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Information on EC 1.2.1.75 - malonyl-CoA reductase (malonate semialdehyde-forming)
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1.2.1.75 malonyl-CoA reductase (malonate semialdehyde-forming)IUBMB Comments
Requires Mg2+. Catalyses the reduction of malonyl-CoA to malonate semialdehyde, a key step in the 3-hydroxypropanoate and the 3-hydroxypropanoate/4-hydroxybutanoate cycles, autotrophic CO2 fixation pathways found in some green non-sulfur phototrophic bacteria and some thermoacidophilic archaea, respectively [1,2]. The enzyme from Sulfolobus tokodaii has been purified, and found to contain one RNA molecule per two subunits . The enzyme from Chloroflexus aurantiacus is bifunctional, and also catalyses the next reaction in the pathway, EC 1.1.1.298 [3-hydroxypropionate dehydrogenase (NADP+)] .
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The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
bi-functional malonyl-CoA reductase, malonate semialdehyde reductase, malonate-semialdehyde dehydrogenase, malonyl CoA reductase (malonate semialdehyde-forming), malonyl-CoA reductase, MCR, More, MSAR, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
malonyl CoA reductase (malonate semialdehyde-forming)
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malonyl-CoA reductase
MCR
additional information
malonyl-CoA reductase
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MCR
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
malonate semialdehyde + CoA + NADP+ = malonyl-CoA + NADPH + H+
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malonate semialdehyde + CoA + NADP+ = malonyl-CoA + NADPH + H+
the protein acts as rigid template to press CoA and NADP into similar S-shaped, superimposable forms
malonate semialdehyde + CoA + NADP+ = malonyl-CoA + NADPH + H+
the protein acts as rigid template to press CoA and NADP into similar S-shaped, superimposable forms
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PATHWAY SOURCE
PATHWAYS
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SYSTEMATIC NAME
IUBMB Comments
malonate semialdehyde:NADP+ oxidoreductase (malonate semialdehyde-forming)
Requires Mg2+. Catalyses the reduction of malonyl-CoA to malonate semialdehyde, a key step in the 3-hydroxypropanoate and the 3-hydroxypropanoate/4-hydroxybutanoate cycles, autotrophic CO2 fixation pathways found in some green non-sulfur phototrophic bacteria and some thermoacidophilic archaea, respectively [1,2]. The enzyme from Sulfolobus tokodaii has been purified, and found to contain one RNA molecule per two subunits [3]. The enzyme from Chloroflexus aurantiacus is bifunctional, and also catalyses the next reaction in the pathway, EC 1.1.1.298 [3-hydroxypropionate dehydrogenase (NADP+)] [4].
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CAS REGISTRY NUMBER
COMMENTARY
429691-13-4
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
malonate semialdehyde + coenzyme A + NADP+
malonyl-CoA + NADPH + H+
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-
-
r
malonyl-CoA + NADPH + H+
malonate semialdehyde + NADP+ + CoA
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-
-
-
?
succinate semialdehyde + coenzyme A + NADP+
succinyl-CoA + NADPH + H+
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-
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r
succinyl-CoA + NADPH + H+
succinate semialdehyde + coenzyme A + NADP+
at 25% of the rate with malonyl-CoA
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r
malonate semialdehyde + NADPH + H+
3-hydroxypropionic acid + NADP+
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?
malonate semialdehyde + NADPH + H+
3-hydroxypropionic acid + NADP+
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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-
-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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-
-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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-
-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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-
-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
structural analysis reveals an unexpected reaction cycle in which NADP+ and CoA successively occupy identical binding sites, proposed reaction mechanism
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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-
-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
structural analysis reveals an unexpected reaction cycle in which NADP+ and CoA successively occupy identical binding sites, proposed reaction mechanism
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-
?
malonyl-CoA + NADPH + H+
malonate semialdehyde + coenzyme A + NADP+
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + coenzyme A + NADP+
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r
succinyl-CoA + NADPH + H+
succinic semialdehyde + CoA + NADP+
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-
?
succinyl-CoA + NADPH + H+
succinic semialdehyde + CoA + NADP+
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?
additional information
?
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enzyme additionally catalyzes the second reduction step of malonate semialdehyde + NADPH + H+ to 3-hydroxypropionate + NADP+. Reverse reaction starting with 3-hydroxypropionate does not require CoA and probably stops at malonate semialdehyde. No substrates are acetyl-CoA, propionyl-CoA, succinyl-CoA, or glyoxylate
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?
additional information
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the bifunctional enzyme shows malonate semialdehyde reduction activity, EC 1.1.1.298, and also malonyl-CoA reduction activity. The C-terminal subdomain MCR-C reduces malonyl-CoA to malonate semialdehyde, while the N-terminal subdomain MCR-N reduces malonate semialdehyde to 3-HP
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additional information
?
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the bifunctional malonyl-CoA reductase catalyzes the formation of malonate semialdehyde from malonyl-CoA, and the reduction of malonate semialdehyde to 3-hydroxypropionate, cf. EC 1.1.1.298, molecular mechanism of the conversion of malonyl-CoA to 3-HP in the bacterial 3-HP pathway, substrate binding docking simulations, overview
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additional information
?
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the bifunctional malonyl-CoA reductase catalyzes the formation of malonate semialdehyde from malonyl-CoA, and the reduction of malonate semialdehyde to 3-hydroxypropionate, cf. EC 1.1.1.298, molecular mechanism of the conversion of malonyl-CoA to 3-HP in the bacterial 3-HP pathway, substrate binding docking simulations, overview
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additional information
?
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the bifunctional malonyl-CoA reductase catalyzes the formation of malonate semialdehyde from malonyl-CoA, and the reduction of malonate semialdehyde to 3-hydroxypropionate, cf. EC 1.1.1.298, molecular mechanism of the conversion of malonyl-CoA to 3-HP in the bacterial 3-HP pathway, substrate binding docking simulations, overview
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
malonate semialdehyde + NADPH + H+
3-hydroxypropionic acid + NADP+
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-
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?
malonate semialdehyde + NADPH + H+
3-hydroxypropionic acid + NADP+
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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?
malonyl-CoA + NADPH + H+
malonate semialdehyde + CoA + NADP+
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?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADPH
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NADPH
the NADPH cofactor bound in MCR N-terminal domain is stabilized by hydrogen bonds with the side chains of Arg55, Arg59, Asp84, Asn151, Tyr744 and Lys195, and the main chains of Asn34, Leu35, Gly85, Asn111, Gly113 and Ile224, and by interaction with C-terminal resdiues by hydrogen bonds with the side chains of Ser88, Arg611, Arg612, Asp646, Tyr744 and Lys748, and the main chains of Ser588, Ala589, Ile591, Arg611, Arg612, Val647, Asn673 and Val776, cofactor binding site structure, overview
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.8
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
brenda
bifunctional aldehyde dehydrogenase plus alcohol dehydrogenase
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brenda
bifunctional enzyme
UniProt
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
the enzyme is most active in autotrophically grown Chloroflexus aurantiacus at 57°C
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
metabolism
physiological function
additional information
evolution
distribution of bifunctional MCR in bacteria and comparison with archaeal MCR and MSAR, overview
evolution
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distribution of bifunctional MCR in bacteria and comparison with archaeal MCR and MSAR, overview
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metabolism
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the enzyme participates in the 3-hydroxypropionate/4-hydroxybutyrate cycle, an autotrophic CO2 fixation pathway found in some thermoacidophilic archaea
metabolism
3-hydroxypropionic acid (3HP) production via MCR dependent pathway, overview. The bifunctional enzyme shows malonate semialdehyde reduction activity, EC 1.1.1.298, and also malonyl-CoA reduction activity
metabolism
enzymes involved in archaeal and bacterial 3-HP pathway and their structures, overview
metabolism
the bifunctional enzyme from Chloroflexus aurantiacus synthesizes 3-hydroxypropionate (3-HP) from acetate via malonyl-CoA in the malonyl-CoA reductase pathway, enzyme MCR shows malonyl-CoA reductase activity, EC 1.1.1.298, and converts malonyl-CoA to malonate semialdehyde and CoA using NADPH. The malonate semialdehyde is then reduced to 3-hydroxypropionic acid, overview
metabolism
the bifunctional enzyme from Chloroflexus aurantiacus synthesizes 3-hydroxypropionate (3-HP) from malonyl-CoA via the malonyl-CoA reductase pathway, it shows malonyl-CoA reductase activity and converts malonyl-CoA to malonate semialdehyde and CoA using NADPH. The malonate semialdehyde is then reduced to 3-hydroxypropionic acid, EC 1.1.1.298, overview
metabolism
the bifunctional enzyme from Chloroflexus aurantiacus synthesizes 3-hydroxypropionate (3-HP) from malonyl-CoA via the malonyl-CoA reductase pathway, it shows malonyl-CoA reductase activity and converts malonyl-CoA to malonate semialdehyde and CoA using NADPH. The malonate semialdehyde is then reduced to 3-hydroxypropionic acid, EC 1.1.1.298. 3HP can be produced from several intermediates, such as glycerol, malonyl-CoA, and beta-alanine. Among all these biosynthetic routes, the malonyl-CoA pathway has some distinct advantages, including a broad feedstock spectrum, thermodynamic feasibility, and redox neutrality. Comparison of the different metabolic routes for 3HP biosynthesis from glycerol or glucose, overview
metabolism
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enzymes involved in archaeal and bacterial 3-HP pathway and their structures, overview
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physiological function
the N-terminal region of MCR (MCR-N, amino acids 1-549) and the C-terminal region of MCR (MCR-C, amino acids 550-1219) are functionally distinct. Malonyl-CoA is reduced into free intermediate malonate semialdehyde with NADPH by the MCR-C fragment, and further reduced to 3-hydroxypropionate by the MCR-N fragment, the initial reduction of malonyl-CoA being rate limiting. The TGXXXG(A)X(1-2)G and YXXXK motifs are important for enzyme activities of both MCR-N and MCR-C fragments, and the enzyme activity increases when MCR is separated into two individual fragments. The MCR-C fragment has higher affinity for malonyl-CoA and 4-times higher Kcat/Km value than MCR
physiological function
the bifunctional enzyme from Chloroflexus aurantiacus synthesizes 3-hydroxypropionate (3-HP) from malonyl-CoA via the malonyl-CoA reductase pathway, it shows malonyl-CoA reductase activity and converts malonyl-CoA to malonate semialdehyde and CoA using NADPH. The malonate semialdehyde is then reduced to 3-hydroxypropionic acid, EC 1.1.1.298, overview
additional information
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Tyr191 is the catalytic residue, active site structure, substrate binding mode, overview. Structure comparison with the archaeal MCR from Sulfurisphaera tokodaii (StMCR)
additional information
Tyr191 is the catalytic residue, active site structure, substrate binding mode, overview. Structure comparison with the archaeal MCR from Sulfurisphaera tokodaii (StMCR)
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MCR_METS5
Metallosphaera sedula (strain ATCC 51363 / DSM 5348 / JCM 9185 / NBRC 15509 / TH2)
357
0
39255
Swiss-Prot
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MCR_SULTO
Sulfurisphaera tokodaii (strain DSM 16993 / JCM 10545 / NBRC 100140 / 7)
359
0
39560
Swiss-Prot
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A0A841L016_9SPHN
1226
0
130070
TrEMBL
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A0A1A7BFR5_9SPHN
1230
0
132356
TrEMBL
-
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
39000
45000
39000
or dimer, 4 * 39000, calculated, 4 * 45000, SDS-PAGE, enzyme contains bound RNA
39000
or tetramer, 2 * 39000, calculated, 2 * 45000, SDS-PAGE, enzyme contains bound RNA
45000
or dimer, 4 * 39000, calculated, 4 * 45000, SDS-PAGE, enzyme contains bound RNA
45000
or tetramer, 2 * 39000, calculated, 2 * 45000, SDS-PAGE, enzyme contains bound RNA
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
homodimer
homotetramer
tetramer
additional information
dimer
or tetramer, 2 * 39000, calculated, 2 * 45000, SDS-PAGE, enzyme contains bound RNA
tetramer
or dimer, 4 * 39000, calculated, 4 * 45000, SDS-PAGE, enzyme contains bound RNA
additional information
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olecular architecture of the full-length MCR, modeling, overview
additional information
olecular architecture of the full-length MCR, modeling, overview
additional information
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olecular architecture of the full-length MCR, modeling, overview
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additional information
enzyme in its native state is associated with small RNA
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallization trials of the full-length MCR fail, thus production of the N-terminal domain (MCRND, Met1-Pro567) and the C-terminal domain (MCRCD, Gly568-Val1230) separately, X-ray diffraction structure determination and analysis at resolutions of 2.20 and 1.80 A, respectively, by a single-wavelength anomalous dispersion (SAD) method
N-terminal domain, hanging drop vapor diffusion method, using 10% (w/v) PEG3350, 200 mM lithium sulfate, 100 mM Tris-HCl, pH 8.0. C-terminal domain, hanging drop vapor diffusion method, using 1.6 M lithium sulfate and 100 mM HEPES, pH 8.0
malonyl-CoA reductase in the substrate-free state at 2.05 A resolution and in complex with NADP+ at 1.9 A resolution and in complex with CoA at 2.4 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
N940V/K1106W/S1114R
I798A
the mutant of the C-terminal domain shows less than 30% activity as compared to the wild type enzyme
K195A
the mutant of the N-terminal domain is almost completely inactive as compared to the wild type enzyme
K748A
the mutant of the C-terminal domain is almost completely inactive as compared to the wild type enzyme
K802A
the mutant of the C-terminal domain shows about 95% activity as compared to the wild type enzyme
K926A
the mutant of the C-terminal domain shows about 90% activity as compared to the wild type enzyme
M662A
the mutant of the C-terminal domain is almost completely inactive as compared to the wild type enzyme
N740A
the mutant of the C-terminal domain shows about 15% activity as compared to the wild type enzyme
N805A
the mutant of the C-terminal domain shows less than 60% activity as compared to the wild type enzyme
R1166A
the mutant of the C-terminal domain shows less than 20% activity as compared to the wild type enzyme
R188A
the mutant of the N-terminal domain shows less than 10% activity as compared to the wild type enzyme
R741A
the mutant of the C-terminal domain shows less than 5% activity as compared to the wild type enzyme
R780A
the mutant of the C-terminal domain is almost completely inactive as compared to the wild type enzyme
R794A
the mutant of the C-terminal domain is almost completely inactive as compared to the wild type enzyme
S726A
the mutant of the C-terminal domain is almost completely inactive as compared to the wild type enzyme
T178A
the mutant of the N-terminal domain is almost completely inactive as compared to the wild type enzyme
Y185A
the mutant of the N-terminal domain shows less than 20% activity as compared to the wild type enzyme
Y191A
the mutant of the N-terminal domain shows less than 10% activity as compared to the wild type enzyme
Y738A
the mutant of the C-terminal domain shows about 10% activity as compared to the wild type enzyme
Y744A
the mutant of the C-terminal domain shows less than 5% activity as compared to the wild type enzyme
synthesis
additional information
N940V/K1106W/S1114R
site-directed mutagenesis, mutant N940V/K1106W/S1114R improves the catalytic efficiency by 14.2fold over the wild-type
N940V/K1106W/S1114R
site-directed mutagenesis, the mutant shows increased enzyme activity compared to wild-type enzyme
N940V/K1106W/S1114R
the mutations improve the catalytic efficiency by 14.2fold over the wild type
N940V/K1106W/S1114R
the mutations increase the activity of C-terminal of the enzyme by 5.54fold as compared to the wild type
synthesis
heterologous expression of the malonyl-CoA dependent pathway genes malonyl-CoA reductase and malonate semialdehyde reductase enables Synechococcus elongatus to synthesize 3-hydroxypropionic acid to a final titer of 665 mg/l
synthesis
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heterologous expression of the malonyl-CoA dependent pathway genes malonyl-CoA reductase and malonate semialdehyde reductase enables Synechococcus elongatus to synthesize 3-hydroxypropionic acid to a final titer of 665 mg/l
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additional information
3-hydroxypropionate (3HP) is an attractive platform chemical, serving as a precursor to a variety of commodity chemicals like acrylate and acrylamide, as well as a monomer of a biodegradable plastic. It can be used to establish a sustainable way to produce these commercially important chemicals and materials, fermentative production of 3HP is widely investigated in recent years. Reconstruction of the malonyl-CoA pathway in Escherichia coli employing acetyl-CoA carboxylase (ACC) for the conversion of acetyl-CoA into malonyl-CoA, which is converted into 3HP with a two-step reduction catalyzed by malonyl-CoA reductase (MCR) that converts malonyl-CoA to malonate semialdehyde and CoA, malonate semialdehyde is then reduced to 3-hydroxypropionic acid (EC 1.1.1.298). Redirection of carbon flux toward 3HP biosynthesis by metabolic engineering e.g. through manipulation of various regulation factors controlling central carbon metabolism, such as CsrB, SgrS and ArcA, or through inhibition of the activity of 3-oxoacyl-ACP synthase I and II with the antibiotic cerulenin to suppress fatty acids biosynthesis, or through improving catalysis of key enzymes, enhancing cofactor and energy supply, and promoting catalytic efficiency of MCR. Compared to Escherichia coli, Saccharomyces cerevisiae is the better host
additional information
engineering of type II methanotroph Methylosinus trichosporium strain OB3b for 3-hydroxypropionic acid (3HP) production by reconstructing malonyl-CoA pathway through heterologous expression of Chloroflexus aurantiacus malonyl-CoA reductase (MCR), a bifunctional enzyme. Engineering of the supply of malonyl-CoA precursors by overexpressing endogenous acetyl-CoA carboxylase (ACC), substantially enhancing the production of 3HP. Overexpression of biotin protein ligase (BPL) and malic enzyme (NADP+-ME) leads to 22.7% and 34.5% increase, respectively, in 3HP titer in ACC-overexpressing cells. Also, the acetyl-CoA carboxylation bypass route is reconstructed to improve 3HP productivity. Coexpression of methylmalonyl-CoA carboxyltransferase (MMC) of Propionibacterium freudenreichii and phosphoenolpyruvate carboxylase (PEPC), which provides the MMC precursor, further improves the 3HP titer. The highest 3HP production of 49 mg/l in the OB3b-MCRMP strain overexpressing MCR, MMC and PEPC results in a 2.4fold improvement of titer compared with that in the only MCR-overexpressing strain. 60.59 mg/l of 3HP are obatined in 42 h using the OB3b-MCRMP strain through bioreactor operation, with a 6.36fold increase of volumetric productivity compared than that in the flask cultures
additional information
enhancing 3-hydroxypropionic acid production in combination with sugar supply engineering by cell surface-display and metabolic engineering of Schizosaccharomyces pombe. 3-HP production from glucose and cellobiose via the malonyl-CoA pathway, the mcr gene, encoding the bifunctional malonyl-CoA reductase of Chloroflexus aurantiacus, is dissected into two functionally distinct fragments, and the activities of the encoded protein are balanced. The MCR-C fragment reduces malonyl-CoA to malonate semialdehyde, while the MCR-N fragment reduces malonate semialdehyde to 3-HP. To increase the cellular supply of malonyl-CoA and acetyl-CoA, genes encoding endogenous aldehyde dehydrogenase, acetyl-CoA synthase from Salmonella enterica, and endogenous pantothenate kinase are introduced. The resulting strain produces 3-HP at 1.0 g/l from a culture starting at a glucose concentration of 50 g/l. We also engineered the sugar supply by displaying beta-glucosidase (BGL) on the yeast cell surface. When grown on 50 g/l cellobiose, the beta-glucosidase-displaying strain consumes cellobiose efficiently and produces 3-HP at 3.5 g/l. Under fed-batch conditions starting from cellobiose, this strain produces 3-HP at up to 11.4 g/l, corresponding to a yield of 11.2%
additional information
for the efficient conversion of acetate to 3-hydroxypropionate(3-HP), heterologous mcr (encoding malonyl-CoA reductase) mutant N940V/K1106W/S1114R from Chloroflexus aurantiacus is initially introduced into Escherichia coli. Then, the acetate assimilating pathway and glyoxylate shunt pathway are activated by overexpressing acs (encoding acetyl-CoA synthetase) and deleting iclR (encoding the glyoxylate shunt pathway repressor). Because a key precursor malonyl-CoA is also consumed for fatty acid synthesis, carbon flux to fatty acid synthesis is inhibited by adding cerulenin, which dramatically improves 3-HP production. Method evaluation and optimization, overview
additional information
production of 3-hydroxypropionate using a novel malonyl-CoA-mediated biosynthetic pathway in genetically engineered Escherichia coli strain. Heterologously coexpressing the mutant of malonyl-CoA reductase (MCR) from Chloroflexus aurantiacus and malonyl-CoA synthetase (MatB) from Rhodopseudomonas palustris in the Escherichia coli C43 (DE3) strain. To further enhance the production of 3-HP, native transhydrogenase (PntAB) and NAD kinase (YfjB) genes are expressed to increase the NADPH supply in Escherichia coli. The final genetically modified strain SGN78 shows a significant improvement in malonate utilization and produced 1.20 g/l of 3-HP in the flask culture. Identification of suitable malonate transporters in Rhodobacter capsulatus and Sinorhizobium meliloti, and coexpression of transporter MatB in Escherichia coli. The enzyme activity increases when the N-terminal and C-terminal regions of MCR are separated by fusing a flexible linker (GGGGS) between the two enzymatic units. Optimization of fermentation conditions and improvement of NADPH supply increase 3-HP production rate
additional information
the malonyl-CoA reductase pathway involving the enzyme is successfully constructed in Saccharomyces cerevisiae, developments in 3-hydroxypropionate production using yeast as an industrial host, method, overview. Requirement of improving the supply of the cofactor NADPH due to high expense of NADPH
additional information
the crystallographic data indicate how to construct a bispecific cofactor binding site and to engineer a malonyl-CoA into methylmalonyl-CoA reductase for polyester building block production
additional information