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1.14.13.25: methane monooxygenase (soluble)

This is an abbreviated version!
For detailed information about methane monooxygenase (soluble), go to the full flat file.

Word Map on EC 1.14.13.25

Reaction

methane
+
NAD(P)H
 +
H+
 +
O2
=
methanol
 +
NAD(P)+
 +
H2O

Synonyms

chcA, cytoplasmic methane monooxygenase, methane hydroxylase, methane mono-oxygenase, methane monooxygenase, methane monooxygenase hydroxylase, MmMmoC, MMO, MMO Bath, MMOB, MmoC, MMOH, MMOR, oxygenase, methane mono-, particulate methane monooxygenase, pMMO, sMMO, soluble methane monooxygenase, soluble methane monooxygenase hydroxylase

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.13 With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
                1.14.13.25 methane monooxygenase (soluble)

Cloned

Cloned on EC 1.14.13.25 - methane monooxygenase (soluble)

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic analysis, quantitative expression analysis
DNA and amino acid sequence determination and analysis. The sMMO gene cluster consists of the structural genes mmoXYBZDC, the regulatory genes mmoG and mmoR and another ORF orf1. These sMMO genes are transcribed as a single unit from a s54-dependent promoter located upstream of mmoX
DNA sequence analysis
-
expressed in Escherichia coli
expression in Escherichia coli
expression in Escherichia coli, determination of complete sMMO DNA gene sequence, phylogenetic analysis
-
expression of component protein B of sMMO as glutathione-S-transferase fusion protein in Escherichia coli
-
expression of His-tagged triple mutant G10A/G13Q/G16A and G13Q mutant in Escherichia coli
-
gene mmoX, DNA and amino acid sequence determination and analysis
gene mmoX, quantitative RT-PCR enzyme expression analysis
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
genes mmoB encoding MMOB and mmoC encoding MMOR, recombinant expression of subunits in Escherichia coli BL21(DE3), and gene mmoD encoding MMOD is recombinantly expressed as His-MBP-tagged protein in Escherichia coli strain Rosetta (DE3)
-
mutants are expressed in sMMO-negative cells of Methylosinus trichosporium
-
overexpression of an additional protein component D of sMMO encoded by orfY as thioredoxin-fusion protein with His-Tag in Escherichia coli, protein component D is termed MMOD
-
overexpression of enzyme component MMOB in Escherichia coli
-
phylogenetic analysis
-
plasmids encoding wild-type MMOB and W77F and K15C MMOB are transformed into Escherichia coli BL21(DE3) chemically competent cells
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
quantitative expression analysis of subunits alpha, beta and gamma
-
quantitative RT-PCR expression analysis of genes pmoA, mmoX, and mbnA
-
recombinant enzyme expression in Escherichia coli strain BL21(DE3)
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
substantial improvements to the system for mutagenesis of soluble methane monooxygenase and expression of recombinant enzymes in the methanotroph Methylosinus trichosporium OB3b expression system. This system can be utilised to make a number of new mutants and to engineer soluble methane monooxygenase to increase its catalytic precision with a specific substrate whilst increasing activity by up to 6fold
P27353; P27355; P27354; P27356; Q53563; Q53562
transcription of sMMO is repressed at Cu2+ concentration above 0.00086 mM per g dry cell weight
-