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表达豆血红蛋白的酿酒酵母产血红素条件优化
Optimization of heme formation conditions of leghemoglobin in Saccharomyces cerevisiae
【摘要】 为提升豆血红蛋白在酿酒酵母中生产血红素的发酵产率,以发酵液中血红素浓度为指标,在单因素实验基础上,采用响应曲面法,以5-氨基乙酰丙酸添加量、发酵时间和发酵温度为影响因素,以发酵液中血红素浓度为响应值建立二次回归方程,通过响应面分析得到各因素的优化组合。研究结果表明:5-氨基乙酰丙酸添加量对血红素浓度的影响最大,其次是发酵温度,最后为发酵时间。综合考虑时间、能耗及血红素产率等因素,当发酵培养基的pH为5,向发酵培养基中添加硫酸亚铁和5-氨基乙酰丙酸至终浓度分别为10μmol/L和1.35 mmol/L,诱导剂半乳糖质量浓度为80 g/L,发酵温度为28℃,发酵时间为50 h时发酵效果较好。在此优化条件下,血红素浓度提升至9.036μmol/L,获得了较高发酵产率,未来可以进一步优化,将该法推广至产业应用。
【Abstract】 To improve the production of heme by leghemoglobin in Saccharomyces cerevisiae, the concentration of heme in fermentation liquid was taken as the index, and the response surface method was adopted on the basis of a single factor test, with the addition of 5-aminolevulinic acid, fermentation time, and fermentation temperature as the influencing factors. The quadratic regression equation was established with the concentration of heme in fermentation solution as the response value, and the optimal combination was obtained by response surface analysis. The results showed that the content of 5-aminolevulinic acid had the greatest effect on the heme concentration, followed by fermentation temperature, and finally fermentation time. Considering factors such as time, energy consumption, and yield: when the pH value of fermentation medium was 5, ferric sulfate and 5-aminolevulinic acid were added to the fermentation medium to the final concentration of 10 μmol/L and 1.35 mmol/L, respectively, the concentration of inducer galactose was 80 g/L, the fermentation temperature was 28 ℃, and the fermentation time was 50 h, the fermentation effect was better. Under this optimized condition, the heme concentration was increased to 9.036 μmol/L, and a high yield was obtained. Further optimization could lead to a promising industrialization scenario.
- 【文献出处】 发酵科技通讯 ,Bulletin of Fermentation Science and Technology , 编辑部邮箱 ,2024年04期
- 【分类号】TQ920.6
- 【下载频次】21