Genome shuffling based on different types of ribosome engineering mutants for enhanced production of 10-membered enediyne tiancimycin-A

Liu, Huiming; Jiang, Chengzhou; Lin, Jing; Zhuang, Zhoukang; Kong, Wenping; Liu, Ling; Huang, Yong; Duan, Yanwen; Zhu, Xia

doi:10.1007/s00253-020-10583-2

Cited by 17 publications

(14 citation statements)

References 26 publications

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“…CB03234-G, a Genresistant mutant recently generated to overproduce tiancimycins. [22] The rimocidin high-yield producing mutants, Streptomyces rimosus M527-G37 and M527-GR7 with gentamicin resistance, have no mutations in either the 16S rRNA or the rplF gene. [36] The high-yield vitamin B12 producing bacterial strain KO-1261, derived from Shermanii ATCC 13673 with gentamicin resistance, shows no mutations in the ribosomal protein genes.…”

Section: Discussionmentioning

confidence: 99%

“…The optimized fermentation conditions may guide us for further pilot-scale production of YPMs in 30-or 150 L fermenters, which we have used for the pilot production of tiancimycins. [22]…”

Section: Discussionmentioning

confidence: 99%

“…[19][20][21] For example, the use of Rif or Gen alone, or in combination with genome shuffling, led to significant yield improvement of TNM A and the highest yield of TNM A reached to 40.6 ± 1.0 mg L −1 , about 135 folds of that in the wild-type strain (∼0.3 mg L −1 ). [22,23] Therefore, we hypothesize that ribosome engineering and fermentation optimization may be rapidly used to improve the yield of YPMs in M. yangpuensis DSM 45577. To our knowledge, there are few reports to use this strategy for yield improvement in Micromonospora species, despite that ribosome engineering has been used in various bacteria species, including Streptomyces, Bacillus, Pseudomonas, and Amycolatopsis.…”

Section: Introductionmentioning

confidence: 99%

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Yield improvement of enediyne yangpumicins in Micromonospora yangpuensis through ribosome engineering and fermentation optimization

Wang

Sun

et al. 2021

Biotechnology Journal

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Yangpumicins (YPMs), for example, YPM A, F, and G, are newly discovered enediynes from Micromonospora yangpuensis DSM 45577, which could be exploited as promising payloads of antibody-drug conjugates. However, the low yield of YPMs in the wildtype strain (∼1 mg L −1 ) significantly hampers their further drug development. In this study, a combined ribosome engineering and fermentation optimization strategy has been used for yield improvement of YPMs. One gentamicin-resistant M. yangpuensis DSM 45577 strain (MY-G-1) showed higher YPMs production (7.4 ± 1.0 mg L −1 ), while it exhibits delayed sporulation and slender mycelium under scanning electron microscopy. Whole genome re-sequencing of MY-G-1 reveals several deletion and single nucleotide polymorphism mutations, which were confirmed by PCR and DNA sequencing. Further Box-Behnken experiment and regression analysis determined that the optimal medium concentrations of soluble starch, D-mannitol, and pharmamedia for YPMs production in shaking flasks (10.0 ± 0.8 mg L −1 ). Finally, the total titer of YPM A/F/G in MY-G-1 reached to 15.0 ± 2.5 mg L −1 in 3 L fermenters, which was about 11-fold higher than the original titer of 1.3 ± 0.3 mg L −1 in wild-type strain. Our study may be instrumental to develop YPMs into a clinical anticancer drug, and inspire the use of these multifaceted strategies for yield improvement in Micromonospora species.

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Section: Discussionmentioning

confidence: 99%

“…The optimized fermentation conditions may guide us for further pilot-scale production of YPMs in 30-or 150 L fermenters, which we have used for the pilot production of tiancimycins. [22]…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Yield improvement of enediyne yangpumicins in Micromonospora yangpuensis through ribosome engineering and fermentation optimization

Wang

Sun

et al. 2021

Biotechnology Journal

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“…GS was carried out using the described methods with modifications (Wang et al, 2017;Liu et al, 2020). Spore of the starting strains were inoculated in 50mL Erlenmeyer flasks containing 30mL of seed medium and cultured at 28℃ and 220rpm for 2days.…”

Section: Genome Shufflingmentioning

confidence: 99%

Combination of atmospheric and room temperature plasma (ARTP) mutagenesis, genome shuffling and dimethyl sulfoxide (DMSO) feeding to improve FK506 production in Streptomyces tsukubaensis

et al. 2021

Biotechnol Lett

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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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Genome shuffling based on different types of ribosome engineering mutants for enhanced production of 10-membered enediyne tiancimycin-A

Cited by 17 publications

References 26 publications

Yield improvement of enediyne yangpumicins in Micromonospora yangpuensis through ribosome engineering and fermentation optimization

Yield improvement of enediyne yangpumicins in Micromonospora yangpuensis through ribosome engineering and fermentation optimization

Combination of atmospheric and room temperature plasma (ARTP) mutagenesis, genome shuffling and dimethyl sulfoxide (DMSO) feeding to improve FK506 production in Streptomyces tsukubaensis

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

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