Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli as His-tagged protein | Bacillus subtilis |
gene rizB, two splicing variants, rizB-P1 and rizB-P2, transcriptional analysis and DNA and amino acid sequence determination and analysis, sequence comparison, recombinant overexpression of the His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | required. Mg2+ can be replaced with Mn2+ and Co2+. The use of Mn2+ or Co2+ decreases activity and the maximum length of peptide synthesized becomes shorter | Bacillus subtilis | |
Co2+ | can replace Mg2+, but with lower activity | Bacillus subtilis | |
Mg2+ | required. Mg2+ can be replaced with Mn2+ and Co2+. The use of Mn2+ or Co2+ decreases activity and the maximum length of peptide synthesized becomes shorter | Bacillus subtilis | |
Mg2+ | required, best metal cofactor, can be replaced by Mn2+ or Co2+ | Bacillus subtilis | |
Mn2+ | required. Mg2+ can be replaced with Mn2+ and Co2+. The use of Mn2+ or Co2+ decreases activity and the maximum length of peptide synthesized becomes shorter | Bacillus subtilis | |
Mn2+ | can replace Mg2+, but with lower activity | Bacillus subtilis | |
additional information | in the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+ | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | C0STU2 | - |
- |
Bacillus subtilis | C0STU2 | gene rizB, two variants P1 and P2 | - |
Bacillus subtilis NBRC3134 | C0STU2 | - |
- |
Bacillus subtilis NBRC3134 | C0STU2 | gene rizB, two variants P1 and P2 | - |
Purification (Comment) | Organism |
---|---|
- |
Bacillus subtilis |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Bacillus subtilis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2 | Baa: Leu, Ile, Met | Bacillus subtilis | |
3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa | Zaa: Arg, His, Gln, Asn or Phe | Bacillus subtilis | |
ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa | Yaa: Arg, Lys, His, Gln, Asn, Ala, Ser, Thr, Gly, Leu, Ile, Met, Phe or Trp | Bacillus subtilis | |
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n | Xaa: Val, Leu, or Met. n = 2-5 | Bacillus subtilis | |
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n | the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner, showing synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 ATP + 2 L-Val + Gly | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-Gly | - |
? | |
2 ATP + 2 L-Val + L-Ala | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Ala | - |
? | |
2 ATP + 2 L-Val + L-Ala | - |
Bacillus subtilis NBRC3134 | 2 ADP + 2 phosphate + L-Val-L-Val-L-Ala | - |
? | |
2 ATP + 2 L-Val + L-Arg | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Arg | - |
? | |
2 ATP + 2 L-Val + L-Arg | - |
Bacillus subtilis NBRC3134 | 2 ADP + 2 phosphate + L-Val-L-Val-L-Arg | - |
? | |
2 ATP + 2 L-Val + L-Asn | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Asn | - |
? | |
2 ATP + 2 L-Val + L-Gln | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Gln | - |
? | |
2 ATP + 2 L-Val + L-His | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-His | - |
? | |
2 ATP + 2 L-Val + L-Ile | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Ile | - |
? | |
2 ATP + 2 L-Val + L-Leu | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Leu | - |
? | |
2 ATP + 2 L-Val + L-Lys | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Lys | - |
? | |
2 ATP + 2 L-Val + L-Met | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Met | - |
? | |
2 ATP + 2 L-Val + L-Phe | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Phe | - |
? | |
2 ATP + 2 L-Val + L-Ser | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Ser | - |
? | |
2 ATP + 2 L-Val + L-Thr | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Thr | - |
? | |
2 ATP + 2 L-Val + L-Trp | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Trp | - |
? | |
2 ATP + 3 L-leucine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 2 ADP + 2 phosphate + L-Leu-L-Leu-L-Leu | - |
? | |
2 ATP + 3 L-methionine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 2 ADP + 2 phosphate + L-Met-L-Met-L-Met | - |
? | |
2 ATP + 3 L-valine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Val | - |
? | |
2 ATP + L-Val + 2 L-Leu | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Leu-L-Leu | - |
? | |
2 ATP + L-Val + 2 L-Met | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Met-L-Met-L-Met | - |
? | |
2 ATP + L-Val + L-Arg-L-Ser | no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate | Bacillus subtilis | 2 ADP + phosphate + L-Val-L-Arg-L-Ser | - |
? | |
2 ATP + L-Val + L-Val + L-Arg-L-Ser | no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate | Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Val-L-Arg-L-Ser | - |
? | |
2 ATP + Val + 2 L-Ile | - |
Bacillus subtilis | 2 ADP + 2 phosphate + L-Val-L-Ile-L-Ile | - |
? | |
3 ATP + 3 L-Val + L-Arg | - |
Bacillus subtilis | 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Arg | - |
? | |
3 ATP + 3 L-Val + L-Gln | - |
Bacillus subtilis | 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Gln | - |
? | |
3 ATP + 3 L-Val + L-His | - |
Bacillus subtilis | 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-His | - |
? | |
3 ATP + 3 L-Val + L-Phe | - |
Bacillus subtilis | 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Phe | - |
? | |
3 ATP + 4 L-leucine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 3 ADP + 3 phosphate + L-Leu-L-Leu-L-Leu-L-Leu | - |
? | |
3 ATP + 4 L-methionine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 3 ADP + 3 phosphate + L-Met-L-Met-L-Met-L-Met | - |
? | |
3 ATP + 4 L-valine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val | - |
? | |
4 ATP + 5 L-leucine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 4 ADP + 4 phosphate + L-Leu-L-Leu-L-Leu-L-Leu-L-Leu | - |
? | |
4 ATP + 5 L-methionine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 4 ADP + 4 phosphate + L-Met-L-Met-L-Met-L-Met-L-Met | - |
? | |
4 ATP + 5 L-valine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | 4 ADP + 4 phosphate + L-Val-L-Val-L-Val-L-Val-L-Val | - |
? | |
ATP + 2 L-leucine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | ADP + phosphate + L-Leu-L-Leu | - |
? | |
ATP + 2 L-methionine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | ADP + phosphate + L-Met-L-Met | - |
? | |
ATP + 2 L-valine | the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP | Bacillus subtilis | ADP + phosphate + L-Val-L-Val | - |
? | |
ATP + L-Val + L-Arg | - |
Bacillus subtilis | ADP + phosphate + L-Val-L-Arg | - |
? | |
ATP + L-Val + L-Arg-L-Ser | no homopeptides of Val-Val and Val-Val-Val are detected. The enzyme preferentially uses dipeptides as the C-terminal substrates | Bacillus subtilis | ADP + phosphate + L-Val-L-Arg-L-Ser + L-Val-L-Val-L-Arg-L-Ser | - |
? | |
ATP + L-Val + L-Arg-NHOH | - |
Bacillus subtilis | ADP + phosphate + L-Val-L-Arg-NHOH + L-Val-L-Val-L-Arg-NHOH | - |
? | |
ATP + L-Val + L-Phe-NHOH | - |
Bacillus subtilis | ADP + phosphate + L-Val-L-Phe-NHOH + L-Val-L-Val-L-Phe-NHOH | - |
? | |
additional information | dipeptides, tripeptides, tetrapeptides and pentapeptides are synthesized in the reaction mixtures containing Val, Leu, or Met as substrate. When Ile is used, the longest peptides are tetrapeptides. No peptides are detected when the other 15 amino acids are used as substrates. When Leu, Ile, or Met is used as substrate with Val, heteropeptides containing two residues of Leu, Ile, and Met are detected. No heteropeptides containing Tyr. Pro, Cys, Asp or Glu are detected. When other amino acids that are not usable for homopeptide synthesis are used as substrate, heteropeptides containing only one amino acid residue of these amino acids are detected. Substrate recognition at the N-terminus influences the resulting heteropeptide sequence. The enzyme does not participate in peptide-to-peptide ligation. Very low activity with D-amino acids | Bacillus subtilis | ? | - |
? | |
additional information | the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner. Synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. In the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus, mass spectrometric reaction analysis, overview | Bacillus subtilis | ? | - |
? | |
additional information | dipeptides, tripeptides, tetrapeptides and pentapeptides are synthesized in the reaction mixtures containing Val, Leu, or Met as substrate. When Ile is used, the longest peptides are tetrapeptides. No peptides are detected when the other 15 amino acids are used as substrates. When Leu, Ile, or Met is used as substrate with Val, heteropeptides containing two residues of Leu, Ile, and Met are detected. No heteropeptides containing Tyr. Pro, Cys, Asp or Glu are detected. When other amino acids that are not usable for homopeptide synthesis are used as substrate, heteropeptides containing only one amino acid residue of these amino acids are detected. Substrate recognition at the N-terminus influences the resulting heteropeptide sequence. The enzyme does not participate in peptide-to-peptide ligation. Very low activity with D-amino acids | Bacillus subtilis NBRC3134 | ? | - |
? | |
additional information | the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner. Synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. In the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus, mass spectrometric reaction analysis, overview | Bacillus subtilis NBRC3134 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
oligomer L-amino acid ligase | - |
Bacillus subtilis |
oligopeptide ligase | - |
Bacillus subtilis |
rizB | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bacillus subtilis |
40 | - |
- |
Bacillus subtilis |
40 | - |
recombinant enzyme | Bacillus subtilis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 45 | activity range, recombinant enzyme, profile overview | Bacillus subtilis |
30 | 45 | 30°C: about 35% of maximal activity, 45°C: about 80% of maximal activity | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Bacillus subtilis |
9 | - |
about, recombinant enzyme | Bacillus subtilis |
9 | 9.5 | - |
Bacillus subtilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.5 | 9.5 | activity range, recombinant enzyme, profile overview | Bacillus subtilis |
8 | 10 | pH 8.0: about 50% of maximal activity, significant decrease in activity above pH 10.0 | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | dependent on, cannot be replaced by GTP, CTP, or TTP | Bacillus subtilis |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme RizB shows high sequence homology to protein BL02410, UniProt ID Q65P25, from Bacillus licheniformis. In contrast to the other L-amino acid ligases, e.g. RizA and YwfE, RizB catalyzes the formation of oligomers from L-amino acids | Bacillus subtilis |
metabolism | the enzyme might be involved in the rhizocticin biosynthesis | Bacillus subtilis |
physiological function | the enzyme is involved in rhizocticin biosynthesis | Bacillus subtilis |