PXD040627

PXD040627 is an original dataset announced via ProteomeXchange.

Title	Expression of Blue Pigment Synthetase A from Streptomyces lavenduale Reveals Insights on the Effects of Refactoring Biosynthetic Megasynthases for Heterologous Expression in Escherichia coli
Description	High GC bacteria from the genus Streptomyces harbor expansive secondary metabolism. The expression of biosynthetic proteins and the characterization and identification of biological “parts” for synthetic biology purposes from such pathways are of interest. However, the high GC content of proteins from actinomycetes in addition to the large size and multi-domain architecture of many biosynthetic proteins (such as non-ribosomal peptide synthetases; NRPSs, and polyketide synthases; PKSs often called “megasynthases”) often presents issues with full-length translation and folding. Here we evaluate a non-ribosomal peptide synthetase (NRPS) from Streptomyces lavenduale, a multidomain “megasynthase” gene that comes from a high GC (72.5%) genome. While a preliminary step in revealing differences, to our knowledge this presents the first head-to-head comparison of codon-optimized sequences versus a native sequence of proteins of Streptomycete origin heterologously expressed in E. coli. We find that any disruption in co-translational folding from codon mismatch that compromises activity as measured by pigment titer is explainable via the formation of more inclusion bodies as opposed to compromising folding or posttranslational modification in the soluble fraction. This result supports that one could apply any refactoring strategies that improve soluble expression in E. coli without concern that the protein that reaches the soluble fraction has compromised folding.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_06:47:26.512.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD040627
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Christopher Petzold
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	O-phosphopantetheine-L-serine; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-03-06 09:54:38	ID requested
1	2023-06-23 08:39:54	announced
⏵ 2	2023-11-14 06:47:26	announced	2023-11-14: Updated project metadata.

10.6019/PXD040627;

submitter keyword: streptomyce, synthetic biology, codon optimization, refactoring,heterologous expression, natural products

Christopher Petzold
contact affiliation	Staff Scientist Biological Systems & Engineering Division Lawrence Berkeley National Laboratory Berkeley CA 94720
contact email	cjpetzold@lbl.gov
lab head
Christopher Petzold
contact affiliation	Lawrence Berkeley National Laboratory
contact email	cjpetzold@lbl.gov
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD040627
     1. Label: PRIDE project
     2. Name: Expression of Blue Pigment Synthetase A from Streptomyces lavenduale Reveals Insights on the Effects of Refactoring Biosynthetic Megasynthases for Heterologous Expression in Escherichia coli