植物乳杆菌lp15-2-1转化合成共轭亚油酸的研究
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摘要
共轭亚油酸(Conj ugated Linoleic Acid,简称CLA)是一系列含有共轭双键、具有位置和构象异构的十八碳二烯酸的总称,是必需脂肪酸亚油酸(Linoleic Acid.,简称LA)的一组异构体。CLA具有抗氧化、抗癌、抗动脉粥样硬化、减少机体脂肪等功能。本论文主要研究一株从泡菜中筛选获得的植物乳杆菌(Lactobacillus plantarum)lp15-2-1转化LA为CLA的能力,并初步探讨了亚油酸异构酶(Linoleic acid isomerase,简称LAI)的酶学性质及定位,为后续深入研究和大规模工业化生产CLA提供试验依据。
研究了植物乳杆菌1p15-2-1生长细胞转化LA合成CLA的反应条件。在单因素试验的基础上,采用响应面Box-Benhnken试验设计,建立初始pH值、培养温度和LA浓度的二次多项式回归方程模型,所得最佳参数：温度30℃,LA浓度0.21mg/mL,初始pH6.3,CLA实际产量为44.77μg/mL,转化率为21.32%,与优化前转化率7.78%相比,有了很大提高。
研究了植物乳杆菌lp15-2-1静息细胞转化LA合成CLA的反应条件。在单因素试验的基础上,采用中心组合设计,建立培养温度、LA浓度、pH值、培养时间、细胞浓度的二次多项式回归方程模型,所得最佳参数：温度31℃,LA浓度3.8mg/mL,初始pH5.7,培养时间84h,细胞浓度32%, CLA实际产量为354.58μg/mL,与优化前产量相比,有了很大提高。
研究了不同细胞破碎方式对植物乳杆菌1p15-2-1产CLA的影响,初步探讨了LAI的酶学性质,测得LAI的最适反应温度为30℃、最适pH值为5.5,该酶在弱酸性条件下活性较高。并对该菌株LAI的定位情况进行了研究,差速离心试验结果表明LAI是膜酶,即结合在细胞膜上的酶,而去垢剂Trtion X-100的试验效果不显著。
Conjugated linoleic acid(short for CLA) is a set of octadecadienoic acid with conjugated double bounds, and they're isomers to essential fatty acids linoleic acid(short for LA). It's reported that CLA has a function of anti-oxidant, anti-cancer, anti-atherosclerosis and reduce body fat and so on. In this paper, we researched the ability of transforming LA into CLA by Lactobacillus plantarum lp15-2-1, and discussed LA isomerase's enzymatic properties, to provide experimental basis for subsequent in-depth study and large-scale industrial production.
The condition of converting LA into CLA was studied by using Lactobacillus plantarum lp15-2-1. On the basis of single factor experiments, designed a quadratic polynomial regression model by using Box-Benhnken trial design. Results showed that the optimal parameters was:incubation temperature 30℃, the initial level of LA 0.21 mg/mL, pH 6.3, the actual yield of CLA is 44.77μg/mL, the conversion rate is 21.32%, compare to 7.78% at the beginning.
The condition of converting LA to CLA was studied by washed cells from Lactobacillus plantarum lp15-2-1. On the basis of single factors, central composite design was used to make a quadratic polynomial regression model. And the results showed that the optimal parameters was:incubation temperature 31℃, the initial level of LA 3.8mg/mL, pH 5.7, culture time 84h and the cell concentration 32%, the actual yield of CLA is 354.58μg/mL.
Different cell disruption methods on Lactobacillus plantarum lpl 5-2-1 producing CLA was researched, discussed LA isomerase's enzymatic properties, and the best conditions was that 30℃, pH5.5. Plus, it's more active in slightly acidic environment. The localization of LAI was studied by differential certrifugation which showed that LAI was a membrane-bound enzyme, while adding detergent Triton X-100 didn't show significant results.
研究了植物乳杆菌1p15-2-1生长细胞转化LA合成CLA的反应条件。在单因素试验的基础上,采用响应面Box-Benhnken试验设计,建立初始pH值、培养温度和LA浓度的二次多项式回归方程模型,所得最佳参数：温度30℃,LA浓度0.21mg/mL,初始pH6.3,CLA实际产量为44.77μg/mL,转化率为21.32%,与优化前转化率7.78%相比,有了很大提高。
研究了植物乳杆菌lp15-2-1静息细胞转化LA合成CLA的反应条件。在单因素试验的基础上,采用中心组合设计,建立培养温度、LA浓度、pH值、培养时间、细胞浓度的二次多项式回归方程模型,所得最佳参数：温度31℃,LA浓度3.8mg/mL,初始pH5.7,培养时间84h,细胞浓度32%, CLA实际产量为354.58μg/mL,与优化前产量相比,有了很大提高。
研究了不同细胞破碎方式对植物乳杆菌1p15-2-1产CLA的影响,初步探讨了LAI的酶学性质,测得LAI的最适反应温度为30℃、最适pH值为5.5,该酶在弱酸性条件下活性较高。并对该菌株LAI的定位情况进行了研究,差速离心试验结果表明LAI是膜酶,即结合在细胞膜上的酶,而去垢剂Trtion X-100的试验效果不显著。
Conjugated linoleic acid(short for CLA) is a set of octadecadienoic acid with conjugated double bounds, and they're isomers to essential fatty acids linoleic acid(short for LA). It's reported that CLA has a function of anti-oxidant, anti-cancer, anti-atherosclerosis and reduce body fat and so on. In this paper, we researched the ability of transforming LA into CLA by Lactobacillus plantarum lp15-2-1, and discussed LA isomerase's enzymatic properties, to provide experimental basis for subsequent in-depth study and large-scale industrial production.
The condition of converting LA into CLA was studied by using Lactobacillus plantarum lp15-2-1. On the basis of single factor experiments, designed a quadratic polynomial regression model by using Box-Benhnken trial design. Results showed that the optimal parameters was:incubation temperature 30℃, the initial level of LA 0.21 mg/mL, pH 6.3, the actual yield of CLA is 44.77μg/mL, the conversion rate is 21.32%, compare to 7.78% at the beginning.
The condition of converting LA to CLA was studied by washed cells from Lactobacillus plantarum lp15-2-1. On the basis of single factors, central composite design was used to make a quadratic polynomial regression model. And the results showed that the optimal parameters was:incubation temperature 31℃, the initial level of LA 3.8mg/mL, pH 5.7, culture time 84h and the cell concentration 32%, the actual yield of CLA is 354.58μg/mL.
Different cell disruption methods on Lactobacillus plantarum lpl 5-2-1 producing CLA was researched, discussed LA isomerase's enzymatic properties, and the best conditions was that 30℃, pH5.5. Plus, it's more active in slightly acidic environment. The localization of LAI was studied by differential certrifugation which showed that LAI was a membrane-bound enzyme, while adding detergent Triton X-100 didn't show significant results.
引文
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