Abstract
This study characterized the biosynthetic pathway of the secondary metabolite 1-deoxynojirimycin (DNJ) from Streptomyces lavendulae. The results revealed that glucose was a preferable precursor for DNJ synthesis, and its carbon skeleton underwent a C2-N-C6 cyclization reaction during synthesis. The biosynthetic pathway was related to the glycolysis pathway, and started from fructose-6-phosphate, and involved amination, dephosphorylation, oxidation, cyclization, dehydration, and reduction reaction steps, yielding DNJ. Then, based on clarified biosynthetic pathway information, precursors, analogs, and metabolism inhibitors were used as novel regulators to enhance the production of DNJ. The results demonstrated that the titer of DNJ could reach 296.56 mg/L, which was 3.3-fold higher than that of a control group (90 mg/L) when sodium citrate (0 h, 5 g/L), sorbose (0 h, 1 g/L), iodoacetic acid (20 h, 50 mg/L), and glucose (26 h, 7 g/L) were added during the fermentation process. This study provides a new understanding of the biosynthetic pathway of DNJ, and also provides an efficient strategy to regulate the production of DNJ based on this biosynthetic pathway, which is a new perspective for the regulation of other secondary metabolites.
Highlights
1-Deoxynojirimycin (DNJ), a type of alkaloid secondary metabolite, was first isolated from mulberry trees by Yagi et al (1976)
Key genes involved in DNJ biosynthesis in mulberry (Morus alba L.) have been identified through transcriptome analysis, and the biosynthetic pathway starting from aspartate has been outlined (Wang et al, 2018)
The current study reveals novel information pertaining to the biosynthetic pathway of DNJ, and provides an efficient strategy to regulate the production of DNJ, which may be relevant for the regulation of other secondary metabolites
Summary
1-Deoxynojirimycin (DNJ), a type of alkaloid secondary metabolite, was first isolated from mulberry trees by Yagi et al (1976). Given the findings of the above studies, it can be suggested that the synthetic routes of DNJ are diverse, with differences between microorganisms and plants, and even plants themselves (M. alba L. and C. communis) Streptomyces lavendulae, another mainstream DNJ-producing strain, has been reported in many studies (Ezure et al, 1985; Kojima et al, 1995; Wei et al, 2011). There are no recent studies investigating the regulation of DNJ production using feeding experiments with specific inhibitors and precursors from biosynthetic pathways. The current study attempted to elucidate the biosynthetic pathway of DNJ in S. lavendulae through precursor feeding experiments, analyzing carbon skeleton conversion using isotope-labeled glucose, and identifying related metabolic intermediates through hydrophilic interaction chromatography with tandem mass spectrometry (HILIC-MS/MS) techniques. The current study reveals novel information pertaining to the biosynthetic pathway of DNJ, and provides an efficient strategy to regulate the production of DNJ, which may be relevant for the regulation of other secondary metabolites
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