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目的对抗冻蛋白Tm AFP84进行理化性质分析。方法对总蛋白质进行浓缩,用Bradford法测定Tm AFP84蛋白量,并对Tm AFP84蛋白的酸碱稳定性和热稳定性进行研究。结果 Tm AFP84蛋白经过100℃热处理后,其低温保护效果和未经热处理的Tm AFP84蛋白保护效果相比下降了约20%,但仍能使大肠杆菌在0℃的相对存活率达1.0%左右,显著高于对照水处理的大肠杆菌存活率(P<0.05)。但是100℃处理5min和10min二者间Tm AFP84蛋白的保护大肠杆菌的效果差异不显著(P>0.05)。Tm AFP84蛋白经过不同酸碱度p H1、p H3、p H12的缓冲液处理后,低温保护能力有不同程度下降。p H1液处理下降22%、p H3液处理下降12%、p H12液处理下降17%左右,但经p H14液处理5d后大肠杆菌抗冻保护效果下降最大,其存活率几乎为零。结论实验证明Tm AFP84在较广的p H范围和较宽的温度范围内具有较强的稳定性,这为扩大生产抗冻蛋白Tm AFP84及深入研究其性质和应用奠定了基础。
Objective To analyze the physicochemical properties of anti-Tm AFP84. Methods Total protein was concentrated. The amount of Tm AFP84 protein was determined by Bradford method. The acid-base stability and thermal stability of Tm AFP84 protein were also studied. Results After heat treatment at 100 ℃ for Tm AFP84 protein, the protective effect of Tm AFP84 protein decreased by about 20% compared with that of Tm AFP84 without heat treatment. However, the relative survival rate of Tm AFP84 protein at 0 ℃ was about 1.0% Was significantly higher than that of control water-treated E. coli (P <0.05). However, there was no significant difference (P> 0.05) in the effect of Tm AFP84 protein protection E. coli at 100 ℃ for 5min and 10min respectively. Tm AFP84 protein after different pH p H1, p H3, p H12 buffer treatment, low temperature protection ability to varying degrees decline. pH1 treatment decreased by 22%, pH3 treatment decreased by 12%, pH12 treatment decreased by about 17%, but after treated with pH14 solution for 5 days, Escherichia coli decreased the most, and its survival rate was almost zero. Conclusions The experiment proves that Tm AFP84 has strong stability in a wide range of p H and a wide temperature range, which lays the foundation for expanding the production of anti-freeze protein Tm AFP84 and further studying its properties and applications.