废弃毕赤酵母核糖核酸提取条件的优化
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摘要
为了确定废弃毕赤酵母提取核糖核酸的提取工艺条件,首先通过单因素试验研究了溶液pH、酵母浓度、抽提温度和抽提时间对毕赤酵母核糖核酸提取的影响,然后采用正交试验分析法对提取核糖核酸的影响因素进行了优化。优化结果表明,毕赤酵母核糖核酸提取的最佳工艺条件为:酵母浓度为3%,pH=10,抽提温度为100℃,抽提时间为2.5 h,在此条件下,核糖核酸的提取率可达3.11%。
To determine the extraction conditions of RNA from waste Pichia pastoris, the effects of yeast concentration, reaction time, reaction temperature and solution pH on the extraction rate of RNA from P. pastoris were firstly studied by single-factor-test. Then the extraction condition was further optimized by orthogonal experimental design. The results showed the optimum extraction conditions of RNA from P. pastoris was yeast concentration of 3%, solution pH 10, reaction temperature 100 ℃ and extraction time 2.5 h.The RNA extraction rate could be 3.11% at the optimized conditions.
To determine the extraction conditions of RNA from waste Pichia pastoris, the effects of yeast concentration, reaction time, reaction temperature and solution pH on the extraction rate of RNA from P. pastoris were firstly studied by single-factor-test. Then the extraction condition was further optimized by orthogonal experimental design. The results showed the optimum extraction conditions of RNA from P. pastoris was yeast concentration of 3%, solution pH 10, reaction temperature 100 ℃ and extraction time 2.5 h.The RNA extraction rate could be 3.11% at the optimized conditions.
引文
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[3]倪晓丰,赵宾,王东旭,等.硫酸二乙酯化学诱变选育高核糖核酸酿酒酵母及培养基组成优化[J].中国酿造,2018,37(8):32-36.
[4]蒋雪薇,岳希洁,陈代文,等.产朊假丝酵母核酸高产株的选育及其发酵性能研究[J].食品科学,2017,38(10):149-154.
[5]朱俊东,黄国荣,糜漫天,等.啤酒酵母中提取核糖核酸的研究[J].食品科学,2006,27(2):160-162.
[6]胡刚,孙军勇,蔡国林,等.浓盐法提取啤酒废酵母核糖核酸的研究[J].中国酿造,2009,28(7):112-114.
[7] Li C, Lin Y, Zheng X, et al. Combined strategies for improving expression of Citrobacter amalonaticus phytase in Pichia pastoris[J]. BMC Biotechnology, 2015, 15:88.
[8] Hmida-Sayari A, Elgharbi F, Farhat A, et al. Overexpression and biochemical characterization of a thermostable phytase from Bacillus subtilis US417 in Pichia pastoris[J].Molecular Biotechnology, 2014, 56(9):839-848.
[9]潘阳,吴丹,吴敬.利用游离型表达质粒强化毕赤酵母表达木聚糖酶[J].生物工程学报,2018, 34(5):712-721.
[10] Zhu W, Gong G, Pan J, et al. High level expression and purification of recombinant human serum albumin in Pichia pastoris[J]. Protein Expression and Purification, 2018,147:61-68.
[11]朱红蕾,杨海龙.从废弃毕赤酵母提取甘露聚糖的研究[J].温州大学学报(自然科学版),2012,33(1):17-21.
[12]徐慧,刘建军,赵祥颖,等.弱碱盐法提取核糖核酸工艺的研究[J].酿酒,2008,35(3):60-62.
[13]钟平,张熊禄,邱承洲.稀碱裂解法从啤酒糟中提取核糖核酸的研究[J].化学世界,2004,(3):138-140.