转基因大豆水酶法制油过程中内、外源基因的分布研究
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摘要
以挤压膨化过的转基因大豆为原料,大豆油的提取率为指标,在单因素试验的基础上,使用微滴式数字PCR仪对水酶法提取转基因大豆油的工艺过程中内、外源基因的分布进行研究,探索转基因大豆内、外源基因的降解规律。通过单因素试验确定水酶法提取转基因大豆油的最佳工艺条件为:料液比1∶6(g/mL),碱性蛋白酶加酶量1.8%,酶解温度35℃,酶解时间4.0 h,酶解pH 9.0,在此条件下,通过水酶法得到的水相中转基因大豆的内源基因占65.0652%,外源基因占81.742 1%;固相中内源基因占16.892 5%,外源基因占11.284 2%;油相中内源基因占0.000 2%,外源基因占0.000 4%。
Based on the single-factor experiment,used the extruded genetically modified soybeans as raw material and the extraction rate of soybean oil as the index,used the quantitative research methods of digital PCR to study the effects to endogenous and exogenous gene in the process of oil extraction by aqueous enzymatic,furthermore,the degradation of endogenous and exogenous genes in transgenic soybeans was explored.The optimal conditions were determined as follows:the ratio of solid to liquid was 1 ∶ 6(g/mL),the amount of alkaline protease was 1.8 %,the temperature was 35 ℃,the time was 4.0 h,and the pH was 9.0.Under these conditions,the endogenous genes of transgenic soybean in the aqueous phase obtained by the aqueous enzymatic method were accounted for 65.065 2 %,the exogenous genes were accounted for 81.742 1 %;the endogenous genes were accounted for 16.892 5 % in the solid phase,and the exogenous genes were accounted for 11.284 2 %;the endogenous genes were accounted for 0.000 2 % in the oil phase,and the exogenous genes were accounted for 0.000 4 %.
Based on the single-factor experiment,used the extruded genetically modified soybeans as raw material and the extraction rate of soybean oil as the index,used the quantitative research methods of digital PCR to study the effects to endogenous and exogenous gene in the process of oil extraction by aqueous enzymatic,furthermore,the degradation of endogenous and exogenous genes in transgenic soybeans was explored.The optimal conditions were determined as follows:the ratio of solid to liquid was 1 ∶ 6(g/mL),the amount of alkaline protease was 1.8 %,the temperature was 35 ℃,the time was 4.0 h,and the pH was 9.0.Under these conditions,the endogenous genes of transgenic soybean in the aqueous phase obtained by the aqueous enzymatic method were accounted for 65.065 2 %,the exogenous genes were accounted for 81.742 1 %;the endogenous genes were accounted for 16.892 5 % in the solid phase,and the exogenous genes were accounted for 11.284 2 %;the endogenous genes were accounted for 0.000 2 % in the oil phase,and the exogenous genes were accounted for 0.000 4 %.
引文
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[14]王心刚,江连洲,李杨,等.真空挤压膨化水酶法提取大豆油的工艺研究[J].中国粮油学报,2013,28(11):28-31
[15]王卫国,朱占齐,吴兴泉,等.微波、超高压处理对转基因大豆外源基因降解的影响[J].中国粮油学报,2015,30(2):20-25
[16]吴洪洪.转基因Roundup Ready大豆外源CP4-EPSPS蛋白及内外源基因在食品加工过程中的降解变化规律[D].南京:南京农业大学,2011
[17]Guan Q,Wang X,Teng D,et al.DNA degradation of genetically modified cottonseed meal during feed processing[J].Applied Biochemistry&Biotechnology,2013,169(2):368-379
[18]王林,韩飞,李爱科,等.不同加工工艺对大豆转基因成分及调控元件的影响[J].大豆科学,2011,30(1):136-140
[19]夏义苗.一种用于提取大豆种子基因组DNA的细胞裂解液及其提取方法:201710888077.0[P].2018-1-12
[2]杨爱萍,蒋义武,祝长青,等.转基因大豆内外源DNA在少孢根霉发酵中的降解[J].畜牧与兽医,2012,44(12):20-25
[3]韩宗元,江连洲,李杨,等.水酶法提取大豆油的扩大试验研究[J].中国粮油学报,2015,30(2):37-42
[4]Jmln D M,Campbell K,Mahfuz A,et al.Enzyme-Assisted Aqueous Extraction of Oil and Protein from Soybeans and Cream De-emulsification[J].Journal of the American Oil Chemists Society,2008,85(10):985-995
[5]Rosenthal A,Pyle D L,Niranjan K.Aqueous and enzymatic processes for edible oil extraction[J].Enzyme&Microbial Technology,1996,19(6):402-420
[6]Campbell K A,Glatz C E,Johnson L A,et al.Advances in Aqueous Extraction Processing of Soybeans[J].Journal of the American Oil Chemists Society,2011,88(4):449-465
[7]杨柳,江连洲,李杨,等.蛋白酶对水酶法提取大豆油脂及蛋白质的影响研究[J].食品工业科技,2009(10):240-243
[8]王宏立,张祖立,白晓虎.大豆挤压膨化技术及膨化机理的分析[J].农机化研究,2006(1):85-86
[9]Stephanie J,Abdullah M,Devin M.Structure,Protein Interactions and In Vitro Protease Accessibility of Extruded and Pressurized Full-Fat Soybean Flakes[J].Journal of the American Oil Chemists Society,2009,86(5):475-483
[10]Stephanie J,Abdullaha M.Low temperature dry extrusion and highpressure processing prior to enzyme-assisted aqueous extraction of full fat soybean flakes[J].Food Chemistry,2009,114(3):947-954
[11]Li Y,Jiang L Z,Qiao G H.Effect of Extruding Parameters on Soybean Oil Yield of Enzyme-Assisted Aqueous Extraction Processing[J].Advanced Materials Research,2011,148/149:7-18
[12]Bair C W,Snyder H E.Electron microscopy of soybean lipid bodies[J].Journal of the American Oil Chemists Society,1980,57(9):279-282
[13]冯红霞,隋晓楠,齐宝坤,等.挤压膨化参数对水酶法提取大豆油品质的影响[J].中国油脂,2016,41(4):51-55
[14]王心刚,江连洲,李杨,等.真空挤压膨化水酶法提取大豆油的工艺研究[J].中国粮油学报,2013,28(11):28-31
[15]王卫国,朱占齐,吴兴泉,等.微波、超高压处理对转基因大豆外源基因降解的影响[J].中国粮油学报,2015,30(2):20-25
[16]吴洪洪.转基因Roundup Ready大豆外源CP4-EPSPS蛋白及内外源基因在食品加工过程中的降解变化规律[D].南京:南京农业大学,2011
[17]Guan Q,Wang X,Teng D,et al.DNA degradation of genetically modified cottonseed meal during feed processing[J].Applied Biochemistry&Biotechnology,2013,169(2):368-379
[18]王林,韩飞,李爱科,等.不同加工工艺对大豆转基因成分及调控元件的影响[J].大豆科学,2011,30(1):136-140
[19]夏义苗.一种用于提取大豆种子基因组DNA的细胞裂解液及其提取方法:201710888077.0[P].2018-1-12