Strain Improvement Program of Streptomyces roseosporus for Daptomycin Production

Rubio, Sara; Martínez-Cámara, Sonia; Fuente, J. L. de la; Rodríguez‐Sáiz, Marta; Barredo, José-Luis

doi:10.1007/978-1-0716-1358-0_20

Cited by 6 publications

(3 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To improve strains for industrial use, it is important to develop safe and stable strains. Up until now, the classical random mutagenesis method has been used as a mutagenesis method for the high production of secondary metabolites in microorganisms and many studies have reported successful strain improvement results (Dutta et al, 2019;Sha et al, 2020;Rubio et al, 2021;Zhang et al, 2018). In particular, Avermectin, an active ingredient for abamectin, which is currently used as a broad-spectrum nematicide, is also a compound produced by Streptomyces avermitilis.…”

Section: Discussionmentioning

confidence: 99%

Development of Spectinabilin Over-Producing Mutant Streptomyces sp. S-N87 having Nematicidal Activities

Kang

Kim

Moon

et al. 2023

Preprint

View full text Add to dashboard Cite

Spectinabilin (neoaureothin) is a rare nitrophenyl-substituted polyketide produced by some Streptomyces species. This compound is known to exhibit various biological activities such as anticancer, antibiotic, immunomodulatory, antimalarial and nematicidal effects. Despite being a valuable secondary metabolite for the development of novel drugs, the production yield of spectinabilin is < 200 mg/litre at the current level. To improve the yields of this promising compound without biosafety and regulation issues, this study conducted traditional mutagenesis. A total of 1,025 mutants were generated under high mortality conditions by NTG (N-methyl-N’-nitro-N-nitrosoguanidine) from parental strain Streptomyces sp. AN091965. One of the mutants, S-N87 showed up to about 10-fold spectinabilin productivity (354.8 ± 7.8 mg/L) compared to the parental strain Streptomyces sp. AN091965 (37.6 ± 5.6 mg/L) in flask culture conditions, representing the highest spectinabilin yield reported thus far. In addition, this strain showed a stable yield of 2.27 g/L even in a scaled-up environment (150 L tank fermentation), which suggests that the selected mutant is a genetically stable and robust strain. Further, the mutant Streptomyces sp. S-N87 that enhanced spectinabilin production stably showed a significant increase in nematicidal activities against pine wilt nematode compared to the parental strain. The present study is the first to develop a Streptomyces mutant that over-produces spectinabilin by traditional mutagenesis. Further studies such as whole-genome analysis and genetic modification are needed to provide a theoretical basis and insights into the polyketide synthase pathway of this talented mutant.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of Spectinabilin Over-Producing Mutant Streptomyces sp. S-N87 having Nematicidal Activities

Kang

Kim

Moon

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In contrast to the rational but partial reconfiguration of these regulatory networks, the random mutagenesis occurs through more overall alterations in the genome and is more straightforward as it is only based on the ultimate phenotypic outputs, for example, the productivity of natural products. It can be carried out with biological methods, such as insertion of mobile genetic elements transposons and random evolution by high resistance to antibiotics ( Xu et al, 2017 ; Luo et al, 2018 ; Yu et al, 2019a ), chemical agents, such as N -methyl- N′ -nitro- N -nitrosoguanidine (MNNG) ( Mo et al, 2013 ; Rubio et al, 2021 ) and more convenient physical measures mainly based on high-energy radiation from ultraviolet (UV) and the recently emerging atmospheric and room temperature plasma (ARTP) ( Mo et al, 2013 ; Yu et al, 2019b ; Ye et al, 2021 ), as well as the combinatory applications of above strategies ( Mo et al, 2013 ; Yan et al, 2021 ). However, the biggest challenge of the mutagenesis strategy is the high throughput but efficient screening of strains for hyper-production of the desired natural products.…”

Section: Introductionmentioning

confidence: 99%

Development of a native-locus dual reporter system for the efficient screening of the hyper-production of natural products in Streptomyces

Zhou

Zhao

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Streptomyces is renowned for its abundant production of bioactive secondary metabolites, but most of these natural products are produced in low yields. Traditional rational network refactoring is highly dependent on the comprehensive understanding of regulatory mechanisms and multiple manipulations of genome editing. Though random mutagenesis is fairly straightforward, it lacks a general and effective strategy for high throughput screening of the desired strains. Here in an antibiotic daptomycin producer S. roseosporus, we developed a dual-reporter system at the native locus of the daptomycin gene cluster. After elimination of three enzymes that potentially produce pigments by genome editing, a gene idgS encoding the indigoidine synthetase and a kanamycin resistant gene neo were integrated before and after the non-ribosomal peptidyl synthetase genes for daptomycin biosynthesis, respectively. After condition optimization of UV-induced mutagenesis, strains with hyper-resistance to kanamycin along with over-production of indigoidine were efficiently obtained after one round of mutagenesis and target screening based on the dual selection of the reporter system. Four mutant strains showed increased production of daptomycin from 1.4 to 6.4 folds, and significantly improved expression of the gene cluster. Our native-locus dual reporter system is efficient for targeting screening after random mutagenesis and would be widely applicable for the effective engineering of Streptomyces species and hyper-production of these invaluable natural products for pharmaceutical development.

show abstract

“…It has a calcium-dependent mechanism of action, which can destroy bacterial membranes. Given that it is effective against other antibiotic-resistant microorganisms, including vancomycin-resistant Staphylococcus aureus (VRSA) and methicillin-resistant Staphylococcus aureus (MRSA), daptomycin is in great demand and is widely used nowadays [14]. So far, more than 2000 actinomycetes have been documented in the literature, and about 600 are from the Streptomyces genus.…”

Section: Introductionmentioning

confidence: 99%

Secondary Metabolites from Streptomyces araujoniae S-03 Show Biocontrol Potential against Rhododendron Root Rot Caused by Phytophthora cinnamomi

Sun,

Wang,

et al. 2023

Forests

View full text Add to dashboard Cite

Phytophthora cinnamomi is a harmful microorganism that can infect Cinnamomum plants and cause the rotting of plant roots. It has been reported as infecting nearly 5000 types of plants worldwide, such as avocado trees, macadamia trees, and oak and chestnut trees, and is listed among the 10 most destructive oomycetes and the 100 most severely invasive species globally. A number of chemical agents have been applied in the control of phytophthora rot diseases because of their easy application and rapid effects. However, with the extensive use of chemical agents, P. cinnamomi has developed resistance. More importantly, it has damaged the ecological environment and affected human health. Given that biological control should be safe and effective, the screening of biocontrol strains with highly effective antagonistic effects is regarded as the primary means to control P. cinnamomi. In this study, a bacterial strain was isolated from the soil of healthy rhododendron, located near to diseased rhododendron plants, and identified as Streptomyces araujoniae, named S-03, which inhibited P. cinnamomi. The plate antagonism assay showed that S-03 could effectively bend the hyphae, reduce the number of branches, and even break them, destroying the integrity of the hyphal cell membrane to inhibit the growth of P. cinnamomi. Moreover, strain S-03 also could inhibit the activity of P. cinnamomi cell wall-degrading enzymes. To analyze the inhibitory mechanism of S-03, the effects of cell wall-degrading enzymes, secondary metabolites, and volatile substances produced by S-03 on P. cinnamomi were assessed. It was found that the fermentation broth of S-03 had a strong inhibitory effect on P. cinnamomi which means metabolites play a part in inhibition. The active substance of S-03 was initially separated and purified by ethyl acetate extraction and silica gel column chromatography and had the properties of small molecules, low polarity, and solubility in methanol. The biocontrol effect was detected in the host plants and indicated that S-03 could effectively protect rhododendron from P. cinnamomi infection. Overall, the present study findings provide compelling evidence that Streptomyces S-03 could be a biocontrol agent against plant diseases caused by P. cinnamomi.

show abstract

Strain Improvement Program of Streptomyces roseosporus for Daptomycin Production

Cited by 6 publications

References 13 publications

Development of Spectinabilin Over-Producing Mutant Streptomyces sp. S-N87 having Nematicidal Activities

Development of Spectinabilin Over-Producing Mutant Streptomyces sp. S-N87 having Nematicidal Activities

Development of a native-locus dual reporter system for the efficient screening of the hyper-production of natural products in Streptomyces

Secondary Metabolites from Streptomyces araujoniae S-03 Show Biocontrol Potential against Rhododendron Root Rot Caused by Phytophthora cinnamomi

Contact Info

Product

Resources

About