压力脉动固态发酵微生物蛋白质及机理的研究

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

研究了固态发酵微生物蛋白质的提取和纯化条件,以及压力脉动对固态发酵微生物蛋白质的影响,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,得到的胞内蛋白质的量较多,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蛋白质与从未加周期刺激的微生物中提取的相比,胞内蛋白质的质量提高了34.63%,FPA酶活降低了22.22%,CMCase酶活降低了38.65%,而胞外蛋白质的质量、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6 d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,减少了分子量约为80400的组分,但增加了分子量约为28520的组分。

The effect of varying the extraction and purification conditions for the microorganism protein obtained from solid state fermentation and the effect of periodical dynamic changes of air on the protein have been studied. The mechanism of solid-state fermentation with periodical dynamic changes of air is also discussed. The results show that when Tris-HC1 buffer was added to the solid-state fermentation system, the maximum amount of intracellular protein. The recovery ratio of FPA activity is 83.6 %, while the recovery ratio of CMCase activity is 67%. Compared with static solid state fermentation, periodic dynamic changes of air afford a higher protein mass; from ca. 1 g of fermentation microorganism, 5.3 mg of intracellular protein （an increase of 34.6% ） and 9.09 mg of extracellular protein （an increase of 17.8% ） were obtained on the sixth day of the fermentation. The FPA enzyme activity and CMCase enzyme activity of the extracellular protein are 1.739μmol/s and 109. 592 μmol/s respectively, which represent increases of 60.1% and 21.2 % over the corresponding values for static solid state fermentation. The FPA enzyme activity and CMCase enzyme activity of the intracellular protein are 0. 245 μmol/s and 6. 392 μmol/s respectively, which represent decreases of 22.2 % and 38.7 % over the corresponding values for static solid state fermentation. The enzyme activity of the microorganism extracellular protein in solid-state fermentation with periodic dynamic changes of air on the fifth dayis nearly equal to that on the sixth day without periodic dynamic changes of air, so that the period of fermentation can be shortened. The SDS-PAGE experiments suggest that pulsating air pressure stimulation leads to a decrease in the amount of protein component with relative molecule mass of about 80400, and an increase in the amount of the the protein component with relative molecular mass of about 28520.