MUTAGENESIS OF HIS-21; HIS-78; HIS-188 AND HIS-192
Catalyzes the cleavage of beta-carotene at its central double bond (15,15') to yield two molecules of all-trans-retinal. Exhibits the highest activity for beta-carotene, followed by beta-cryptoxanthin, beta-apo-4'-carotenal, alpha-carotene, and gamma-carotene in decreasing order, but has no activity on beta-apo-8'-carotenal, beta-apo-12'-carotenal, lutein, zeaxanthin, or lycopene, suggesting that the presence of one unsubstituted beta-ionone ring in a substrate with a molecular weight greater than C35 seems to be essential for enzyme activity.
The kcat is 4.5-fold higher with beta-carotene than with beta-cryptoxanthin as substrate.
Optimum temperature is 40 degrees Celsius. Above this temperature, the enzyme activity decreases significantly, exhibiting 66% of the maximum activity at 50 degrees Celsius. Below 40 degrees Celsius, the enzyme activity decreases with decreasing temperature, exhibiting 15% of the maximum activity at 25 degrees Celsius. The half-lives of the enzyme at 35, 40, 45, 50, and 55 degrees Celsius are 17.6, 15.0, 12.5, 6.1, and 1.5 hours, respectively.
Could be used for high retinal production.
Belongs to the Brp/Blh beta-carotene diooxygenase family.
HOMO-OLIGOMERIC FLAVIN CONTAINING CYS DECARBOXYLASE FAMILY
PHOSPHOPANTOTHENOYLCYSTEINE DECARBOXYLASE
Homo-oligomeric flavin-containing Cys decarboxylases, HFCD
Cysteine/Cysteine sulfinic acid decarboxylase
Catalyzes the decarboxylation of L-cysteine to cysteamine and of 3-sulfino-L-alanine (cysteine sulfinic acid) to hypotaurine (PubMed:17420004, Ref.2, Ref.3, PubMed:29796005). Also catalyzes the decarboxylation of various amino acids such as L-lysine, L-glutamate, L-asparaginate and L-proline (Ref.2). In vitro, shows highest activity with L-cysteine as substrate (Ref.2).
Slightly stimulated in the presence of 1 mM Mg(2+).
kcat is 4.57 min(-1) with L-cysteine as substrate (Ref.2). kcat is 45.8 min(-1) with L-cysteine sulfinic acid as substrate (Ref.3).
Optimum temperature is 35 degrees Celsius.
In the N-terminal section; belongs to the HFCD (homo-oligomeric flavin containing Cys decarboxylase) superfamily.
In the C-terminal section; belongs to the PPC synthetase family.
Three species of the Botryosphaeriales overlap on five unrelated trees in China, with a novel species.
MUTAGENESIS OF 152-LYS-GLU-153; SER-268; CYS-291 AND HIS-409
Displays some glucuronoyl esterase activity in vitro, since it is able to hydrolyze methyl 4-O-methyl-D-glucopyranosyluronate, allyl D-glucuronate, benzyl D-glucuronate and D-glucuronic acid methyl ester. However, esters of glucuronic acid are probably not its biological substrate, as they are not present in the marine environment. Can also hydrolyze a range of other esters, including p-nitrophenyl acetate. More likely biologically-relevant substrates for MZ0003 and other marine bacterial CE15s are algal cell wall polysaccharides, as these would be readily available in this environment and could be used as energy sources.
Does not require metal ions for activity.
Is inhibited by PMSF and by NaF in vitro, which is consistent with the catalytic nucleophile being a serine.
kcat is 8.9 sec(-1) with p-nitrophenyl acetate as substrate (in the absence of NaCl). kcat is 12.78 sec(-1) with p-nitrophenyl acetate as substrate (in the presence of 1M NaCl).
0.71 mM for p-nitrophenyl acetate (in the absence of NaCl)
0.97 mM for p-nitrophenyl acetate (in the presence of 1M NaCl)
Optimum pH is 8.0. Shows more than 60% of its maximum activity in the pH range of 7.0-9.5. 20% of the activity is retained at low pH range and at pH 10.5, while no catalytic activity is found at pH 5.0 or 10.5.
Optimum temperature is 35 degrees Celsius with p-nitrophenyl acetate as substrate. Above 35 degrees Celsius, activity significantly decreases, while 30% of the maximum activity is recorded at 10 degrees Celsius.
Belongs to the carbohydrate esterase 15 (CE15) family.
X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEXES WITH ANALOGS SUBSTRATE
X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEXES WITH ANALOGS SUBSTRATES
Tetrahydrodipicolinate-N-succinyltransferase, N-terminal domain
2,3,4,5-TETRAHYDROPYRIDINE-2,6-DICARBOXYLATE N-SUCCINYLTRANSFERASE
Tetrahydrodipicolinate N-succinyltransferase
Inhibited by p-(chloromercuri)benzenesulfonic acid and cobalt.
Belongs to the transferase hexapeptide repeat family.
Was originally thought to originate from Mycobacterium bovis (PubMed:8880935, PubMed:9012664, PubMed:9671504, PubMed:11910040). However, there is now convincing evidence that this is incorrect (PubMed:19394346). The source is unknown, but the sequence similarity suggests the protein is of enterobacterial origin.
Glutamate Dehydrogenase, chain A, domain 3
Leucine Dehydrogenase, chain A, domain 1
NADP-SPECIFIC GLUTAMATE DEHYDROGENASE 1-RELATED
Aminoacid dehydrogenase-like, N-terminal domain
Catalyzes the reversible oxidative deamination of glutamate to a-ketoglutarate and ammonia.
Belongs to the Glu/Leu/Phe/Val dehydrogenases family.
This sequence originates from an organism contaminating a Chlorella sorokiniana culture, probably a bacterium.
Uncultured organism feruloyl coenzyme A synthetase (fcs) gene.
Glutathione synthetase ATP-binding domain-like
Catalyzes the formation of feruloyl-CoA, ADP and phosphate from ferulate, CoA and ATP.
Optimum temperature is 37 degrees Celsius.
Catalyzes the activation of ferulate via CoA-thioesterification, a first enzymatic step for conversion of the lignin-derived ferulate into high value chemicals, such as vanillin and bioplastics.
In the N-terminal section; belongs to the acetate CoA ligase alpha subunit family.
In the C-terminal section; belongs to the acetate CoA ligase beta subunit family.
Saline water microbial communities from Organic Lake, Antarctica - #987.
NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]
Precursor of a pore-forming protein involved in defense against bacteriophages (By similarity). Cleavage of this precursor by its dedicated protease releases the active moiety (gasdermin bGSDM, N-terminus) which inserts into membranes, forming pores and triggering cell death (PubMed:35025633). Expression of bGSDM and the neighboring protease gene (Ga0307981_100051430) is highly toxic in E.coli (PubMed:35025633).
Pore-forming protein that causes membrane permeabilization via a pyroptosis-like activity. This is the active form which makes ring-like pores with an interior pore diameter of 130-190 Angstroms, when integrated in liposomes.
The full-length protein before cleavage is inactive: intramolecular interactions between the N-terminal domain and the C-terminal region mediate autoinhibition. The pyroptosis-like-inducing activity is carried by the released N-terminal domain (Gasdermin bGSDM, N-terminus).
Forms large, probably homooligomeric ring-shaped pores when inserted in membranes.
Upon proteolysis the protein forms membrane-associated puncta.
The N-terminus has marked structural similarity to the mammalian gasdermin N-terminal domain. The C-terminal region wraps around the twisted beta sheet core, probably stabilizing the inactive state.
Belongs to the bacterial gasdermin family.
Cleavage; by protease Ga0307981_100051430
X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS) IN COMPLEX WITH ZINC IONS
AMIDOHYDRO-REL DOMAIN-CONTAINING PROTEIN
Composite domain of metallo-dependent hydrolases
Belongs to the metallo-dependent hydrolases superfamily. ATZ/TRZ family.
Nitrilases catalyze the mild hydrolytic conversion of organonitriles directly to the corresponding carboxylic acids. Catalyzes the production of aryllactic acid derivatives. Mediates the hydrolysis of cyanohydrin to (S)-phenyllactic acid.
Belongs to the carbon-nitrogen hydrolase superfamily. Nitrilase family.
Nitrilases catalyze the mild hydrolytic conversion of organonitriles directly to the corresponding carboxylic acids.
Involved in the ornithine fermentation pathway. Catalyzes the oxidative deamination of (2R,4S)-2,4-diaminopentanoate ((2R,4S)-DAP) to yield 2-amino-4-ketopentanoate (AKP). Although Ord is more efficient with (2R,4S)-DAP, the diastereoisomer (2R,4R)-DAP can also be metabolized. Ord is specific for NAD as hydrogen acceptor.
Allosterically activated by (2R,4S)-DAP. Negatively regulated by AKP, and by both acetyl-CoA and D-alanine.
kcat is 34 sec(-1) with NAD as hydrogen acceptor. kcat is 5 sec(-1) with NADP as hydrogen acceptor.
2-amino-4-ketopentanoate thiolase alpha subunit
Involved in the ornithine fermentation pathway. Catalyzes the thiolytic cleavage of 2-amino-4-ketopentanoate (AKP) with coenzyme A (CoA) to form acetyl-CoA and alanine. It is strictly specific for AKP.
kcat is 3.5 sec(-1) for CoA as substrate. kcat is 3.4 sec(-1) for AKP as substrate.
Tryptophan synthase beta subunit-like PLP-dependent enzymes
2-amino-4-ketopentanoate thiolase beta subunit
Belongs to the threonine synthase family.
X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 36-293
X-RAY CRYSTALLOGRAPHY (1.10 ANGSTROMS) OF 36-293 OF MUTANTS ALA-165 AND ILE-243/CYS-238/CYS-283/GLY-127
Catalyzes the hydrolysis of cutin, a polyester that forms the structure of plant cuticle (PubMed:22194294). Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters), with a preference for short-chain substrates (C4 substrate at most) (PubMed:22194294, PubMed:24593046). Cannot hydrolyze olive oil (PubMed:22194294). Is also able to degrade poly(ethylene terephthalate), the most abundant polyester plastic in the world (PubMed:22194294, PubMed:32269349). Can also depolymerize poly(epsilon-caprolactone) (PCL), a synthetic aliphatic biodegradable polyester (PubMed:22194294).
Is inhibited in vitro by diethyl pNP-phosphate (E600), but not by EDTA.
kcat is 232 sec(-1) with pNP-butanoate as substrate (at 30 degrees Celsius and pH 8.0). kcat is 343 sec(-1) with pNP-butanoate as substrate (at 50 degrees Celsius and pH 8.0). kcat is 318 sec(-1) with pNP-butanoate as substrate (at 70 degrees Celsius and pH 8.0).
0.22 mM for pNP-butanoate (at 30 degrees Celsius and pH 8.0)
0.21 mM for pNP-butanoate (at 50 degrees Celsius and pH 8.0)
0.24 mM for pNP-butanoate (at 70 degrees Celsius and pH 8.0)
Optimum pH is 8.5 with pNP-butyrate as substrate. Shows 70% of the maximal activity at pH 7.0 and pH 9.5.
Optimum temperature is 50 degrees Celsius with pNP-butanoate as substrate (PubMed:22194294, PubMed:24593046). Optimum temperature using PET as substrate is superior to 70 degrees Celsius (PubMed:22194294). Shows 70% of the maximal activity at 30 and 70 degrees Celsius with pNP-butyrate as substrate. Has half-lives of 5 hours at 50 degrees Celsius, 80 minutes at 60 degrees Celsius, 40 minutes at 70 degrees Celsius and 7 minutes at 80 degrees Celsius (PubMed:22194294). Is thermostable, with a determined melting temperature (Tm) of 84.7 degrees Celsius (PubMed:32269349).
The disulfide bond between Cys-275 and Cys-292 contributes not only to the thermodynamic stability but also to the kinetic stability of the enzyme.
The engineered enzyme with mutations Ile-243/Cys-238/Cys-283/Gly-127 can hydrolyze PET in a highly efficient way, ultimately achieving, in less than 10 hours, a minimum of 90 per cent PET depolymerization into monomers, with a productivity of 16.7 grams of terephthalate per liter per hour. It was also shown that biologically recycled PET produced from enzymatically depolymerized PET waste exhibits the same properties as petrochemical PET, before being processed into bottles, thereby contributing towards the concept of a circular PET economy.
The gene encoding this enzyme comes from a leaf-branch compost metagenome. The source organism remains to be identified. However, it is presumably a thermophilic bacterium.
Belongs to the AB hydrolase superfamily.
Increased PET-depolymerization activity.
Although this is said to be a human protein, it is not encoded in the human genome. It is highly similar to microbial proteins and belongs to a family of microbial proteins.
PHYTANOYL-COA DIOXYGENASE DOMAIN CONTAINING 1
Alpha-ketoglutarate/Fe(II)-dependent dioxygenase PhnY
Involved in the degradation of the organophosphonate 2-aminoethylphosphonic acid (2-AEP) (Probable). Catalyzes the hydroxylation of 2-aminoethylphosphonic acid to yield (2-amino-1-hydroxyethyl)phosphonic acid (PubMed:22564006).
X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEX WITH IRON IONS AND SUBSTRATE
X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH IRON IONS AND SUBSTRATE
MUTAGENESIS OF TYR-24; GLU-27; HIS-34; HIS-58; ASP-59; HIS-62; HIS-80; HIS-104 AND ASP-161
2-amino-1-hydroxyethylphosphonate dioxygenase (glycine-forming)
Involved in the degradation of the organophosphonate 2-aminoethylphosphonic acid (2-AEP) (Probable). Catalyzes the cleavage of the carbon-phosphorus bond of (2-amino-1-hydroxyethyl)phosphonic acid to yield glycine and phosphate through an oxidative mechanism (PubMed:22564006, PubMed:24198335, PubMed:24706911). It reacts stereospecifically with the R-enantiomer of (2-amino-1-hydroxyethyl)phosphonic acid and is also able to use (R,R)-2-amino-1-hydroxypropylphosphonate as substrate (PubMed:24706911).
Binds 2 iron ions per subunit. During catalysis, PhnZ uses a mixed-valent Fe(2+)/Fe(3+) cofactor.
kcat is 11 min(-1) for (R)-2-amino-1-hydroxyethylphosphonic acid as substrate. kcat is 7 min(-1) for (R,R)-2-amino-1-hydroxypropylphosphonic acid as substrate.
0.16 mM for (R,R)-2-amino-1-hydroxypropylphosphonic acid
0.17 mM for (R)-2-amino-1-hydroxyethylphosphonic acid
Important for stereospecificity with the R-enantiomer
Slight decrease of the catalytic efficiency and of the affinity for (R)-2-amino-1-hydroxyethylphosphonic acid.
Slight decrease of the catalytic efficiency and of the affinity for (R)-2-amino-1-hydroxyethylphosphonic acid. 3-fold decrease of the Fe-enzyme molar ratios.
4-fold decrease of the catalytic efficiency and 3-fold decrease of the affinity for (R)-2-amino-1-hydroxyethylphosphonic acid. 6-fold decrease of the Fe-enzyme molar ratios.
6-fold decrease of the catalytic efficiency and 2.5-fold decrease of the affinity for (R)-2-amino-1-hydroxyethylphosphonic acid.
3-fold decrease of the catalytic efficiency and 3.5-fold decrease of the affinity for (R)-2-amino-1-hydroxyethylphosphonic acid. 2.5-fold decrease of the Fe-enzyme molar ratios.
P-loop containing nucleotide triphosphate hydrolases
Winged helix-like DNA-binding domain superfamily/Winged helix DNA-binding domain
LEUCINE-RICH REPEAT AND IQ DOMAIN-CONTAINING PROTEIN 1
LEUCINE-RICH REPEAT AND IQ DOMAIN-CONTAINING PROTEIN 1-RELATED
P-loop containing nucleoside triphosphate hydrolases
A dedicated protease for substrate gasdermin bGSDM; cleaves the bGSDM precursor, releasing the pore-forming moiety, which integrates into the membrane and triggers cell death. Probably involved in defense against bacteriophages. Expression of bGSDM and this neighboring protease is highly toxic in E.coli.
STRUCTURE BY ELECTRON MICROSCOPY (2.93 ANGSTROMS) OF 19-250 IN COVALENT COMPLEX WITH RETINAL
MUTAGENESIS OF LYS-58; TRP-59; TYR-96; GLU-143; SER-180; TYR-209; TYR-224 AND GLU-246
Rhodopsin 7-helix transmembrane proteins
BACTERIORHODOPSIN /OPSIN, NOPA (EUROFUNG)-RELATED
Family A G protein-coupled receptor-like
Homopentamer (PubMed:34226545, PubMed:32647827). GPR protomers assemble into a pentamer around a central pore with a C5 symmetry axis (PubMed:34226545).
Contains one covalently linked retinal chromophore per subunit.
Belongs to the archaeal/bacterial/fungal opsin family.
CNF1/YfiH-like putative cysteine hydrolases
S-methyl-5'-thioadenosine phosphorylase RL5
Purine nucleoside enzyme that catalyzes the phosphorolysis of adenosine and inosine nucleosides, yielding D-ribose 1-phosphate and the respective free bases, adenine and hypoxanthine (By similarity). Also catalyzes the phosphorolysis of S-methyl-5'-thioadenosine into adenine and S-methyl-5-thio-alpha-D-ribose 1-phosphate (By similarity). Also has adenosine deaminase activity (By similarity). Also acts as a multicopper oxidase able to oxidize a wide variety of polyphenols and related compounds in vitro (PubMed:16740638). Displays substrate preference as follows: syringaldazine > 2,6-dimethoxyphenol > veratryl alcohol > guaiacol > tetramethylbenzidine > 4-methoxybenzyl alcohol > 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) >> phenol red > 1-hydroxybenzotriazole (PubMed:16740638). Cannot use 3,4-dimetoxybenzyl alcohol and violuric acid as substrates (PubMed:16740638). As this enzyme is derived from a rumen microbial community, it may have a role in the digestion of complex plant materials such as ryegrass lignin (PubMed:16740638).
kcat is 18 sec(-1) with 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) as substrate. kcat is 660 sec(-1) with syringaldazine as substrate. kcat is 1175 sec(-1) with 2,6-dimethoxyphenol as substrate (at pH 4.5 and 40 degrees Celsius).
26 uM for 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (at pH 4.5 and 40 degrees Celsius)
0.43 uM for syringaldazine (at pH 4.5 and 40 degrees Celsius)
0.45 uM for 2,6-dimethoxyphenol (at pH 4.5 and 40 degrees Celsius)
Optimum pH is 4.0-5.0. Activity is very high over a broad pH range from 4.0 to 9.0. Exhibits more than 70% activity at pH 3.5 and 9.5.
Optimum temperature is about 60 degrees Celsius. Is fully stable at this temperature.
Belongs to the purine nucleoside phosphorylase YfiH/LACC1 family.
Loss of 1.0 of Cu/mol of protein. Loss of catalytic activity.
No effect on Cu content. No effect on catalytic activity.
Loss of 1.0 of Cu/mol of protein. No effect on secondary structure. Loss of catalytic activity.
Binds only 1.0 of Cu/mol of protein. Causes major change in secondary structure. Displays about 10% of wild-type catalytic activity.
Binds only 1.0 of Cu/mol of protein. Causes major change in secondary structure. Loss of catalytic activity.
X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH FAD ALONE AND IN COMPLEX WITH SUBSTRATE AND INHIBITOR
D-Amino Acid Oxidase, subunit A, domain 2
An FAD-requiring monooxygenase active on tetracycline antibiotic and some of its derivatives, which leads to their inactivation (PubMed:26097034). Expression in E.coli confers high resistance to tetracycline and oxytetracycline, does not confer resistance to minocycline or tigecycline. Degrades tetracycline and oxytetracycline; the reaction requires NADPH (PubMed:26097034). Degrades and confers resistance to chlortetracycline (PubMed:28481346).
Binds 1 FAD per subunit; it is seen bound in two conformations.
kcat for tetracycline is 4.3 min(-1), kcat for chlortetracycline is 3.5 min(-1).
Consists of an N-terminal FAD-binding domain with a Rossman fold, a substrate-binding domain and a C-terminal helix that bridges the 2 domains close to the antibiotic-binding site. This last helix is flexible, is not found in TetX of Bacteroides species, and may contribute to their different substrate specificities (PubMed:28481346). Binds chlortetracycline in a different orientation (180 degrees rotated on its long axis) compared to tetracycline substrate binding to TetX; binding of chlortetracycline alters conformation FAD and the last helix (PubMed:28481346).
Tetracycline antibiotics bind to the ribosomal acceptor site (A-site), preventing binding of the aminoacyl-tRNA to the A-site. The hydrophilic side of tetracycline makes many hydrogen-bonding interactions with oxygen atoms of the ribosome's phosphate backbone.
Belongs to the aromatic-ring hydroxylase family.
An FAD-requiring monooxygenase active on tetracycline antibiotic and some of its derivatives, which leads to their inactivation (PubMed:26097034). Expression in E.coli confers high resistance to oxytetracycline, slightly less resistance to tetracycline, moderate resistance to minocycline but no resistance to tigecycline. Degrades tetracycline and oxytetracycline; the reaction requires NADPH (PubMed:26097034). Degrades and confers resistance to chlortetracycline (PubMed:28481346).
Consists of an N-terminal FAD-binding domain with a Rossman fold, a substrate-binding domain and a C-terminal helix that bridges the 2 domains close to the antibiotic-binding site. This last helix is flexible, is not found in TetX of Bacteroides species, and may contribute to their different substrate specificities.
Isolated from an agricultural soil sample.