Research Of Eucommia Male Flower Protein’s Separation And Purification

The Pollen of Eucommia ulmoides is determined by the male flower from eucommia ulmoides family, so it is called eucommia staminate flower pollen. Early research on eucommia is mainly confined to the eucommia ulmoides Oliv and its leaves, and the mainly contents are small molecule compounds. Since the late 70’s, a fanatical tide on pollen studies appear at home and abroad. Many scholars have done a lot of research work. But research and development on eucommia flower pollen at home and abroad are rare reported.In this study, we isolated and purified bioactive proteins from eucommia staminate flower pollen which sets foundations to the development and utilization of eucommia ulmoides pollen in the healthcare or medical industry. This study on the experiment of eucommia ulmoides pollen for materials, optimized from single factor conditions by breaking experiment, ultrasonic extraction power, time, frequency, solid-liquid ratio, and buffers p H and so on. Extractized the biologically active protein components from eucommia staminate flower pollen by salting out, dialysis, dextrin gel column chromatography and high performance liquid chromatography method, and determination of protein content and the ultraviolet wavelength scanning analysis, the main results are as follows:1. Take eucommia ulmoides pollen ratio and corresponding albumin yields as the index to mixed manual grinding temperature, liquid nitrogen, and liquid nitrogen mixture ball mill grinding, we compared the three methods, and then find that pollen derived nitrogen mixture ball mill grinding method is the more simple and efficient method of breaking under laboratory conditions.2. Valley of alkali-soluble protein is the major constituent in eucommia staminate flower pollen proteins, which accounts for 80% percent of total protein extraction of eucommia ulmoides pollen classification.3.Through the determination of classification Eucommia ulmoides male flower pollenprotein extraction rate, we obtained that the grading of ultrasonic cell disruption with instrument-assisted extraction of total protein extraction rate is 72.22%, namely the extraction of 1.9 times ultrasonic cleaning machine-assisted extraction of 3.2 times.4. The analysis of single-factor experimental determined the optimum ultrasonic extraction of eucommia ulmoides pollen conditions are: solution p H 10, 1:25 solid-liquid ratio, ultrasonic power 300 W, ultrasonic 10 min, and ultrasonic frequency 4 times.5.In the process of ammonium sulfate fractionl precipitation, through various experiments of saturation, we determined six of the precipitate components are 10%, 30%, 45%, 60%, 75%, 90%. 30% of which proteins are major components of the total precipitation, accounting for more than 90%, so took 30% part of the test as the main component of the experiment.6. After twice Sephadex G-50 chromatography, we got two relatively pure components. Identify the two components through HPLC method,the components have at least five more protein.7. Through the outside dialysis fluid of ultraviolet wavelength scanning, it displayed in 200~225nm far ultraviolet wavelengths of light absorbed, which suggests that there are small molecular weight proteins in the outside dialysis fluid.