渔业科学进展Issue(4)：132-140,9.DOI:10.11758/yykxjz.20140420
响应面法优化丹参中抗无乳链球菌活性成分的提取工艺
Optimization of the Extraction Conditions of Active Components for Inhibition of Streptococcus agalactiae from Salvia miltiorrhiza Bge. Using Response Surface Methodology
摘要
Abstract
In recent yearsStreptococcus agalactiae(SA) has become the most harmful pathogen to cultured tilapia in Hainan and many southern provinces of China.Salvia miltiorrhizaBge is highly resistant to SA, but it contains complex components and which component is responsible for the resistance to SA has not been reported. Here we investigated the effects of extraction conditions on the yield of anti-SA components inS. miltiorrhizaBge and optimized the extraction method. We first developed the agar diffusion method for the quantitative analysis of extracted anti-SA components; then we used the yields of anti-SA components fromSalvia miltiorrhizaBge as the responsive values to study the effects of extraction conditions on the yield of the anti-SA components, including the extraction solvent, extraction times, the ratio of liquid to solid, infiltration time, temperature and extraction time in single-factor experiments; finally we further optimized the ratio of liquid to solid, infiltration time, temperature and extraction time using the four-factor-three-level Box-Behnken experimental design with response surface method. The results of the agar diffusion showed that the standard equation for the determination of anti-SA components fromS. miltiorrhizaBge wasY= 4.842X+ 16.707 (R2= 0.9873). The results of the single-factor experiments showed that the most suitable extraction solvent and times was 20% ethanol aqueous solution for 1 time. The results of the Box-Behnken experimental design with response surface method showed that the infiltration time, the extraction temperature, and the ratio of liquid to solid had extremely vital effects on the responsive values. The optimum extraction conditions were as follows: the ratio of liquid to solid was 37.8:1.0 (ml:g), the infiltration time was 104 min, the extraction temperature was 86.7℃ and the extraction time was 182 min. The predicted maximum yield of anti-SA components was 25.95%, under the RSM model-generated optimal extraction conditions. The results of four parallel validation experiments showed that the actual yield of anti-SAcomponents was (25.87 ±0.03)%, and the relative error between predicted and actual values was 0.31%. The yield of anti-SA components was 2.59 times higher than before. These demonstrated that our optimization method was efficient and reliable. Furthermore, our study provided important clues to future research on the anti-SA components ofS. miltiorrhizaBge, and significantly contributed to the safe and efficient control of diseases caused byS. agalactiae.关键词
无乳链球菌/丹参/提取工艺/响应面法Key words
Streptococcus agalactiae/Salvia miltiorrhizaBge/Extraction conditions/Response surface methodology分类
农业科技引用本文复制引用
邓恒为,郭伟良,孙晓飞,谢珍玉,王世锋,胡文婷,周永灿..响应面法优化丹参中抗无乳链球菌活性成分的提取工艺[J].渔业科学进展,2014,(4):132-140,9.基金项目
海南省重点科技计划项目(ZDXM20120005)、海口市重点科技项目、海南大学地方服务项目和海南大学青年基金(qnjj1206)共同资助。 （ZDXM20120005）
