Strain construction for enhanced production of spinosad via intergeneric protoplast fusion

Wang, Chao WangC.; Zhang, XiaoLin ZhangX.; Chen, Zhi ChenZ.; Wen, Ying; Yuan, Sheng

doi:10.1139/w09-064

Cited by 13 publications

(7 citation statements)

References 15 publications

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“…Wang et al used intergeneric protoplast fusion between S. spinosa and Streptomyces avermitilis to improve the starch-utilization problem (18). Huang et al proved that maltose-transport-related genes obviously promoted the growth of S. spinosa and resulted in improved spinosad yields (19).…”

Section: Discussionmentioning

confidence: 99%

A New Medium for Improving Spinosad Production by Saccharopolyspora spinosa

Yang¹,

Yuping

Yan³

et al. 2016

Jundishapur J Microbiol

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BackgroundSpinosad (a mixture of spinosyns A and D) is a unique natural pesticide produced by Saccharopolyspora spinosa. With regard to attempts to improve S. spinosa by classical mutagenesis, we propose that the bottleneck of screening out high-spinosad-production strains is probably caused by the fermentation media.ObjectivesThe current study aimed to identify a new medium to extensively investigate the potential of S. spinosa strains to produce spinosad.Materials and MethodsStatistical and regressive modeling methods were used to investigate the effects of the carbon source and to optimize the production media.ResultsThe spinosad production of S. spinosa Co121 increased 77.13%, from 310.44 ± 21.84 μg/mL in the initial fermentation medium (with glucose as the main carbon source) to 549.89 ± 38.59 μg/mL in a new optimized fermentation medium (98.0 g of mannitol, 43.0 g of cottonseed flour, 12.9 g of corn steep liquor, 0.5 g of KH2PO4, and 3.0 g of CaCO3 in 1 L of H2O; pH was adjusted to 7.0 before autoclaving). After screening 4,000 strains, an overall 3.33-fold increase was observed in spinosad titers, starting from the parental strain Co121 in the original fermentation medium and ending with the mutant strain J78 (1035 ± 34 μg/mL) in the optimized medium.ConclusionsThe optimized fermentation medium developed in this study can probably be used to improve spinosad production in screening industrial strains of S. spinosa.

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Section: Discussionmentioning

confidence: 99%

A New Medium for Improving Spinosad Production by Saccharopolyspora spinosa

Yang¹,

Yuping

Yan³

et al. 2016

Jundishapur J Microbiol

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“…Protoplast fusion technology has frequently been used for the genetic manipulation of fungi (for establishing heterokaryons) when few molecular genetic tools are available [24], [25]. By facilitating the discovery of new intermediates and hybrid antibiotics with beneficial properties [26], the large-scale enhancement of metabolite yields [27], and the construction of starch-utilizing strains [28], protoplast fusion has shown great potential for industrial applications. The present study, however, required that the genetic background of the recipient isolates be maintained when protoplast fusion was used to transfer hypovirulent dsRNA mycovirus to the recipients.…”

Section: Discussionmentioning

confidence: 99%

Transmission of Fusarium boothii Mycovirus via Protoplast Fusion Causes Hypovirulence in Other Phytopathogenic Fungi

Lee

Son

et al. 2011

PLoS ONE

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There is increasing concern regarding the use of fungicides to control plant diseases, whereby interest has increased in the biological control of phytopathogenic fungi by the application of hypovirulent mycoviruses as a possible alternative to fungicides. Transmission of hypovirulence-associated double-stranded RNA (dsRNA) viruses between mycelia, however, is prevented by the vegetative incompatibility barrier that often exists between different species or strains of filamentous fungi. We determined whether protoplast fusion could be used to transmit FgV1-DK21 virus, which is associated with hypovirulence on F. boothii (formerly F. graminearum strain DK21), to F. graminearum, F. asiaticum, F. oxysporum f. sp. lycopersici, and Cryphonectria parasitica. Relative to virus-free strains, the FgV1-DK21 recipient strains had reduced growth rates, altered pigmentation, and reduced virulence. These results indicate that protoplast fusion can be used to introduce FgV1-DK21 dsRNA into other Fusarium species and into C. parasitica and that FgV1-DK21 can be used as a hypovirulence factor and thus as a biological control agent.

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“…Previously reported methods to improve S. spinosa spinosad levels include classic mutagenesis and rational selection [27], protoplast regeneration [28], intergeneric protoplast fusion [29], and duplication of the genes involved in spinosad biosynthesis [30]. S. spinosa has a filamentous growth and used fatty acid as precursor for spinosad biosynthesis, certain amount oil need to be added during fermentation.…”

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confidence: 99%