Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms
Abstract Background Natamycin is a natural antibiotic with broad-spectrum antifungal activity, widely used in food preservation, medicine, and biological control. However, the relatively low biosynthetic capacity of producing strains limits further industrialization and broader applications of natam...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-04-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-025-02696-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850258513901649920 |
|---|---|
| author | Liang Wang Wen Xiao Ting Qiu Hongjian Zhang Jianhua Zhang Xusheng Chen |
| author_facet | Liang Wang Wen Xiao Ting Qiu Hongjian Zhang Jianhua Zhang Xusheng Chen |
| author_sort | Liang Wang |
| collection | DOAJ |
| description | Abstract Background Natamycin is a natural antibiotic with broad-spectrum antifungal activity, widely used in food preservation, medicine, and biological control. However, the relatively low biosynthetic capacity of producing strains limits further industrialization and broader applications of natamycin. Due to the complexity of cellular metabolism, evolutionary engineering is required for developing strains with enhanced natamycin biosynthetic capacity. Results Here, protoplast fusion combined with phosphate tolerance screening was employed for the first time to enhance natamycin production of Streptomyces gilvosporeus. A high-yielding strain, GR-2, was obtained, with natamycin production twice that of the original strain. Transcriptomic analysis revealed that the natamycin biosynthetic gene cluster and several primary metabolic pathways were significantly upregulated in GR-2, likely contributing to its high production performance. Further experiments, including amino acid addition and reverse engineering, confirmed that branched-chain amino acid, nitrogen, and phosphate metabolism play crucial roles in promoting natamycin production. Silencing of the phosphate metabolism transcriptional regulators PhoP and PhoR led to a decreased expression of natamycin biosynthetic genes and significantly reduced natamycin production, highlighting the key role of these regulators in S. gilvosporeus. Based on omics data, co-expression of phoP and phoR in GR-2 resulted in the engineered strain GR2-P3, which exhibited a 25% increase in natamycin production in shake flasks. In a 5 L fermenter, GR2-P3 achieved a natamycin production of 12.2 ± 0.6 g·L⁻¹, the highest yield reported for S. gilvosporeus to date. Conclusions Our findings suggest that the high production performance of GR-2 is primarily due to the upregulation of the natamycin biosynthetic gene cluster and genes related to precursor supply. Increasing the intracellular supply of valine and glutamate significantly enhanced natamycin production. Additionally, the natamycin biosynthetic gene cluster is likely positively regulated by PhoP and PhoR. Our work presents a novel strategy for strain screening and evolution to improve natamycin production and identifies novel molecular targets for metabolic engineering. |
| format | Article |
| id | doaj-art-56499f8ec84a4bdf84ca306ae9353bfd |
| institution | OA Journals |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-56499f8ec84a4bdf84ca306ae9353bfd2025-08-20T01:56:09ZengBMCMicrobial Cell Factories1475-28592025-04-0124111610.1186/s12934-025-02696-yEnhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanismsLiang Wang0Wen Xiao1Ting Qiu2Hongjian Zhang3Jianhua Zhang4Xusheng Chen5Key Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, School of Biotechnology, Ministry of Education, Jiangnan UniversityAbstract Background Natamycin is a natural antibiotic with broad-spectrum antifungal activity, widely used in food preservation, medicine, and biological control. However, the relatively low biosynthetic capacity of producing strains limits further industrialization and broader applications of natamycin. Due to the complexity of cellular metabolism, evolutionary engineering is required for developing strains with enhanced natamycin biosynthetic capacity. Results Here, protoplast fusion combined with phosphate tolerance screening was employed for the first time to enhance natamycin production of Streptomyces gilvosporeus. A high-yielding strain, GR-2, was obtained, with natamycin production twice that of the original strain. Transcriptomic analysis revealed that the natamycin biosynthetic gene cluster and several primary metabolic pathways were significantly upregulated in GR-2, likely contributing to its high production performance. Further experiments, including amino acid addition and reverse engineering, confirmed that branched-chain amino acid, nitrogen, and phosphate metabolism play crucial roles in promoting natamycin production. Silencing of the phosphate metabolism transcriptional regulators PhoP and PhoR led to a decreased expression of natamycin biosynthetic genes and significantly reduced natamycin production, highlighting the key role of these regulators in S. gilvosporeus. Based on omics data, co-expression of phoP and phoR in GR-2 resulted in the engineered strain GR2-P3, which exhibited a 25% increase in natamycin production in shake flasks. In a 5 L fermenter, GR2-P3 achieved a natamycin production of 12.2 ± 0.6 g·L⁻¹, the highest yield reported for S. gilvosporeus to date. Conclusions Our findings suggest that the high production performance of GR-2 is primarily due to the upregulation of the natamycin biosynthetic gene cluster and genes related to precursor supply. Increasing the intracellular supply of valine and glutamate significantly enhanced natamycin production. Additionally, the natamycin biosynthetic gene cluster is likely positively regulated by PhoP and PhoR. Our work presents a novel strategy for strain screening and evolution to improve natamycin production and identifies novel molecular targets for metabolic engineering.https://doi.org/10.1186/s12934-025-02696-yNatamycinStreptomyces gilvosporeusPhosphate-tolerance screeningComparative transcriptomicsTranscriptional regulation |
| spellingShingle | Liang Wang Wen Xiao Ting Qiu Hongjian Zhang Jianhua Zhang Xusheng Chen Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms Microbial Cell Factories Natamycin Streptomyces gilvosporeus Phosphate-tolerance screening Comparative transcriptomics Transcriptional regulation |
| title | Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms |
| title_full | Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms |
| title_fullStr | Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms |
| title_full_unstemmed | Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms |
| title_short | Enhanced Natamycin production in Streptomyces gilvosporeus through phosphate tolerance screening and transcriptome-based analysis of high-yielding mechanisms |
| title_sort | enhanced natamycin production in streptomyces gilvosporeus through phosphate tolerance screening and transcriptome based analysis of high yielding mechanisms |
| topic | Natamycin Streptomyces gilvosporeus Phosphate-tolerance screening Comparative transcriptomics Transcriptional regulation |
| url | https://doi.org/10.1186/s12934-025-02696-y |
| work_keys_str_mv | AT liangwang enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms AT wenxiao enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms AT tingqiu enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms AT hongjianzhang enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms AT jianhuazhang enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms AT xushengchen enhancednatamycinproductioninstreptomycesgilvosporeusthroughphosphatetolerancescreeningandtranscriptomebasedanalysisofhighyieldingmechanisms |