预测微生物学在高压二氧化碳杀菌动力学的应用进展

高压二氧化碳(HPCD)技术是目前越来越受关注的非热杀菌技术之一。目前有关HPCD杀菌预测模型的研究报道较少。利用食品预测微生物学方法和理论,通过研究各个因素对HPCD杀菌过程的影响,建立预测模型和回归方程,可以更好的理解HPCD杀菌动力学及机理,提高其杀菌效果,确保食品安全。本文就食品预测微生物学在HPCD杀菌动力学研究中的应用进行综述分析,并提出未来可能的研究方向。      

高压二氧化碳杀菌机理研究进展

高压二氧化碳（HPCD）技术作为一种新型非热杀菌技术,越来越受到研究者的关注,但目前对其杀菌机理尚未明确。本文分析了目前国内外有关HPCD杀菌机理的研究进展,并分析了HPCD对微生物细胞内几个作用靶点包括细胞壁、细胞膜、细胞质、蛋白质、酶和核质的影响,以期为进一步研究和应用提供理论依据。     

高压二氧化碳对鲜榨梨汁杀菌效果及动力学研究

本文研究了高压二氧化碳(HPCD)对梨汁中细菌菌落总数的影响,并分析其杀菌动力学。结果表明:随着温度、压强升高以及处理时间延长,梨汁中细菌菌落总数显著降低(p<0.05);在相同温度和处理时间条件下,HPCD处理显著高于热处理的杀菌效果,处理温度对HPCD杀菌具有协同效应;当HPCD处理条件为30MPa、40℃、60min时,灭菌效果最佳,梨汁中细菌菌落总数的残存率降低了2.66个对数;Weibull模型能较好地拟合HPCD处理后梨汁中细菌菌落总数的失活曲线,模型动力学参数比例因子a和形状因子b随压力增加和温度升高呈现逐渐变小的规律性变化。      

Mathematical modeling of Saccharomyces cerevisiae inactivation under high‐pressure carbon dioxide

High‐pressure carbon dioxide inactivation curves of Saccharomyces cerevisiae at different temperatures were analysed using the modified Gompertz model. Comparable λ and μ values were obtained under pressure treatment as function of temperature. The phase of disappearance (λ) and the inactivation rate (μ) of S. cerevisiae were inversely related. Higher μ values were obtained at 50°C than at 40, 30, and 20°C under 10.0 MPa CO₂ pressure. Increased pressure and temperature had significant effects on the survival of S. cerevisiae. Arrhenius, linear and square‐root models were used to analyse the temperature dependence of the inactivation rate constant. For the Arrhenius model the activation energy (E_μ) was 56.49 kJ/mol at 10.0 MPa, and 55.70, 53.83, and 52.20 kJ/mol at 7.5, 5.0, and 2.5 MPa, respectively. Results of this study enable the prediction of yeast inactivation exposed to different CO₂ pressures and temperatures.     

高压二氧化碳对西瓜汁中过氧化物酶钝化动力学的研究

为了延长西瓜汁的货架期,研究高压二氧化碳处理对西瓜汁中过氧化物酶(peroxidase,POD)的钝化效果。研究以热处理为对照,比较不同高压二氧化碳处理温度和压强对其POD活性残存率的影响。采用二段式模型模拟POD的钝化效果(R2>0.975),证明西瓜汁中POD存在敏感型和稳定型两部分。通过动力学分析考察两种类型POD对温度和压强的敏感性,确认高压二氧化碳处理温度、压力、时间、pH和CO2等因素共同导致了POD活性的降低。高压二氧化碳处理比热处理可以更加有效钝化西瓜汁中POD,有助于延长西瓜汁的货架期。     

Inactivation of polyphenol oxidase from frozen red raspberry (Rubus idaeus L.) by high pressure carbon dioxide treatment

The effect of high pressure carbon dioxide (HPCD) treatment on polyphenol oxidase (PPO) from frozen red raspberry (Rubus idaeus L.) was evaluated. Moreover, the inactivation kinetics of its PPO was simulated by first‐order reaction theory. The minimum of PPO residual rate was 36.6% under 30 MPa and 55 °C for 60 min by HPCD treatment, while that was 66.8% at 55 °C for 60 min by thermal treatment. Moreover, the decimal reduction time of PPO decreased rapidly after HPCD treatment, compared to that of the thermal treatment. The thermal treatment at 55 °C takes a similar time to reach 10% PPO residual rate with HPCD treatment under 30 MPa at 35 °C. One reason for the results was that activation energy of PPO reduced from 98.9 to 14.6 kJ mol^?1 after HPCD treatment. Therefore, HPCD treatment showed stronger capacity to inactivate PPO from frozen red raspberry than the thermal treatment at same temperature.     

高压CO2与超高压均质协同杀菌装置的研发

高压CO_2杀菌和超高压均质杀菌都是非常有前途的非热杀菌技术。为了进一步提升其杀菌能力,把高压CO_2杀菌与超高压均质杀菌有机地结合起来,构建一个新型的高压CO_2与超高压均质协同杀菌技术,并对其工作原理、工作特点、结构组成进行系统的阐述,搭建协同杀菌试验平台。通过酿酒酵母的杀菌试验,验证了协同杀菌系统优于单一的高压CO_2杀菌或超高压均质杀菌。     

A comparative study of inactivation of peach polyphenol oxidase and carrot polyphenol oxidase induced by high‐pressure carbon dioxide

The inactivation of polyphenol oxidase (PPO) in peach juice and PPO in carrot juice was investigated by high‐pressure carbon dioxide (HPCD), and their inactivation kinetics was analysed and compared. The temperature was 35–55 °C, the pressure was 5–15 MPa under HPCD condition. Results showed that HPCD enhanced the inactivation effect of the temperature on the two PPOs. The inactivation kinetics of peach PPO was well fitted to a first‐order kinetic model, of carrot PPO to a fraction‐conversion model as a function of temperatures or pressures. Susceptibility of the rate constant k of peach PPO was not altered and of carrot PPO was lessened to the temperature, but the susceptibility of the rate constant k of peach PPO and carrot PPO to the pressure was not changed when the pressure was >8 or 12 MPa, indicating the presence of a threshold pressure.     

高压CO_2对树莓汁品质的影响

研究了高压二氧化碳(High Pressure Carbon Dioxide,HPCD)对树莓汁品质的影响。处理条件为:压强10、20、30MPa,温度35、45、55℃,时间15、30、45、60min。测定其浊度、褐变指数、悬浮稳定性、pH、可溶性固形物含量和色泽。处理后样品褐变指数降低,浊度和悬浮稳定性增加,pH、可溶性固形物含量和色泽没有显著变化。     

猪肉超氧化物歧化酶超高压失活动力学研究

以猪背最长肌为材料,用不同压力(400~750 MPa)结合热(20~60℃)处理,以研究猪肉中超氧化物歧化酶(SOD)的超高压失活动力学情况。结果表明:在某一恒定的温度下(20℃≤T≤60℃),猪肉中SOD的失活速率常数k随着处理压力的增加而增大。各温度条件下的反应活化体积Va都是负值,表明随着压力的增加SOD失活速率常数在增大。SOD反应活化体积Va的绝对数值在40℃及60℃时达到最小,表明此温度下SOD超高压失活速率常数受压力影响最小,具有较高的压力稳定性。在实验压力范围之内(400 MPa≤P≤750 MPa),反应活化能Ea值总体呈减小趋势,说明随着压力的增加,温度对SOD失活速率常数的影响在减弱。      

The use of carbon dioxide at high pressure to control nine stored-product pests

The development of alternative treatments for pest control in food commodities is an increasing demand from the food industry, which should meet consumer demands for the reduced use or elimination of pesticides. The use of carbon dioxide (CO₂) at high pressure is one of the most rapid options for arthropod pest control among current commercial treatments, offering complete control within hours. The present study aimed to establish the efficacy of the technique against different stages of several insect and mite pests that affect stored-food products. Standard food diets containing eggs, larvae, pupae and adults of Oryzaephilus surinamensis, Cryptolestes ferrugineus, Lasioderma serricorne, Sitophilus oryzae, Rhyzopertha dominica, Acanthoscelides obtectus, Ephestia kuehniella, Liposcelis bostrychophila and Tyrophagus putrescentiae were treated with CO₂ for different times at two high pressures, 15 and 20 bar. A high level of control was achieved for most species and development stages when they were treated with CO₂ at 20 bar for 60 min. However, the efficacy at 15 bar was much lower. The eggs of L. bostrychophila and L. serricorne showed the highest tolerance of all species/stages and survived the most extreme conditions tested. The beetle O. surinamensis, the moth E. kuehniella, and the mite T. putrescentiae were easier to kill than the other species tested. Our results confirmed that the use of high-pressure CO₂ offers an effective and fast way to control most stored-product pests that affect food commodities.     

响应曲面法建立乳酸菌高压二氧化碳杀菌模型

采用三因素三水平的Box-Behnken设计,利用响应曲面法（RSM）对影响高压二氧化碳（HPCD）杀菌效果的主要因素（温度、压力和时间）进行研究,以杀灭乳酸菌的对数值为响应函数,建立了猪肉糜中乳酸菌的HPCD杀菌模型。对模型进行变异分析、决定系数分析、t值检验、共线性分析和显著性因子对杀菌效果影响的分析结果表明,采用二阶效应模式构建的模型显著性好、准确度高;显著因子时间、压力的二次方、温度-压力交互项、压力、温度对模型的重要性依次递增且其独立性可靠;温度、压力和时间均能增强HPCD对乳酸菌的杀菌效果。      

超高压对双孢蘑菇的杀菌效果和动力学的研究

以细菌总数、大肠菌群、酵母菌和霉菌数为对象,研究了超高压(High hydrostatic pressure,HHP)处理对双孢蘑菇(Agaricus bisporus)的杀菌效果和杀菌动力学。双孢蘑菇在300、400、500、600MPa压力下,室温下分别用HHP处理2.5～25min。结果表明:随压力的升高和时间的延长,杀菌效果增强;霉菌、酵母对压力较为敏感,400MPa处理2.5min可将其全部杀死;300MPa处理2.5min可完全杀灭双孢蘑菇中的大肠菌群。应用Weibull模型对不同处理条件下双孢蘑菇的杀菌效果进行拟合,拟合动力学曲线的决定系数R2均大于0.97,拟合效果较好。提出了双孢蘑菇的HHP杀菌的最优杀菌工艺参数,即600MPa处理5min,该条件即可以有效杀灭双孢蘑菇中的微生物。     

Combined thermal and high pressure inactivation kinetics of tomato lipoxygenase

The combined isothermal (10–60 °C) and isobaric (0.1–650 MPa) inactivation kinetics of lipoxygenase (LOX) extracted from tomatoes and reconstituted in a tomato purée were studied. Thermal inactivation of LOX at atmospheric pressure proceeded in the temperature range of 45–65 °C. LOX inactivation did not follow first order kinetics; the data could be fitted assuming that the two isoforms of LOX with different thermostability were present. Combined thermal and high pressure inactivation occurs at pressures in the range of 100–650 MPa combined with temperatures from 10–60 °C, and followed first-order kinetics. In the high-temperature/low-pressure range, (T≥50 °C and P≤300 MPa) an antagonistic effect is observed, therefore, the Arrhenius and Eyring equation cannot be used over the entire temperature and pressure range. Small temperature dependence is found in the low-temperature/high pressure range. A third degree polynomial model was successfully applied to describe the temperature–pressure dependence of the inactivation rate constants, which can be useful to predict inactivation rate constants of tomato LOX reconstituted in tomato purée in the temperature–pressure range studied.     

高压CO2对水酶法乳状液破乳影响的研究

研究高压CO2破乳过程中的各种因素对破乳效果的影响,并采用透射电镜来观察乳状液破乳前后油脂和蛋白的变化情况。在单因素基础上通过响应面优化得到最佳破乳条件:高压CO2压力为13.96MPa,温度155.64℃,处理时间21.46 s,浓度0.53 g/cm3,破乳率最优值(94.88±0.30)%。并且通过透射电镜观察发现,破乳前乳状液中大部分油脂被蛋白牢牢包裹,然而破乳后乳状液中包裹油脂的蛋白被破坏,油脂释放并结合在一起。     

High pressure carbon dioxide treatment for fresh-cut carrot slices

The effects of high pressure carbon dioxide (HPCD) treatment on natural microorganisms, indigenous enzyme activity, damage to cell membranes and hardness in fresh-cut carrot slices were investigated. 1.86 log₁₀ cycle reduction for aerobic bacteria (AB) and 1.25 for yeasts and molds (Y&M) were achieved at 5 MPa and 20 °C for 20 min. The residual activity (RA) of peroxidase (POD), polyphenol oxidase (PPO), and pectinmethylesterase (PME) exhibited initially increase and secondly decrease with treatment time and their minimum activity was 75.8%, 90.9% and 52.8% at 5 MPa and 20 C for 15 min, respectively. Membrane damage was evaluated by relative electrolyte leakage (REL) and malondialdehyde (MDA) content. HPCD caused a significant increase of REL in carrot slices and the REL of carrot slices treated at 5 MPa and 20 °C for 15 min was 5.7 times as much as that of the untreated, however, HPCD showed no effect on MDA content. The hardness was well retained after HPCD treatment and the largest loss was 7.9% at 5 MPa and 20 °C for 15 min.

Industrial relevance

Fresh-cut carrot slices are one of the most widely used products in prepared salads, and it required strict treatment conditions to protect its quality, especially to prevent microbial spoilage and enzymatic discoloration. HPCD is one promising novel non-thermal technique without compromising the flavor, taste and nutrition aspect of food. This study analyzed the effectiveness of HPCD as a method of preserving fresh-cut carrot slices, including inactivating natural microorganisms and enzymes which are crucial to quality control. Available data provided in this study will benefit the fresh-cut fruits and vegetables industry.     

高压二氧化碳处理对牛通脊颜色和肌红蛋白的影响

在35℃、30min,不同压力(0、7、14、21、28、35MPa)和在35℃、21MPa,不同时间(0、10、20、30、40、50min)对牛通脊进行高压二氧化碳(HPCD)处理,测定肉的色泽和肌红蛋白(Mb)指标的变化。结果表明:HPCD处理能使肉由红色逐渐变成灰棕色,显著提高L*值(p<0.05)、降低a*值(p<0.01)。在35℃、30min条件下,7、14、21MPa的处理分别使L*值提高4.891、9.494、16.432;a*值降低1.575、3.573、5.872。同时,HPCD处理显著降低了肌红蛋白的总量(p<0.01),7～35MPa处理后分别降低了0.034、0.048、0.054、0.044、0.061mmol/L/L。另外,提高压力、延长时间也能显著降低高铁肌红蛋白(MetMb)比例(p<0.05),提高氧合肌红蛋白(MbO2)比例(p<0.01)。4℃冷藏7d后,与对照组相比,HPCD处理均能提高肌红蛋白的稳定性。该研究为HPCD技术在肉类中的应用提供理论参考。     

高压二氧化碳保鲜双孢蘑菇的工艺优化

为了优化高压二氧化碳(high pressure carbon dioxide,HPCD)保鲜双孢蘑菇的工艺,在单因素实验基础上,采用响应面法分析HPCD处理压力、处理时间和处理温度对双孢蘑菇硬度和颜色的影响并对其进行优化。结果表明,HPCD保鲜双孢蘑菇的最佳工艺条件为:处理压力0.3 MPa,处理时间3 min,处理温度17℃;与对照相比,该条件HPCD处理后的双孢蘑菇可以更好的保持其硬度和颜色,低温贮藏8 d后的硬度为191.7 N(提高了18.85%),褐变指数(BI)为33.13(降低了16.80%)。因此,HPCD技术可以应用于双孢蘑菇的贮藏保鲜,具有良好的前景。      

Effect of carbon dioxide on the inactivation of florida spiny lobster polyphenol oxidase

Florida spiny lobster (Panulirus argus) polyphenol oxidase (PPO) exposed to CO₂ (1 atm) at 33, 38, and 43°C showed a decrease in enzyme activity with increased heating time. Enzyme inactivation by CO₂ was faster for PPO heated at 43 than at 33 or 38°C. Inactivation kinetics revealed PPO was more labile to CO₂ and heat in the range 33–43°C than to heat alone. Kinetic data also revealed that CO₂, in addition to pH changes, was involved in the loss of PPO activity. Studies using get electrophoresis showed no differences in protein patterns and isoelectric point between CO₂-treated and non-treated control. The pH of CO₂-treated PPO stock solution was brought back from 5.4 to 8.0 after 6 weeks of frozen-storage and no enzyme reactivation was observed.     

Comparison of the inactivation kinetics of pectin methylesterases from carrot and peach by high-pressure carbon dioxide

The inactivation of pectin methylesterases (PMEs) from carrot and peach in buffer by high-pressure carbon dioxide (HPCD) at 55 °C was investigated. The two PMEs were effectively inactivated by HPCD, their residual activity (RA) decreasing with increasing pressures. The RA of the two PMEs exhibited a fast decrease firstly and reached a constant after a prolonged treatment time; their inactivation kinetics was adequately modelled by a fractional-conversion model. The non-zero RA(A_∞)of the two PMEs was 6–7%, with increasing pressures the kinetic rate constant, k, increased and the decimal reduction time, D, decreased for the HPCD-labile fraction of the two PMEs. The labile fraction of carrot PME was more susceptible to HPCD than that of peach PME; the activation volume, V_a, and Z_P (the temperature increase needed for a 90% reduction of D) was −1079.37 cm³/mol and 5.80 MPa for carrot PME, and −130.51 cm³/mol and 48.31 MPa for peach PME.