Presentation Information
[P01-092]Biosynthetic investigation of azoxyalkene compound KA57A and maniwamycin G, using comparative genomic analysis
○Sari Sumida1, Kaito Fukumori1, Yu Tanaka1, Haruka Nagano1, Ayaka Tatsukawa1, Yojiro Anzai2, Aiko Teshima1, Kenji Arakawa1 (1. Hiroshima Univ. (Japan), 2. Toho Univ. (Japan))
Keywords:
azoxyalkene,Streptomyces,biosynthesis
[Purpose]
Streptomyces species have an ability to produce many useful secondary metabolites, such as antibiotics, and their genomes contain more than 30 genes involved in secondary metabolite biosynthesis. However, under standard culture conditions, only about 20% of these genes are expressed, finding that approximately 80% of the potential secondary metabolites remain unexpressed. To identify potential secondary metabolites, we constructed the triple mutant strain KA57 of Streptomyces rochei 7434AN4. This mutant accumulated azoxyalkene compound, KA57A1, 2). Additionally, maniwamycin G 3), a structural analog of this compound, was isolated from a different strain, TOHO-M025. These compounds exhibit quorum sensing inhibitory activity against Chromobacterium violaceum. Compounds containing azoxy bonds, such as these, are rare framework, and many possess unique biological activities; however, their whole biosynthetic pathways remain largely unexplored. Therefore, our objective was to elucidate the complete biosynthetic pathways of the two azoxyalkene compounds, KA57A and maniwamycin G, and to deepen our understanding of their structural diversity.
[Method and Results]
First, we predicted the biosynthetic pathways of KA57A and maniwamycin G through comparative genomic analysis. Next, we constructed several knockout strains of genes possibly involved in biosynthesis and performed metabolite analysis. Among them, a notable accumulation of UV active compounds was observed in a mutant of SRO_1821 (ΔvlmJ (kinase) homolog). This mutant TK01 accumulated novel hydrazide-alkene compound KA57D3. Furthermore, a trace amount of KA57A (below 10% yield of the parent strain KA57) was detected, indicating the other kinase homolog(s) is(are) responsible for phosphorylation to convert KA57A. Regarding TOHO-M025, we isolated two-UV-inactive compounds, which were determined to be γ-butyrolactone molecules, which may act as signaling molecules in this strain. Details will be described in this paper.
[Reference]
1) Kunitake H, Hiramatsu T, Kinashi H, Arakawa K.
“Isolation and biosynthesis of an azoxyalkene compound produced by a multiple gene disruptant of Streptomyces rochei”
ChemBioChem,16, 2237-2243 (2015).
2) Tanaka Y, Nagano H, Okano M, Kishimoto T, Tatsukawa A, Kunitake H, Fukumoto A, Anzai Y, and Arakawa K.
“Isolation of hydrazide-alkenes with different amino-acid origins from an azoxy-alkene-producing mutant of Streptomyces rochei 7434AN4”
Journal of Natural Products, 86, 2185-2192 (2023).
3) Tatsukawa A, Tanaka Y, Nagano H, Fukumoto A, Anzai Y, and Arakawa K.
“Isolation, biosynthetic investigation, and biological evaluation of maniwamycin G, an azoxyalkene compound from Streptomyces sp. TOHO-M025”
Journal of Natural Products, 85, 1867-1871 (2022).
Streptomyces species have an ability to produce many useful secondary metabolites, such as antibiotics, and their genomes contain more than 30 genes involved in secondary metabolite biosynthesis. However, under standard culture conditions, only about 20% of these genes are expressed, finding that approximately 80% of the potential secondary metabolites remain unexpressed. To identify potential secondary metabolites, we constructed the triple mutant strain KA57 of Streptomyces rochei 7434AN4. This mutant accumulated azoxyalkene compound, KA57A1, 2). Additionally, maniwamycin G 3), a structural analog of this compound, was isolated from a different strain, TOHO-M025. These compounds exhibit quorum sensing inhibitory activity against Chromobacterium violaceum. Compounds containing azoxy bonds, such as these, are rare framework, and many possess unique biological activities; however, their whole biosynthetic pathways remain largely unexplored. Therefore, our objective was to elucidate the complete biosynthetic pathways of the two azoxyalkene compounds, KA57A and maniwamycin G, and to deepen our understanding of their structural diversity.
[Method and Results]
First, we predicted the biosynthetic pathways of KA57A and maniwamycin G through comparative genomic analysis. Next, we constructed several knockout strains of genes possibly involved in biosynthesis and performed metabolite analysis. Among them, a notable accumulation of UV active compounds was observed in a mutant of SRO_1821 (ΔvlmJ (kinase) homolog). This mutant TK01 accumulated novel hydrazide-alkene compound KA57D3. Furthermore, a trace amount of KA57A (below 10% yield of the parent strain KA57) was detected, indicating the other kinase homolog(s) is(are) responsible for phosphorylation to convert KA57A. Regarding TOHO-M025, we isolated two-UV-inactive compounds, which were determined to be γ-butyrolactone molecules, which may act as signaling molecules in this strain. Details will be described in this paper.
[Reference]
1) Kunitake H, Hiramatsu T, Kinashi H, Arakawa K.
“Isolation and biosynthesis of an azoxyalkene compound produced by a multiple gene disruptant of Streptomyces rochei”
ChemBioChem,16, 2237-2243 (2015).
2) Tanaka Y, Nagano H, Okano M, Kishimoto T, Tatsukawa A, Kunitake H, Fukumoto A, Anzai Y, and Arakawa K.
“Isolation of hydrazide-alkenes with different amino-acid origins from an azoxy-alkene-producing mutant of Streptomyces rochei 7434AN4”
Journal of Natural Products, 86, 2185-2192 (2023).
3) Tatsukawa A, Tanaka Y, Nagano H, Fukumoto A, Anzai Y, and Arakawa K.
“Isolation, biosynthetic investigation, and biological evaluation of maniwamycin G, an azoxyalkene compound from Streptomyces sp. TOHO-M025”
Journal of Natural Products, 85, 1867-1871 (2022).
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