Impact on strain growth and butenyl-spinosyn biosynthesis by overexpression of polynucleotide phosphorylase gene in Saccharopolyspora pogona

Polynucleotide phosphorylase is a highly conserved protein found in bacteria and fungi that can regulate the transcription of related enzymes involved in amino acid metabolism, organic acid metabolism, and cell biosynthesis. We studied the effect of polynucleotide phosphorylase on Saccharopolyspora...

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Published in:Applied microbiology and biotechnology 2018-09, Vol.102 (18), p.8011-8021
Main Authors: Li, Li, Rang, Jie, He, Haochen, He, Siying, Liu, Zhudong, Tang, Jianli, Xiao, Jie, He, Lian, Hu, Shengbiao, Yu, Ziquan, Ding, Xuezhi, Xia, Liqiu
Format: Article
Language:English
Subjects:
Actinomycetes
Amino acids
Applied Microbial and Cell Physiology
Biomedical and Life Sciences
Biosynthesis
Biotechnology
Fungi
Gene expression
Genetic aspects
High-performance liquid chromatography
Life Sciences
Liquid chromatography
Metabolism
Metabolites
Microbial Genetics and Genomics
Microbiology
Observations
Organic acids
Phosphorylase
Polynucleotide phosphorylase
Proteins
Recombination
Saccharopolyspora
Secondary metabolites
Spores
Strains (organisms)
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Summary:Polynucleotide phosphorylase is a highly conserved protein found in bacteria and fungi that can regulate the transcription of related enzymes involved in amino acid metabolism, organic acid metabolism, and cell biosynthesis. We studied the effect of polynucleotide phosphorylase on Saccharopolyspora pogona ( S. pogona ) growth and the synthesis of secondary metabolites. First, we generated the overexpression vector pOJ260- P ermE -pnp via overlap extension PCR. The vector pOJ260- P ermE -pnp was then introduced into S. pogona by conjugal transfer, thereby generating the recombination strain S. pogona -Pnp. Results showed that engineering strains possessed higher biomass than those of the wild-type strains. Moreover, the ability of these strains to produce spores on solid medium was stronger than that of the wild-type strains. HPLC results revealed that the butenyl-spinosyn yield in S. pogona -Pnp increased by 1.92-fold compared with that of S. pogona alone. These findings revealed that overexpression of polynucleotide phosphorylase effectively promoted butenyl-spinosyn biosynthesis in S. pogona . This result may be extended to other Streptomyces for strain improvement.
ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-018-9178-z