APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2023
·Vol. 107
·期 11
·3745-3761
·
IF 5
Xu Chenglong,Lin Wenxi,Chen Yunni,Gao Boliang,Zhang Zhibin,Zhu Du
Perylenequinones (PQs) are a class of natural polyketides used as photodynamic therapeutics. Heat stress (HS) is an important environmental factor affecting secondary metabolism of fungi. This study investigated the effects of HS treatment on PQs biosynthesis of Shiraia sp. Slf14(w) and the underlying molecular mechanism. After the optimization of HS treatment conditions, the total PQs amount reached 577 ± 34.56 mg/L, which was 20.89-fold improvement over the control. Also, HS treatment stimulated the formation of intracellular nitric oxide (NO). Genome-wide analysis of Shiraia sp. Slf14(w) revealed iNOSL and cNOSL encoding inducible and constitutive NOS-like proteins (iNOSL and cNOSL), respectively. Cloned iNOSL in Escherichia coli BL21 showed higher nitric oxide synthase (NOS) activity than cNOSL, and the expression level of iNOSL under HS treatment was observably higher than that of cNOSL , suggesting that iNOSL is more responsible for NO production in the HS-treated strain Slf14(w) and may play an important role in regulating PQs biosynthesis. Moreover, the putative biosynthetic gene clusters for PQs and genes encoding iNOSL and nitrate reductase (NR) in the HS-treated strain Slf14(w) were obviously upregulated. PQs biosynthesis and efflux stimulated by HS treatment were significantly inhibited upon the addition of NO scavenger, NOS inhibitor, and NR inhibitor, indicating that HS-induced NO, as a signaling molecule, triggered promoted PQs biosynthesis and efflux. Our results provide an effective strategy for PQs production and contribute to the understanding of heat shock signal transduction studies of other fungi.
Key points
• PQs titer of Shiraia sp. Slf14(w) was significantly enhanced by HS treatment . • HS-induced NO was first reported to participate in PQs biosynthetic regulation . • Novel inducible and constitutive NOS-like proteins (iNOSL and cNOSL) were obtained and their NOS activities were determined .
研究领域:
微生物学
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分子生物学
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药理学
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
·
2025
·Vol. 41
·期 10
·1-12
·
IF 4.2
Sun Ying,Hua Qijing,Cai Yinglin,Dong Yamin,Xiao Yiwen,Bao Jianyin,Zhu Du,Liu Bin,Gao Boliang
Metal ions critically regulate microbial secondary metabolism. This study investigated the regulatory roles of metal ions in Shiraia sp. Slf14, an endophytic fungus with exceptional perylenequinone (PQ) biosynthesis capacity. Among eight ions tested, Na⁺, K⁺, Mg²⁺, and high Ca²⁺ (1.0 g/L) markedly enhanced PQ yields, while Mn²⁺, Cu²⁺, Zn²⁺, and Fe³⁺ (≥ 0.5 g/L) exerted strong inhibition. Na⁺ emerged as the optimal elicitor, boosting production to 504.82 ± 8.62 mg/L (+ 57.96%) at 1.0 g/L without cytotoxicity. Na⁺ exhibited concentration- and time-dependent biphasic regulation. Critical innovation was achieved at 4 g/L Na⁺ added on day 4 (early-log phase), maximizing total PQ yield (1836.95 ± 45.21 mg/L, 5.75 × control), while concentrations > 12 g/L or late addition (≥ day 8) suppressed biosynthesis. Component-specific sensitivities were observed among five PQs. Furthermore, Na⁺ activated the calcineurin pathway: stress-induced Cam/Cna triggered calcineurin-mediated Crz1 nuclear translocation, upregulating key PQ genes ( pksI , omef , hydroxylase ). Concurrent induction of ena1 (Na⁺ exporter) and Ca²⁺-ATPase maintained ion homeostasis. In conclusion, our work pointed out monovalent cations (Na⁺/K⁺) as potent PQ biosynthetic stimulators in filamentous fungi and establish the Na⁺-calcineurin-PQ axis, offering a scalable strategy to overcome production bottlenecks in pharmaceutical PQ manufacturing.
研究领域:
微生物学
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真菌生物学
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次级代谢产物
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信号转导
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药物生物技术
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分子生理学
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天然产物化学
✓ 已验证
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2022
·Vol. 106
·期 7
·2619-2636
·
IF 5.56
Chen Yunni,Xu Chenglong,Yang Huilin,Liu Zhenying,Zhang Zhibin,Yan Riming,Zhu Du
Abstract Perylenequinones (PQ) are natural polyketides used as anti-microbial, -cancers, and -viral photodynamic therapy agents. Herein, the effects of L-arginine (Arg) on PQ biosynthesis of Shiraia sp. Slf14(w) and the underlying molecular mechanism were investigated. The total content of PQ reached 817.64 ± 72.53 mg/L under optimal conditions of Arg addition, indicating a 30.52-fold improvement over controls. Comparative transcriptome analysis demonstrated that Arg supplement promoted PQ precursors biosynthesis of Slf14(w) by upregulating the expression of critical genes associated with the glycolysis pathway, and acetyl-CoA and malonyl-CoA synthesis. By downregulating the expression of genes related to the glyoxylate cycle pathway and succinate dehydrogenase, more acetyl-CoA flow into the formation of PQ. Arg supplement upregulated the putative biosynthetic gene clusters for PQ and activated the transporter proteins (MFS and ABC) for exudation of PQ. Further studies showed that Arg increased the gene transcription levels of nitric oxide synthase (NOS) and nitrate reductase (NR), and activated NOS and NR, thus promoting the formation of nitric oxide (NO). A supplement of NO donor sodium nitroprusside (SNP) also confirmed that NO triggered promoted biosynthesis and efflux of PQ. PQ production stimulated by Arg or/and SNP can be significantly inhibited upon the addition of NO scavenger carboxy-PTIO, NOS inhibitor N ω -nitro-L-arginine, or soluble guanylate cyclase inhibitor NS-2028. These results showed that Arg-derived NO, as a signaling molecule, is involved in the biosynthesis and regulation of PQ in Slf14(W) through the NO-cGMP-PKG signaling pathway. Our results provide a valuable strategy for large-scale PQ production and contribute to further understanding of NO signaling in the fungal metabolite biosynthesis. Key points • PQ production of Shiraia sp. Slf14(w) was significantly improved by L-arginine addition. • Arginine-derived NO was firstly reported to be involved in the biosynthesis and regulation of PQ. • The NO-cGMP-PKG signaling pathway was proposed for the first time to participate in PQ biosynthesis.
研究领域:
微生物学
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生物化学
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药理学
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分子生物学
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真菌代谢
✓ 已验证
DOI
PubMed
高影响因子
3 Biotech
·
2020
·Vol. 10
·期 5
·1-13
·
IF 1.798
Liu Zhengying,Bao Jianying,Yang Huilin,Zhang Zhibin,Yan Riming,Zhu Du
Perylenequinones (PQ), a class of naturally occurring polypeptides, are widely used as a clinical drug for treating skin diseases and as a photodynamic therapy against cancers and viruses. In this study, the effects of different carbon sources on PQ biosynthesis by Shiraia sp. Slf14 were compared, and the underlying molecular mechanism of fructose as the sole carbon to enhance PQ production was investigated by transcriptome analysis. The results indicated that fructose enhanced PQ yield to 1753.64 mg/L, which was 1.73-fold higher than that obtained with glucose. Comparative transcriptome analysis demonstrated that most of the upregulated genes were related to transport systems, energy and central carbon metabolism in Shiraia sp. Slf14 cultured in fructose. The genes involved in glycolysis and pentose phosphate pathways, and encoding citrate synthase, ATP-citrate lyase, and acetyl-CoA carboxylase were substantially upregulated, resulting in increased overall acetyl-CoA and malonyl-CoA production. However, genes involved in gluconeogenesis, glyoxylate cycle pathway, and fatty acid synthesis were significantly downregulated, resulting in higher acetyl-CoA influx for PQ formation. In particular, the putative PQ biosynthetic cluster was upregulated in Shiraia sp. Slf14 cultured in fructose, leading to a significant increase in PQ production. The results of real-time qRT-PCR and related enzyme activities were also consistent with those of transcriptome analysis. These findings provide a remarkable insight into the underlying mechanism of PQ biosynthesis and pave the way for improvements in PQ production by Shiraia sp. Slf14.
研究领域:
分子生物学
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微生物学
·
生物技术
·
药学
✓ 已验证
DOI
PubMed
高影响因子
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