Inactivation Kinetics of Pectin Methylesterase from Peach under High Pressure Carbon Dioxide
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摘要: 采用高压二氧化碳技术(High Pressure Carbon Dioxide, HPCD)处理桃果胶甲基酯酶(Pectin Methylesterase, PME)粗酶液，分析了HPCD对粗酶液中PME的钝化效果及动力学，进一步比较了粗酶液和桃汁两种体系中PME对HPCD的敏感性。HPCD对粗酶液中PME具有较好的钝化效果，处理温度和压力的共同作用导致了PME活性降低，钝化动力学遵从一级动力学模型。随着处理压力和温度的提高，钝化速率逐渐增大，而指数递减时间则逐渐减小；在最优的处理条件(22 MPa、55 ℃)下，HPCD钝酶的钝化速率和指数递减时间分别为0.408 8 min-1和5.63 min。温度为55 ℃时HPCD钝酶的压力敏感指数为16.40 MPa，活化体积为-383.00 cm3/mol；压强为15 MPa时HPCD钝酶的温度敏感指数为13.30 ℃，活化能为1 845.86 kJ/mol。比较HPCD对桃汁和粗酶液中PME的钝化效果，发现HPCD处理16 min后(15 MPa、55 ℃)，桃汁中PME残存酶活在80%左右，而粗酶液中PME活性已完全钝化，表明桃汁体系中存在保护PME活性的因素，有关机制需要进一步研究。Abstract: The inactivation of pectin methylesterase (PME) from peach in buffer and juice by high pressure carbon dioxide (HPCD) was investigated.PME in buffer is effectively inactivated by HPCD, their residual activity decreases with the increase of pressure and temperature.The residual activity of the PME exhibited a fast decrease after a prolonged treatment time; their inactivation kinetics is adequately modeled by the first order kinetics model.With the increase of pressure and temperature, the kinetic rate constant increases and the decimal reduction time decreases for PME in buffer inactivated by HPCD.The Kinetic rate and the decimal reduction time are 0.408 8 min-1 and 5.63 min respectively at the optimal condition (22 MPa, 55 ℃).The activation volume and the pressure sensitivity exponent of inactivation PME by HPCD at 55 ℃ are -383.00 cm3/mol and 16.40 MPa respectively.In contrst, the activation energy and the pressure sensitivity exponent at 15 MPa are 1 845.86 kJ/mol and 13.30 ℃ respectively.The residual activity of PME in peach juice is significantly greater than that in buffer after the same HPCD treatment, indicating that the original PME in the juice is less inactivated by HPCD.
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图 2 HPCD处理装置
Figure 2. High pressure carbon dioxide processing equipment
1.CO2 cylinder; 2.CO2 filter; 3.Pressure gauge; 4.Cooling unit; 5.Plunger pump; 6.Pressure transducer; 7.Biohazard clean safety facility for aseptic operation; 8.High pressure vessel; 9.Thermocouples; 10.Thermostatic bath; 11.Vacuum pump; 12.Displaying panel
图 3 不同压强下HPCD处理(55 ℃)桃PME的钝化动力学
Figure 3. Inactivation kinetics of peach PME by HPCD at 55 ℃ modelled using the first-order kinetic model
图 4 HPCD(55 ℃)钝化桃PME的Zp和Va值理
Figure 4. Zp and Va for pressure-dependence inactivation of peach PME by HPCD at 55 ℃
图 5 不同温度下HPCD处理(15 MPa)桃PME的钝化动力学
Figure 5. Inactivation kinetics of peach PME by HPCD at 15 MPa modelled using the first-order kinetic model
图 6 HPCD处理(15 MPa)钝化桃PME的ZT和Ea值
Figure 6. ZT and Ea for temparature-dependence inactivation of peach PME by HPCD at 15 MPa
图 7 HPCD处理(55 ℃，15 MPa)对桃汁和PME粗酶液的钝化效果(重复次数:n=3；不同字母a, b, c, d, e表示有显著性差异)
Figure 7. Comparison of the AR of peach PME in juice and buffer by HPCD (55 ℃, 15 MPa) (Times of repetition:n=3;Different letters represent that there are significant diffrences)
表 1 不同压强HPCD钝化桃PME的动力学参数(55 ℃)
Table 1. Estimation of inactivation kinetic parameters of peach PME by HPCD at 55 ℃ using the first-order kinetic model
Pressure/(MPa) Kinetic parameters k/(min-1) D/(min) R2(P < 0.05) 5 0.036 5±0.000 8 63.08 0.76 8 0.066 4±0.001 5 34.68 0.79 12 0.168 3±0.004 0 13.68 0.94 15 0.196 5±0.022 2 11.72 0.91 22 0.408 8±0.044 1 5.63 0.86 Note：R2 represents the linear regression coefficient, P represents the significance level. 
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表 2 不同温度HPCD钝化桃PME的动力学参数(15 MPa)
Table 2. Estimation of inactivation kinetic parameters of peach PME by HPCD at 15 MPa using the first-order kinetic model
Temperature/(℃) Kinetic parameters k/(min-1) D/(min) R2(P < 0.05) 25 0.001 2±0.000 5 1 918.82 0.360 35 0.003 6±0.000 8 639.61 0.667 45 0.027 0±0.001 5 85.28 0.970 55 0.196 5±0.022 2 11.72 0.910 Note：R2 represents the linear regression coefficient, P represents the significance level.
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