Difference between revisions of "Part:BBa K302010"
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− | <partinfo> | + | <partinfo>BBa_K1152010 short</partinfo> |
− | ' | + | svp encodes for a 4'-Phosphopanthetheinyl-transferase (PPTase) which transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thus activating these enzymes. |
− | + | ||
− | + | ||
+ | [[File:Heidelberg2013_PPtase_mechanism.png|280px|thumb|'''Figure 1:''' General mechanism of activation the activation of NRP Synthetases from apo- to holo-form.]] | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
− | + | Svp is a PPTase often associated with activating the indigoidine synthetase bpsA [Takahashi, 2007]. | |
<!-- --> | <!-- --> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
− | <partinfo> | + | <partinfo>BBa_K1152010 SequenceAndFeatures</partinfo> |
+ | |||
+ | ===References=== | ||
+ | * Takahashi, H., Kumagai, T., Kitani, K., Mori, M., Matoba, Y., & Sugiyama, M. (2007). Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis *, 282(12), 9073–9081. doi:10.1074/jbc.M611319200 | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
− | <partinfo> | + | <partinfo>BBa_K1152010 parameters</partinfo> |
<!-- --> | <!-- --> | ||
+ | |||
+ | ===Information contributed by City of London UK (2021)=== | ||
+ | Part information is collated here to help future users of the BioBrick registry. | ||
+ | |||
+ | Metadata: | ||
+ | *'''Group:''' City of London UK 2021 | ||
+ | *'''Author:''' Janusan Jeyananthan | ||
+ | *'''Summary:''' Added information collated from existing scientific studies | ||
+ | |||
+ | ---- | ||
+ | |||
+ | Kinetics: | ||
+ | |||
+ | kcat is 86 min(-1) with S.glaucescens apo-ACP TcmM as the substrate and 11 min(-1) with S.mobaraensis apo-PCP BlmI as the substrate. | ||
+ | |||
+ | For Streptomyces mobaraensis apo-PCP BlmI1: KM=3.9 µM | ||
+ | For Streptomyces glaucescens apo-ACP TcmM: KM=3.1 µM | ||
+ | |||
+ | <ref>“Svp - 4’-Phosphopantetheinyl Transferase Svp - Streptomyces Mobaraensis - Svp Gene & Protein” n.d. Uniprot. Accessed 12 August, 2021 https://www.uniprot.org/uniprot/Q9F0Q6 </ref> | ||
+ | |||
+ | [[File:BBa_K302010.png|x200px|center]] | ||
+ | |||
+ | ===References=== | ||
+ | |||
+ | |||
+ | ===Information contributed by NFLS_Nanjing (2022)=== | ||
+ | Part information is collated here to help future users of the BioBrick registry. | ||
+ | |||
+ | Metadata: | ||
+ | *'''Group:''' NFLS_Nanjing (2022) | ||
+ | *'''Author:''' Yuming Xue | ||
+ | *'''Summary:''' Added information collated from existing scientific studies | ||
+ | |||
+ | Biology and usage: | ||
+ | |||
+ | 1. The gene bpsA is expressed in C. glutamicum and carries strong fluxes toward L-glutamate, a precursor of indigoidine. [1] | ||
+ | |||
+ | 2. The gene bpsA catalyzes indigoidine formation from two molecules of glutamine in an ATP-dependent manner.[2] | ||
+ | |||
+ | Experimental approach: | ||
+ | |||
+ | 1. Integrated sfp into the yeast chromosomal δ-sequences. | ||
+ | |||
+ | 2. Codon-optimize Te bpsA gene for expression in S. cerevisiae and genomically integrated into locus ARS1014a under control of the TDH3 promoter and ADH1 terminator using a previously reported, cloning free Cas9 toolkit. | ||
+ | |||
+ | 3. Use the conventional lithium acetate method to perform transformations. | ||
+ | |||
+ | 4. Use 200 ng pCut_1014a and 500 ng of linear Donor DNA with 500 bp homology to the integration locus ARS1014a.[3] | ||
+ | |||
+ | ===References=== | ||
+ | |||
+ | [1] High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes | ||
+ | |||
+ | [2] Genome-scale metabolic rewiring improves titersrates and yields of the non-native product indigoidine at scale | ||
+ | |||
+ | [3] Production efciency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae |
Latest revision as of 15:07, 18 September 2022
Svp 4'-Phosphopanthetheinyl-transferase
svp encodes for a 4'-Phosphopanthetheinyl-transferase (PPTase) which transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thus activating these enzymes.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 173
Illegal NgoMIV site found at 413
Illegal NgoMIV site found at 663
Illegal AgeI site found at 573 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 366
References
- Takahashi, H., Kumagai, T., Kitani, K., Mori, M., Matoba, Y., & Sugiyama, M. (2007). Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis *, 282(12), 9073–9081. doi:10.1074/jbc.M611319200
Information contributed by City of London UK (2021)
Part information is collated here to help future users of the BioBrick registry.
Metadata:
- Group: City of London UK 2021
- Author: Janusan Jeyananthan
- Summary: Added information collated from existing scientific studies
Kinetics:
kcat is 86 min(-1) with S.glaucescens apo-ACP TcmM as the substrate and 11 min(-1) with S.mobaraensis apo-PCP BlmI as the substrate.
For Streptomyces mobaraensis apo-PCP BlmI1: KM=3.9 µM For Streptomyces glaucescens apo-ACP TcmM: KM=3.1 µM
References
Information contributed by NFLS_Nanjing (2022)
Part information is collated here to help future users of the BioBrick registry.
Metadata:
- Group: NFLS_Nanjing (2022)
- Author: Yuming Xue
- Summary: Added information collated from existing scientific studies
Biology and usage:
1. The gene bpsA is expressed in C. glutamicum and carries strong fluxes toward L-glutamate, a precursor of indigoidine. [1]
2. The gene bpsA catalyzes indigoidine formation from two molecules of glutamine in an ATP-dependent manner.[2]
Experimental approach:
1. Integrated sfp into the yeast chromosomal δ-sequences.
2. Codon-optimize Te bpsA gene for expression in S. cerevisiae and genomically integrated into locus ARS1014a under control of the TDH3 promoter and ADH1 terminator using a previously reported, cloning free Cas9 toolkit.
3. Use the conventional lithium acetate method to perform transformations.
4. Use 200 ng pCut_1014a and 500 ng of linear Donor DNA with 500 bp homology to the integration locus ARS1014a.[3]
References
[1] High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes
[2] Genome-scale metabolic rewiring improves titersrates and yields of the non-native product indigoidine at scale
[3] Production efciency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae- ↑ “Svp - 4’-Phosphopantetheinyl Transferase Svp - Streptomyces Mobaraensis - Svp Gene & Protein” n.d. Uniprot. Accessed 12 August, 2021 https://www.uniprot.org/uniprot/Q9F0Q6