骆驼蓬(Peganum harmala)蛋白质的分布、提取及活性研究

Distribution,Extraction and Activities of the Proteins from Peganum harmala L.

采用不同溶剂分级提取骆驼蓬(Peganum harmalaL.)根、茎、叶及种子中的蛋白质,结果表明总蛋白以种子中的蛋白含量最高,而营养器官以叶中的蛋白含量较高。当年种子总蛋白含量显著高于贮藏种子。不同溶剂分级提取的当年生种子、根、茎和叶中各组分,以碱提组分蛋白含量最高。用含不同盐离子浓度的缓冲液和不同pH值的广泛缓冲液提取骆驼蓬种子的蛋白质,结果表明优化后的缓冲液条件为含0.2 mol/L NaCl,pH=7.0～8.0的5 mmol/L PBS缓冲液。硫酸铵沉淀法获得的骆驼蓬种子中的蛋白粗提物(1.2 mg/mL)对供试真菌交格链孢菌(Alternaria alternata)、指状青霉菌(Penicillium degitatum)、灰霉病菌(Botrytis cinerea)、稻瘟病菌(Magnaporthe grisea)和意大利青霉菌(Penicillium italicum)等5种植物病原真菌均有抑菌作用,其中对意大利青霉菌和交格链孢菌表现出较好的抑菌活性,抑菌环直径分别为19.50和18.50 mm。对供试细菌表皮葡萄球菌(Staphylococcus epidermidis)、粪肠球菌(Enterococcus faecalis)、臭鼻克雷伯菌(Klebsiella penumoniae)、福氏志贺氏菌(Shigella flexneri)、金黄色葡萄球菌(Staphylococcus aureus)和鲍曼不动杆菌(Acinetobacter baumannii)等6种病原细菌也有抑菌作用,其中对臭鼻克雷伯菌、福氏志贺氏菌、表皮葡萄球菌等病原细菌等有较好的抑制作用,抑菌圈直径分别10.20、10.10和9.30 mm。

The proteins were extracted from the roots ,stems ,leaves and seeds of Peganum harma/a L. by using the different solvents. The results showed that the seeds had the highest content of total protein, while the protein of vegetative organs was mainly distributed in the leaves. The protein content of current growth seeds was significantly higher than that of seeds stored. Among the proteins extracted from the different solvents,the components of total proteins got by NaOH extracting method is the highest percent. The optimized buffer conditions of the protein extraction under the different NaCl concentrations and the different pH buffers were 0.2 mol/L NaCI,pH =7.0-8.0 respectively. The determination of antimicrobial spectrum with the crude protein obtained by sulfate ammonium fractional precipitation indicated that the bacteriostatie action were obvious to both 5 kinds of plant pathogenic fungi such as Ahernaria alternate, Penicillium degitatum,Botrytis cinerea,Magnaporthe grisea and Penicillium italicum,and 6 kinds of pathogenic bacteria such as Staphylococcus epidermidis , Enterococcus faecalis , klebsiella penumoniae , Shigella flexneri ,Staphylococcus aur eus and Acinetobacter baumannii,especially have a distinct antimicrobial activity to two types of fungi,PeniciUium degitatum and Alternaria alternate,with the diameter of inhibition ring was 19.50 and 18.50 mm respectively, and to three types of bacteria, Klebsiella penumoniae,Shigella flexneri and Staphylococcus epidermidis,with the diameter of inhibition ring was 10.20,10.10 and 9.30 mm respectively.