Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Parker, J.B.; Walsh, C.T.
    Action and timing of BacC and BacD in the late stages of biosynthesis of the dipeptide antibiotic bacilysin (2013), Biochemistry, 52, 889-901.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.385 overexpression in Escherichia coli with an N-terminal His6tag Bacillus subtilis
6.3.2.49 overexpression in Escherichia coli with an N-terminal His6tag Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ Bacillus subtilis (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. The enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ Bacillus subtilis 168 (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. The enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin Bacillus subtilis the enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin Bacillus subtilis 168 the enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.385 Bacillus subtilis P39640
-
-
1.1.1.385 Bacillus subtilis 168 P39640
-
-
6.3.2.49 Bacillus subtilis P39641
-
-
6.3.2.49 Bacillus subtilis 168 P39641
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.385
-
 Bacillus subtilis
6.3.2.49
-
 Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. The enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin Bacillus subtilis (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. Exogenous enzyme is added into crude DELTAbacC extracts and dihydroanticapsin transformation is analyzed by LC-MS. Wild-type extract contains a significant amount of bacilysin and a minor amount of anticapsin, but the DELTA extract does not possess a detectable amount of either. However, the DELTAbacC extract contains accumulated amounts of masses corresponding to dihydroanticapsin and dihydrobacilysin, which contain the intact epoxide but possess a C7-hydroxyl moiety in place of the C7-ketone found in mature anticapsin and bacilysin. This finding is consistent with the assignment of the enzyme (BacC) as the dehydrogenase used to oxidize the C7-hydroxyl and strongly suggests that the enzyme (BacC) oxidation occurs sometime after the epoxidation of the cyclohexenol double bond. When the DELTAbacC extract is exposed to the enzyme (BacC) the C7-hydroxyl of hydroanticapsin is oxidized to generate anticapsin. The enzyme (BacC) has no oxidation activity on the dihydrobacilysin dipeptide Bacillus subtilis (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. The enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin Bacillus subtilis 168 (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
1.1.1.385 (1R,2S,5R,6S)-dihydroanticapsin + NAD+ (1R,2S,5R,6S)-dihydroanticapsin i.e. 3-[(1R,2S,5R,6S)-5-hydroxy-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine. Exogenous enzyme is added into crude DELTAbacC extracts and dihydroanticapsin transformation is analyzed by LC-MS. Wild-type extract contains a significant amount of bacilysin and a minor amount of anticapsin, but the DELTA extract does not possess a detectable amount of either. However, the DELTAbacC extract contains accumulated amounts of masses corresponding to dihydroanticapsin and dihydrobacilysin, which contain the intact epoxide but possess a C7-hydroxyl moiety in place of the C7-ketone found in mature anticapsin and bacilysin. This finding is consistent with the assignment of the enzyme (BacC) as the dehydrogenase used to oxidize the C7-hydroxyl and strongly suggests that the enzyme (BacC) oxidation occurs sometime after the epoxidation of the cyclohexenol double bond. When the DELTAbacC extract is exposed to the enzyme (BacC) the C7-hydroxyl of hydroanticapsin is oxidized to generate anticapsin. The enzyme (BacC) has no oxidation activity on the dihydrobacilysin dipeptide Bacillus subtilis 168 (1R,2S,6R)-anticapsin + NADH + H+ (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
1.1.1.385 (2S,4S,7R)-tetrahydrotyrosine + NAD+ (2S,4S,7R)-tetrahydrotyrosine is a surrogate substrate for the enzyme, because currently there is no route for purifying dihydroanticapsin (with the epoxide intact) from DELTAbacC strain extracts. No activity with (2S,4R,7R)-tetrahydrotyrosine Bacillus subtilis 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine + NADH + H+
-
 ?
1.1.1.385 (2S,4S,7R)-tetrahydrotyrosine + NAD+ (2S,4S,7R)-tetrahydrotyrosine is a surrogate substrate for the enzyme, because currently there is no route for purifying dihydroanticapsin (with the epoxide intact) from DELTAbacC strain extracts. No activity with (2S,4R,7R)-tetrahydrotyrosine Bacillus subtilis 168 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine + NADH + H+
-
 ?
6.3.2.49 ATP + L-alanine + (1R,2S,5R,6S)-dihydroanticapsin
-
 Bacillus subtilis ADP + phosphate + dihydrobacilysin
-
 ?
6.3.2.49 ATP + L-alanine + (1R,2S,5R,6S)-dihydroanticapsin
-
 Bacillus subtilis 168 ADP + phosphate + dihydrobacilysin
-
 ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin the enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin Bacillus subtilis ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin when the DELTAbacC extract is exposed to the enzyme (BacD) and the dihydroanticapsin:NAD+ 7-oxidoreductase (BacC) the C7-hydroxyl of hydroanticapsin is oxidized to generate anticapsin. (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine Bacillus subtilis ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin the enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin Bacillus subtilis 168 ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + (1R,2S,6R)-anticapsin when the DELTAbacC extract is exposed to the enzyme (BacD) and the dihydroanticapsin:NAD+ 7-oxidoreductase (BacC) the C7-hydroxyl of hydroanticapsin is oxidized to generate anticapsin. (1R,2S,6R)-anticapsin i.e. 3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine Bacillus subtilis 168 ADP + phosphate + bacilysin i.e. L-alanyl-3-[(1R,2S,6R)-5-oxo-7-oxabicyclo[4.1.0]hept-2-yl]-L-alanine ?
6.3.2.49 ATP + L-alanine + 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine is a surrogate substrate for the enzyme, because currently there is no route for purifying dihydroanticapsin (with the epoxide intact) from DELTAbacC strain extracts. A simultaneous incubation of the enzyme (ligase BacD) with oxidoreductase BaC, (2S,4S,7R)-tetrahydrotyrosine, NAD+, L-Ala, and ATP yields the dipeptide product L-Ala-2S,4S-cyclohexenoyl-Ala. This result indicates the BacD ligase can intercept the nascent 4S-cyclohexenonyl-Ala and ligate its N-terminus to L-Ala before its free amino group undergoes intramolecular addition Bacillus subtilis ADP + phosphate + L-alanyl-[3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine]
-
 ?
6.3.2.49 ATP + L-alanine + 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine 3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine is a surrogate substrate for the enzyme, because currently there is no route for purifying dihydroanticapsin (with the epoxide intact) from DELTAbacC strain extracts. A simultaneous incubation of the enzyme (ligase BacD) with oxidoreductase BaC, (2S,4S,7R)-tetrahydrotyrosine, NAD+, L-Ala, and ATP yields the dipeptide product L-Ala-2S,4S-cyclohexenoyl-Ala. This result indicates the BacD ligase can intercept the nascent 4S-cyclohexenonyl-Ala and ligate its N-terminus to L-Ala before its free amino group undergoes intramolecular addition Bacillus subtilis 168 ADP + phosphate + L-alanyl-[3-[(1S)-4-oxocyclohex-2-en-1-yl]-L-alanine]
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.385 BacC
-
 Bacillus subtilis
1.1.1.385 BSU37720 locus name Bacillus subtilis
1.1.1.385 ywfD
-
 Bacillus subtilis
6.3.2.49 alanine-anticapsin ligase
-
 Bacillus subtilis
6.3.2.49 BacD
-
 Bacillus subtilis
6.3.2.49 BSU37710 locus name Bacillus subtilis
6.3.2.49 L-Ala-L-aniticapsin ligase
-
 Bacillus subtilis
6.3.2.49 L-alanine-L-aniticapsin ligase
-
 Bacillus subtilis
6.3.2.49 YwfE
-
 Bacillus subtilis

General Information

EC Number General Information Comment Organism
1.1.1.385 malfunction dihydroanticapsin and dihydrobacilysin accumulate in extracts of a DELTAbacC strain and are processed to anticapsin and then bacilysin upon addition of BacC and BacD, respectively Bacillus subtilis
1.1.1.385 metabolism the enzyme is involved in the biosynthesis of the dipeptide antibiotic bacilysin Bacillus subtilis