压力脉动固态发酵微生物蛋白质及机理的初步研究

研究了固态发酵微生物蛋白质的提取、纯化条件以及压力脉动对固态发酵微生物蛋白质的影响，并初步探讨了压力脉动固态发酵的作用机理。结果表明在发酵后的酶曲中，加入Tris-HCl提取液，可得胞内蛋白质，FPA酶活与CMCase酶活回收率分别为83.6%与67%,纯化效果较好。从压力脉动外界周期刺激固态发酵干酶曲中提取的胞内蛋白质与从未加周期刺激的微生物中提取的相比，胞外蛋白质的质量、FPA酶活和CMCase酶活分别提高了17.75%、60.08%和21.17%。压力脉动固态发酵5d的微生物胞外蛋白质的酶活,与静态固态发酵6d的相当，发酵周期缩短。压力脉动外界周期刺激使蛋白质组分有所变化，减少了相对分子质量约为80400的组分，但增加了相对分子质量约为28520的组分。

Study of microorganism protein and mechanism in solid state fermentation with periodical dynamic changes of air

The extracting and purifying conditions for microorganism protein in Solid State Fermentation and the effect of periodically dynamic changes of air on protein were studied；The mechanism on solid-state fermentation with periodically dynamic changes of air was simply discussed. Results show that three milliliters Tris-HCl buffer added to the solid-state fermentation microorganism about one gram to extract intracellular protein, from which got the most intracellular protein about 3.828mg. That the impure protein is deposited with three times acetone in -20℃for 2h produce the highest purification, native protein and Clear sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) pattern. The recovery rate of FPA activity is 83.6% ,while the recovery rate of CMCase activity is 67%.Contrast to the period of static solid state fermentation, The periodically dynamic changes of air get a higher protein mass number from fermentation microorganism about one gram , which of intracellular protein increasing 34.63% reach 5.3mg and of extracellular protein increasing 17.75% reach 9.09mg in sixth day of the fermentation, enhance on enzyme activity of extracellular protein which of FPA enzyme activity and CMCase enzyme activity respectively are 1.739μmol/s and 109.592μmol/s ,increasing 60.08% and 21.17%，but reduce litter on enzyme activity of intracellular protein which of FPA enzyme activity and CMCase enzyme activity respectively are 0.245μmol/s、6.392μmol/s, reducing 22.22% and 38.65%. The enzyme activity of microorganism extracellular protein in solid-state fermentation with periodically dynamic changes of air in fifth day is nearly equal to that without periodically dynamic changes of air in sixth day, so that period of fermentation is shortened. The SDS-PAGE suggest air press pulsating stimulation increase protein component whose relative molecule mass is about 80400,while disappear one protein component about 28520.