The Metabolic Regulation Of Sulforaphan In Vitro Cell From Brassica Oleracea Var. Botrytis

Sulforaphane can be obtained form cruciferous vegetables as one of the products of enzymatic or acid hydrolysis of glucoraphanin, which has been indentified the best anticancer activity and is also the strongest Phaseâ…¡enzyme inducer. Molecular weight is 177.29, formula is C₆H₁₁SNO。This study established induction and culture system of broccoli callus. Weighing method for growth,organic solvent method for extraction and HPLC method for determination were used to study the influences of supplementary materials, light quality and harvest time on the growth of the callus and its sulforaphane accumulation. The results showed that:1.The best induction and subculture media of broccoli callus were MS+1.0mg·L^-12,4-D+2.0mg·L^-16-BA and MS+1.0mg·L^-1NAA+2.0mg·L^-16-BA. Root was the best organs to induce broccoli callus and callus induced from root had the best multiplication. 2.The best extraction conditions of sinigrin were chosen by unifactor experiment.Extraction agent:dichloromethane. Extraction temperature:30℃. Extraction time: 2hour. Extraction times:2 times. Material to solvent ratio:20:1. The yield of sulforaphane had significantly increased after enzymatic hydrolysis.3.The light quallties for the multiplication were in the order of white>red and blue>green>yellow. Different light had different effects on soluble protein content and water content. Red and blue light had higher soluble protein and water content.They have correlation with multiplication. The red and blue light promoted the accumulation of sulforaphen, the content reached 747.33μg·g^-1,1.38 times higher than that of white. Sulforaphen production of red and blue light reached the maximum with 260.47μg·flask^-1, 1.03 times more than the white.4.The additions of five kinds amino acids promoted the broccoli callus multiplication. When the concentrations of glycine, phenylalanine and methionine were respectively 10mg/L,30mg/L and 30mg/L,the sulforaphane contents of broccoli callus were promoted obviously.Leucine did not promote the sulforaphane content and glutamate inhibitioned the sulforaphane content.5.In the growth cycle of broccoli callus, the multiplication growth of callus showed S-type curve and the content of sulforaphane was bimodal curve,which reached the maximum 867.34μg·g^-1 in 18d.According to the gross production, the best harvest time was 27d,the sulforaphen production reached the maximum 331.19μg·flask^-1.