1.One-step fermentation for producing simvastatin via RNAi silencing of lovF gene in Aspergillus terreus.
Na LI ; Meihong LI ; Zhen YU ; Jun ZHOU ; Jun HUANG
Chinese Journal of Biotechnology 2016;32(4):478-486
Simvastatin is one of the important prescription drugs for hypercholesterolemia. Monacolin J is a key intermediate during simvastatin synthesis, and also an intermediate of lovastatin biosynthesis. In this work, we construct a monacolin J producing strain via RNA interference to achieve one-step fermentation to obtain simvastatin. The lovF gene silencing plasmid pMHJ137 was constructed and transformed into Aspergillus terreus by Agrobacterium tumefaciens mediated transformation method. Precursor DMB-S-MMP was supplied during the fermentation to screen positive strains of transformants; which also further confirmed the simvastatin producing capability of MJ1-24 by one-step fermentation. Strain MJ1-24 produced monacolin J rather than lovastatin, and the feeding of DMB-S-MMP resulted in the generation of simvastatin. This study suggested that RNAi can efficiently silence the lovF gene of A. terreus and promote the simvastatin production by one-step fermentation.
3-Mercaptopropionic Acid
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analogs & derivatives
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Aspergillus
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chemistry
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genetics
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Fermentation
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Industrial Microbiology
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Naphthalenes
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chemistry
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RNA Interference
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Simvastatin
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chemistry
2.Biosynthesis of poly (3-mercaptopropionate) and poly (3-mercaptopropionate-co-3-hydroxybutyrate) with recombinant Escherichia coli.
Shuang-Jiang LIU ; Tina LÜTKE-EVERSLOH ; Alexander STEINBÜCHEL
Chinese Journal of Biotechnology 2003;19(2):195-199
Polythioesters newly emerged as a type of novel polymer and they have showed great potential for application in industries. In this study, genes of butyrate kinase (buk) and phosphotransbutyrylase (ptb) from Clostridium acetobutylicum, and poly (3-hydroxybutyrate) (PHB) synthase gene from Thiocapsa pfennigii were used for construction of a metabolic pathway to synthesize the polythioesters. When 3-mercaptopropionate and 3-hydroxybutyrate were fed, poly (3-mercaptopropoinate) [poly (3MP)] and poly(3-mercaptopropionate-co-3-hydroxybutyrate) [poly(3MP-co-3HB)] were synthesized by recombinant Escherichia coli JM109 (pBPP1) harboring the constructed metabolic pathway. Results indicated clearly that all these genes are necessary for the synthesis of poly(3MP) and poly(3MP-co-3HB).
3-Hydroxybutyric Acid
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chemistry
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3-Mercaptopropionic Acid
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chemistry
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Acyltransferases
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genetics
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metabolism
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Escherichia coli
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genetics
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metabolism
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Gas Chromatography-Mass Spectrometry
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Models, Biological
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Molecular Weight
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Phosphate Acetyltransferase
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genetics
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metabolism
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Phosphotransferases (Carboxyl Group Acceptor)
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genetics
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metabolism
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Plasmids
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Polymers
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chemistry
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metabolism
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Spectrophotometry, Infrared