 into pMTL83151 Clostridium-Escherichia coli shuttle plasmid. The clustered genes thlA, hbd, and crt are initially cloned into pMTL83151, together with Ppta-ack promoter, recombinant expression in Escherichia coli	Clostridium acetireducens

Cloned (Comment)

Organism

gene PhaEC, sequence comparisons, construction of a plasmid with genes encoding thiolase A (thlA), 3-hydroxybutyryl-CoA dehydrogenase (hbd), and crotonase (crt) from Clostridium scatologenes strain ATCC 25775 and (R)-enoyl-CoA hydratase (phaJ) and PHA synthase (phaEC) from Clostridium acetireducens strain DSM 10703 under control of promoter Ppta-ack from Clostridium ljungdahlii strain DSM 13583 into pMTL83151 Clostridium-Escherichia coli shuttle plasmid. The clustered genes thlA, hbd, and crt are initially cloned into pMTL83151, together with Ppta-ack promoter, recombinant expression in Escherichia coli

Clostridium acetireducens

Protein Variants

Protein Variants	Comment	Organism
additional information	anaerobic production of the biopolymer poly(3-hydroxybutyrate) (PHB) and the monomer 3-hydroxybutyrate (3-HB) is achieved using recombinant clostridial acetogens supplied with syn-(thesis) gas as the sole carbon and energy source. 3-HB production is successfully accomplished by a synthetic pathway containing the genes thlA (encoding thiolase A), ctfA/B (encoding CoA-transferase A/B), and bdhA (encoding (R)-3-hydroxybutyrate dehydrogenase). The respective recombinant Clostridium coskatii [p83_tcb] strain produces autotrophically and heterotrophically 3-HB. As a proof of concept, production of PHB is achieved using recombinant Clostridium coskatii and Clostridium ljungdahlii strains expressing a synthetic PHB pathway containing the genes thlA (encoding thiolase A), hbd (encoding 3-hydroxybutyryl-CoA dehydrogenase), crt (encoding crotonase), phaJ (encoding (R)-enoyl-CoA hydratase), and phaEC (encoding PHA synthase). The strain Clostridium coskatii [p83_PHB_Scaceti] synthesizes heterotrophically 3.4% PHB per cell dry weight (CDW) and autotrophically 1.12% PHB per CDW	Clostridium acetireducens

Organism

Organism	UniProt	Comment	Textmining
Clostridium acetireducens	A0A1E8EW93 AND A0A1E8EW64	Phb synthase subunits PhaC and PhaE	-
Clostridium acetireducens DSM 10703	A0A1E8EW93 AND A0A1E8EW64	Phb synthase subunits PhaC and PhaE	-

Synonyms

Synonyms	Comment	Organism
CLAOCE_21140	locus name	Clostridium acetireducens
CLAOCE_21150	locus name	Clostridium acetireducens
CLAOCE_21150/21140	locus name	Clostridium acetireducens
PHA synthase	-	Clostridium acetireducens
PhaEC	-	Clostridium acetireducens
PhbC	-	Clostridium acetireducens
type III PHA synthase	-	Clostridium acetireducens

General Information

General Information	Comment	Organism
evolution	the three genes phaJ (CLAOCE_21160) and phaEC (CLAOCE_21150 and CLAOCE_21140) are clustered in Clostridium acetireducens and encode a (R)-enoyl-CoA hydratase as well as a type III PHA synthase. The genes phaJ and phaEC from Clostridium acetireducens share high sequence similarity compared to genes encoding enzymes involved in PHB formation such as phaJ from Rhodospirillum rubrum or Haloferax mediterranei (Rru_A2964, HFX_1483) and phaEC from Synechocystis sp. PCC 6803	Clostridium acetireducens
physiological function	species that do contain type III PHA synthases (phaEC) are able to produce butyrate and possess in addition an (R)-enoyl-CoA hydratase (phaJ), suggesting that (R)-3-hydroxybutyryl-CoA is the starting point for PHB synthesis, whereas the (S)-isomer is used for butyrate formation	Clostridium acetireducens