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Background: Isatropolone A and C, produced by Streptomyces sp. CPCC 204095, belong to an unusual class of non-benzenoid aromatic compounds and contain a rare seven-membered ring structure. Isatropolone A exhibits potent activity against Leishmania donovani, comparable to the only oral drug miltefosine. However, its variably low productivity represents a limitation for this lead compound in the future development of new anti-leishmaniasis drugs to meet unmet clinical needs. Results: Here we first elucidated the regulatory cascade of biosynthesis of isatropolones, which consists of two SARP family regulators, IsaF and IsaJ. Through a series of in vivo and in vitro experiments, IsaF was identified as a pathway-specific activator that orchestrates the transcription of the gene cluster essential for isatropolone biosynthesis. Interestingly, IsaJ was found to only upregulate the expression of the cytochrome P450 monooxygenase IsaS, which is crucial for the yield and proportion of isatropolone A and C. Through targeted gene deletions of isaJ or isaS, we effectively impeded the conversion of isatropolone A to C. Concurrently, the facilitation of isaF overexpression governed by selected promoters, prompted the comprehensive activation of the production of isatropolone A. Furthermore, meticulous optimization of the fermentation parameters was conducted. These strategies culminated in the attainment of an unprecedented maximum yield—980.8 mg/L of isatropolone A—achieved in small-scale solid-state fermentation utilizing the genetically modified strains, thereby establishing the highest reported titer to date. Conclusion: In Streptomyces sp. CPCC 204095, the production of isatropolone A and C is modulated by the SARP regulators IsaF and IsaJ. IsaF serves as a master pathway-specific regulator for the production of isatropolones. IsaJ, on the other hand, only dictates the transcription of IsaS, the enzyme responsible for the conversion of isatropolone A and C. By engineering the expression of these pivotal genes, we have devised a strategy for genetic modification aimed at the selective and high-yield biosynthesis of isatropolone A. This study not only unveils the unique regulatory mechanisms governing isatropolone biosynthesis for the first time, but also establishes an essential engineering framework for the targeted high-level production of isatropolone A. [ABSTRACT FROM AUTHOR]

Simple Summary: Research on antioxidant status and lipid metabolism in pigeon squabs as meat type poultry is limited. The objective of this study was to explore the age-related changes in the antioxidant status and lipid metabolism of pigeon squabs (Columba livia). Ten squabs were randomly selected, out of 400 hatched squabs, on the following days: day of hatching (DOH) and days 7 (D7), 14 (D14), 21 (D21) post-hatch, respectively. BW, serum lipid levels, antioxidant capacity indices, lipid metabolism-related enzyme activities, lipid metabolism-related gene expression, and liver morphology were determined. Results indicated that the phase from DOH to D14 was a growth spurt, especially in the first seven days. The antioxidant capacity of squabs had a continuous decline from DOH to D14. Besides, changes regarding lipid metabolism mainly occurred in the phase from DOH to D14. The first seven days mainly showed less lipid breakdown, while the second seven days displayed more complicated lipid metabolism. The results obtained from this study suggested that, in the pigeon industry, it is better to take nutritional manipulation in squabs before D14. The objective of this study was to evaluate the age-related changes in antioxidant status and the lipid metabolism of pigeon squabs (Columba livia), by determining the BW, antioxidant indices, serum lipid levels, lipid metabolism-related enzyme activities, lipid metabolism-related gene expression, and liver morphology in squabs. Ten squabs were randomly selected and sampled on the day of hatching (DOH), days 7 (D7), 14 (D14) and 21 (D21) post-hatch, respectively. The results showed that BW of squabs increased linearly from DOH to D21. The minimum fold of BW gain was observed in the phase from D14 to D21. Serum triglyceride and free fatty acid levels displayed linear and quadratic trends as age increased, with these maximum responses in D14. Serum low-density lipoprotein cholesterol level responded to age linearly and quadratically with the minimum in D14. Serum high-density lipoprotein cholesterol level and the ratio of high-density lipoprotein cholesterol to low-density lipoprotein cholesterol increased linearly with age, whereas the very low-density lipoprotein cholesterol level decreased linearly. The activities of glutathione peroxidase, catalase, and superoxide dismutase in liver displayed linear and quadratic trends as age increased, with these minimum responses in D14. Hepatic malondialdehyde concentration responded to age linearly and quadratically, with the maximum in D14. Activities of lipoprotein lipase, hepatic lipase, and 3-hydroxy-3-methyl glutaryl coenzyme A reductase in liver responded to age linearly and quadratically, with these minimum responses in D14. Hepatic hormone-sensitive lipase activity displayed linear and quadratic trends as age increased with the maximum in D14. Hepatic acetyl CoA carboxylase activity on D14 was significantly lower than squabs on DOH and D7. Hepatic carnitine palmitoyltransferase 1 mRNA expression responded to age linearly and quadratically, with minimum response in D14. Hepatic mRNA expression of acetyl CoA carboxylase and fatty acid synthetase increased linearly with age. Hepatic Oil-Red-O staining area displayed a quadratic trend as age increased, with the maximum response in D14. In conclusion, the phase from DOH to D14 was a crucial development stage for growth, antioxidant status and lipid metabolism in pigeon squabs. The results suggest it is better to take nutritional manipulation in squabs before D14. [ABSTRACT FROM AUTHOR]

Simple Summary: In the current study, we hypothesize that the In ovo concurrent injection of methionine (Met) and cysteine (Cys) may have positive effects on embryonic development, insulin-like growth factor-I (IGF-I) and toll-like receptor-4 (TLR4) gene expression, antioxidant status, biochemical profile, and jejunum histomorphometry of newly hatched broiler chicks exposed to heat stress during the incubation period. As indicated, the In ovo injection of Met and Cys resulted in increased hatch weight, higher levels of total antioxidant capacity, and glutathione activity in the serum and tissues. At the same time, a decrease in the level of heat shock protein-90 and an increase in TLR4 and IGF-I expression in the tissues was observed. In addition, the In ovo injection of Met and Cys improved jejunum histomorphometric parameters of newly hatched broiler chicks. Generally, the concurrent In ovo injection of Met and Cys improved broiler embryonic development, antioxidant status, TLR4, and IGF-I expression and jejunum histomorphometric parameters in newly hatched broiler chicks exposed to heat stress conditions during incubation. Sulfur amino acids are typically the first-limiting amino acids (AA) used in protein metabolism in poultry. Therefore, we hypothesized that their utilization in the pre-hatch period would affect embryonic development, IGF-I and TLR4 gene expression, antioxidant status, serum biochemical profile, and jejunum histomorphometry of newly hatched Ross broiler chicks incubated under heat stress conditions. A total of 150 fertile broiler eggs were subjected to heat stress (39.6 °C for 6 h/d) from d10 until d18 and injected at d 17.5 of incubation with methionine and cysteine (Met-Cys) at a dose of 5.90 mg l-methionine plus 3.40 mg l-cysteine. The effects of Met-Cys administration were examined and compared with the control (Non-injected group) and 0.75% NaCl injected group. The results showed that no significant differences among all groups in serum protein profiles (total protein, albumin, globulin, and albumin/globulin ratio) and creatine kinase were observed. The level of heat shock protein-90 was decreased with Met-Cys In ovo injection. The In ovo injection of Met-Cys also improved the values of total antioxidants capacity and glutathione in examined tissues. At the same time, an increase in fold change mRNA abundance of IGF-I and TLR4 was observed after Met-Cys injection in tested tissues. Finally, an increase of 29% in villus area was found after Met-Cys injection compared to the control group. In conclusion, the In ovo injection of Met-Cys resulted in improved embryonic development, IGF-I and TLR4 gene expression, antioxidant status and jejunum histomorphometry of newly hatched broiler chicks exposed to heat stress during incubation. [ABSTRACT FROM AUTHOR]