دوره 36، شماره 2 - ( 3-1404 )
جلد 36 شماره 2 صفحات 122-114 |
برگشت به فهرست نسخه ها
چکیده: (594 مشاهده)
Background Methionine, an essential sulfur-containing amino acid, plays a critical role in detoxification via methylation and must be obtained through the diet, as humans cannot synthesize it. This study aimed to identify the met gene and analyze the kinetics of methionine synthesis in soil-derived thermophilic bacteria.
Methods Two hundred soil samples were collected from tree-adjacent areas along Khordin Boulevard, Tehran. After heat treatment and culturing, isolates underwent microscopic, biochemical, and molecular characterization. DNA was extracted using a specialized kit, and the met gene was identified via multiplex PCR and gel electrophoresis. Gene expression was quantified using quantitative real-time PCR (qRT-PCR) under three temperatures (25°C, 35°C, 45°C) with ammonium nitrate (nitrogen source) and glucose (carbon source). Data were analyzed by one-way ANOVA using SPSS v.22 and reported as mean ± SD at a significance level of p < 0.05.
Results Bacilli accounted for 30% of isolates from Tehran’s urban soil. The met gene was detected in only 6.66% of bacilli. Significant differences in met expression were observed between treated and control groups across all temperatures and nutrient conditions (p < 0.05). The most excellent suppression of met expression occurred at 45°C (2.342-fold decrease), while 25°C showed the least reduction (1.649-fold). Glucose and ammonium nitrate synergistically reduced expression (1.914- and 1.834-fold, respectively).
Conclusion Temperature and carbon/nitrogen sources modulate met gene expression, thereby influencing methionine synthesis in soil bacilli. Optimal suppression occurred at 45°C with a combination of glucose/ammonium nitrate, suggesting environmental regulation of this metabolic pathway. In contrast, the lowest suppression of met expression was observed at 25°C.
Methods Two hundred soil samples were collected from tree-adjacent areas along Khordin Boulevard, Tehran. After heat treatment and culturing, isolates underwent microscopic, biochemical, and molecular characterization. DNA was extracted using a specialized kit, and the met gene was identified via multiplex PCR and gel electrophoresis. Gene expression was quantified using quantitative real-time PCR (qRT-PCR) under three temperatures (25°C, 35°C, 45°C) with ammonium nitrate (nitrogen source) and glucose (carbon source). Data were analyzed by one-way ANOVA using SPSS v.22 and reported as mean ± SD at a significance level of p < 0.05.
Results Bacilli accounted for 30% of isolates from Tehran’s urban soil. The met gene was detected in only 6.66% of bacilli. Significant differences in met expression were observed between treated and control groups across all temperatures and nutrient conditions (p < 0.05). The most excellent suppression of met expression occurred at 45°C (2.342-fold decrease), while 25°C showed the least reduction (1.649-fold). Glucose and ammonium nitrate synergistically reduced expression (1.914- and 1.834-fold, respectively).
Conclusion Temperature and carbon/nitrogen sources modulate met gene expression, thereby influencing methionine synthesis in soil bacilli. Optimal suppression occurred at 45°C with a combination of glucose/ammonium nitrate, suggesting environmental regulation of this metabolic pathway. In contrast, the lowest suppression of met expression was observed at 25°C.
نوع مطالعه: پژوهشي(توصیفی- تحلیلی) |
موضوع مقاله:
میکروبیولوژی
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