Dataset Information

Enhanced protein secretion in reduced genome strains of Streptomyces lividans

ABSTRACT: S. lividans TK24 is a popular host for the production of small molecules and for the secretion of heterologous proteins. TK24 has a large genome with at least 29 secondary metabolite gene clusters that are non-essential for viability and undergo complex regulation. To optimize heterologous protein secretion, we previously constructed ten chassis strains that are devoid of several secondary metabolite gene clusters. Genome reduction was aimed at reducing carbon flow to secondary metabolites and pigmentation in the spent growth medium and improving colony morphology. Strains RG1.0-RG1.10 contain various deletion combinations of the blue actinorhodin cluster (act), the calcium-dependent antibiotic (cda), the undecylprodigiosin (red) and coelimycin A (cpk) clusters, the melanin cluster (mel), the matAB genes that affect mycelial aggregation and the non-essential sigma factor hrdD that controls the transcription of Act and Red regulatory proteins. Two derivative strains, RG1.5 and 1.9, showed a ~15% reduction in growth rate, >2-fold increase in the total mass yield of their native secretome and altered abundance of several specific proteins compared with TK24. Metabolomics and RNAseq analysis revealed that genome reduction led to rapid cessation of growth due to aminoacid depletion and caused both redox and cell envelope stresses, upregulation of the Sec-pathway components secDF and chaperones and a cell envelope two component regulator. RG1.9 maintained elevated heterologous secretion of mRFP and mTNFα by 12-70%. An integrated model is presented linking genome reduction and enhanced secretion. Importance: S. lividans TK24 encode 29 secondary metabolite gene clusters controlled with highly complex systems. This study established an important step toward understanding how secondary metabolite clusters can be manipulated to construct a surrogate TK24 platform with optimized metabolite funnelling. Using integrative multi-omics tools with protein secretion we provide an in-depth view of this fascinating complex process and its mechanistic regulation.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Streptomyces Lividans Tk24

SUBMITTER: Mohamed Belal Hamed Soliman

LAB HEAD: Molecular Bacteriology

PROVIDER: PXD040146 | Pride | 2024-01-26

REPOSITORIES: Pride

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Enhanced protein secretion in reduced genome strains of Streptomyces lividans.

Hamed Mohamed Belal MB Busche Tobias T Simoens Kenneth K Carpentier Sebastien S Kormanec Jan J Van Mellaert Lieve L Anné Jozef J Kalinowski Joern J Bernaerts Kristel K Karamanou Spyridoula S Economou Anastassios A

Microbial cell factories 20240105 1

<h4>Background</h4>S. lividans TK24 is a popular host for the production of small molecules and the secretion of heterologous protein. Within its large genome, twenty-nine non-essential clusters direct the biosynthesis of secondary metabolites. We had previously constructed ten chassis strains, carrying deletions in various combinations of specialized metabolites biosynthetic clusters, such as those of the blue actinorhodin (act), the calcium-dependent antibiotic (cda), the undecylprodigiosin (r ...[more]

PMID: 38183102

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