Streptomycin‐induced ribosome engineering complemented with fermentation optimization for enhanced production of 10‐membered enediynes tiancimycin‐A and tiancimycin‐D

Zhuang, Zhoukang; Jiang, Chengzhou; Zhang, Fan; Huang, Rong; Yi, Liwei; Huang, Yong; Yan, Xiaohui; Duan, Yanwen; Zhu, Xia

doi:10.1002/bit.26944

Cited by 30 publications

(35 citation statements)

References 29 publications

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“…A rif strain CB03234-R-16 and a str strain CB03234-S were obtained by treating the S. sp. CB03234 wild-type strain with rifampicin or streptomycin [48,49]. The CB03234-R-16 has an rpoB L422P mutation, while CB03234-S has an rpsL K43N mutation.…”

Section: Application Of Ribosome Engineering To Increase Antibiotimentioning

confidence: 99%

The Application of Ribosome Engineering to Natural Product Discovery and Yield Improvement in Streptomyces

2019

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Microbial natural product drug discovery and development has entered a new era, driven by microbial genomics and synthetic biology. Genome sequencing has revealed the vast potential to produce valuable secondary metabolites in bacteria and fungi. However, many of the biosynthetic gene clusters are silent under standard fermentation conditions. By rational screening for mutations in bacterial ribosomal proteins or RNA polymerases, ribosome engineering is a versatile approach to obtain mutants with improved titers for microbial product formation or new natural products through activating silent biosynthetic gene clusters. In this review, we discuss the mechanism of ribosome engineering and its application to natural product discovery and yield improvement in Streptomyces. Our analysis suggests that ribosome engineering is a rapid and cost-effective approach and could be adapted to speed up the discovery and development of natural product drug leads in the post-genomic era.

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Section: Application Of Ribosome Engineering To Increase Antibiotimentioning

confidence: 99%

The Application of Ribosome Engineering to Natural Product Discovery and Yield Improvement in Streptomyces

2019

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“…Therefore, introducing mutations conferring streptomycin resistance is widely used in bacteria and Streptomyces sp. for improving antibiotic and other metabolite production [22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Improved Neomycin Sulfate Potency in Streptomyces fradiae Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization

Zhang

Sun

et al. 2022

Microorganisms

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To improve the screening efficiency of high-yield neomycin sulfate (NM) Streptomyces fradiae strains after mutagenesis, a high-throughput screening method using streptomycin resistance prescreening (8 μg/mL) and a 24-deep well plates/microplate reader (trypan blue spectrophotometry) rescreening strategy was developed. Using this approach, we identified a high-producing NM mutant strain, Sf6-2, via six rounds of atmospheric and room temperature plasma (ARTP) mutagenesis and screening. The mutant displayed a NM potency of 7780 ± 110 U/mL and remarkably stable genetic properties over six generations. Furthermore, the key components (soluble starch, peptone, and (NH4)2SO4) affecting NM potency in fermentation medium were selected using Plackett-Burman and optimized by Box-Behnken designs. Finally, the NM potency of Sf6-2 was increased to 10,849 ± 141 U/mL at the optimal concentration of each factor (73.98 g/L, 9.23 g/L, and 5.99 g/L, respectively), and it exhibited about a 40% and 100% enhancement when compared with before optimization conditions and the wild-type strain, respectively. In this study, we provide a new S. fradiae NM production strategy and generate valuable insights for the breeding and screening of other microorganisms.

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“…Strain improvement of the TNM A producer Streptomyces sp. CB03234, in combination with medium optimization, has successfully improved TNM A production to exceed 20 mg/l (Liu et al, 2018 , 2020 ; Zhuang et al, 2019 ). Subsequently, the engineered S. sp.…”

Section: Introductionmentioning

confidence: 99%

Submerged fermentation of Streptomyces uncialis providing a biotechnology platform for uncialamycin biosynthesis, engineering, and production

Hindra

Yang

Luo

et al. 2021

Journal of Industrial Microbiology and Biotechnology

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Uncialamycin (UCM) belongs to the anthraquinone-fused subfamily of 10-membered enediyne natural products that exhibits an extraordinary cytotoxicity against a wide spectrum of human cancer cell lines. Antibody-drug conjugates (ADCs), utilizing synthetic analogues of UCM as payloads, are in preclinical development. UCM is exclusively produced by Streptomyces uncialis DCA2648 on solid agar medium with low titers (∼0.019mg/ L), limiting its supply by microbial fermentation and hampering its biosynthetic and engineering studies by in vivo pathway manipulation. Here we report cultivation conditions that enable genetic manipulation of UCM biosynthesis in vivo and allow UCM production, with improved titers, by submerged fermentation of the engineered S. uncialis strains. Specifically, the titer of UCM was improved nearly 58-fold to ∼1.1mg/ L through the combination of deletion of biosynthetic gene clusters encoding unrelated metabolites from the S. uncialis wild-type, chemical mutagenesis and manipulation of pathway-specific regulators to generate the engineered S. uncialis strains, and finally medium optimization of the latter for UCM production. Genetic manipulation of UCM biosynthesis was demonstrated by inactivating selected genes in the engineered S. uncialis strains, one of which afforded a mutant strain accumulating tiancimycin B, a common biosynthetic intermediate known for the anthraquinone-fused subfamily of enediyne natural products. These findings highlight a biotechnology platform for UCM biosynthesis, engineering, and production that should facilitate both its fundamental studies and translational applications.

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Streptomycin‐induced ribosome engineering complemented with fermentation optimization for enhanced production of 10‐membered enediynes tiancimycin‐A and tiancimycin‐D

Cited by 30 publications

References 29 publications

The Application of Ribosome Engineering to Natural Product Discovery and Yield Improvement in Streptomyces

The Application of Ribosome Engineering to Natural Product Discovery and Yield Improvement in Streptomyces

Improved Neomycin Sulfate Potency in Streptomyces fradiae Using Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Fermentation Medium Optimization

Submerged fermentation of Streptomyces uncialis providing a biotechnology platform for uncialamycin biosynthesis, engineering, and production

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