pH Affects the Thermal Inactivation Parameters of R-Phycoerythrin from Porphyra yezoensis

ABSTRACT: R-phycoerythrin (PE), a protein that fluoresces in the visible range, was purified from Porphyra yezoensis. PE had a molecular mass of 292 kDa and an isoelectric point of 4.1 to 4.2. Thermal inactivation parameters of PE, calculated on the basis of fluorescence loss, were determined under different pH conditions. PE was more thermostable between pH 5.0 and 8.0, and became more heat sensitive at pH 4.0 and 10.0. PE at pH 6.0 had the highest D value (12258.7 min) at 70 °C. The z values of PE increased from 4.58 °C at pH 5.0 to 9.15 °C at pH 9.0. PE could be used as a time-temperature integrator by adjusting the inactivation kinetics of PE to match those of target microorganisms in a thermal process.     

Thermal Inactivation of Microorganisms

This paper serves as an overview of various aspects of thermal processing. Heat processing of foods has a long history and is still one of the most important preservation methods. To guarantee microbiological safety and stability, large safety margins are often applied in traditional heat processes. Because of the need for more fresh like foods, there is a need for milder preservation methods without compromising on safety and stability. The review deals with heat resistance data and mathematical models that describe heat inactivation. The effects of food composition are not yet fully clear and more knowledge of the cell physiology of the target microorganism could be of help in predicting the effects of food constituents. Finally, special attention has been paid to biological time temperature indicators to enable proper process calculations.     

The Deglet-Noor Date Polyphenoloxidase Activity Is Very Thermolabile and Exhibits Peculiarities in Its Thermal Inactivation Kinetics

In our current research on the preservation of the Deglet-Noor date quality during thermal processing, the heat inactivation of the polyphenoloxidase in enzyme extracts of this fruit was investigated. In contrast to previous studies on this enzyme in this exotic fruit, we performed full kinetic analyses at different temperatures (25-70°C) and enzyme activity was measured by the initial velocity methodology. The heat-inactivation of the polyphenoloxidase from the Deglet-Noor date does not obey a simple “one step-two stages” mechanism. All the recorded kinetics were described using a simple three parameters-biexponential model that we previously showed to be a common arranged form of kinetic expression for the known wide diversity of mechanisms of enzyme deactivation. The values of the two rate constants of the model were collected for four temperatures, ranging from 25 to 70°C and their variation was analyzed through the ARRHENIUS's law to determine the Ea values (11.83 and 54.71 kJ mol^-1, respectively). Possible explanations and implications of such low values are discussed. These results are compared to other results from laboratory as well as elsewhere in the literature.     

不同介质中甲型副伤寒沙门氏菌的热失活特性

研究甲型副伤寒沙门氏菌在不同介质中热失活规律,对接种10~8 CFU/g甲型副伤寒沙门氏菌的不同介质进行55、63、72℃热处理,测定处理后样品中甲型副伤寒沙门氏菌的菌体浓度。应用Dose Resp模型拟合在3种温度下志贺氏菌的动力学模型,用D值(decimal reduction time)表示甲型副伤寒沙门氏菌在不同介质中的耐热情况。结果表明:甲型副伤寒沙门氏菌的热失活曲线运用Dose Resp模型拟合较好,相关系数均在0.97以上。甲型副伤寒沙门氏菌在不同介质中的D值不同,在蛋白质和脂肪含量较高的灌肠肥瘦肉1∶1(m/m)中D值较大,相对耐热。55℃条件下热处理25.0 min,63℃热处理2.03 min,72℃热处理0.31 min后,一般肉制品中甲型副伤寒沙门氏菌均能被杀死。     

Effects of High‐Hydrostatic Pressure on Inactivation of Human Norovirus and Physical and Sensory Characteristics of Oysters

The purpose of the study was to determine the effect of high‐hydrostatic pressure (HHP) on inactivation of human norovirus (HuNoV) in oysters and to evaluate organoleptic characteristics of oysters treated at pressure levels required for HuNoV inactivation. Genogroup I.1 (GI.1) or Genogroup II.4 (GII.4) HuNoV was inoculated into oysters and treated at 300 to 600 MPa at 25 and 0 °C for 2 min. After HHP, viral particles were extracted by porcine gastric mucin‐conjugated magnetic beads (PGM‐MBs) and viral RNA was quantified by real‐time RT‐PCR. Lower initial temperature (0 °C) significantly enhanced HHP inactivation of HuNoV compared to ambient temperature (25 °C; P < 0.05). HHP at 350 and 500 MPa at 0 °C could achieve more than 4 log₁₀ reduction of GII.4 and GI.1 HuNoV in oysters, respectively. HHP treatments did not significantly change color or texture of oyster tissue. A 1‐ to 5‐scale hedonic sensory evaluation on appearance, aroma, color, and overall acceptability showed that pressure‐treated oysters received significantly higher quality scores than the untreated control (P < 0.05). Elevated pressure levels at 450 and 500 MPa did not significantly affect scores compared to 300 MPa at 0 °C, indicating increasing pressure level did not affect sensory acceptability of oysters. Oysters treated at 0 °C had slightly lower acceptability than the group treated at room temperature on day 1 (P < 0.05), but after 1 wk storage, no significant difference in sensory attributes and consumer desirability was observed (P > 0.05).     

熏煮火腿中志贺氏菌的热失活特性

为了研究志贺氏菌不同菌株在熏煮火腿中热失活规律,以55、63℃对接种108 CFU/g志贺氏菌的熏煮火腿进行热处理,测定处理后样品中志贺氏菌的残存菌体浓度。应用Logistic模型和3rd degree polynomial模型拟合在两种温度下志贺氏菌的动力学模型,用D值(decimal reduction time)表示志贺氏菌在熏煮火腿中的耐热情况。结果表明:11株志贺氏菌的热失活曲线运用3rd degree polynomial拟合较好,相关系数均在0.98以上。志贺氏菌不同菌株的D值不同,55℃条件下志贺氏菌在熏煮火腿中的D值为4.46~10.66 min,63℃条件下D值为0.48~3.22 min。不同菌株之间抗热性存在差异,其中菌株S-5的耐热性最强,S-6最弱。     

鲜枣脂氧合酶酶学特性及热失活动力学研究

以亚油酸为底物,采用紫外分光光度法测定鲜枣脂氧合酶(lipoxygenase,LOX)的酶学特性及热失活动力学性质。考察了温度、p H、金属离子及络合物对LOX活性的影响,并建立了LOX酶促反应动力学及热失活动力学参数。结果表明:鲜枣LOX的最大吸收波长为265 nm;最适温度和p H分别为40℃、7.0;在p H值为5.0~6.0的环境中,LOX活性保持相对稳定;酶促反应动力学符合米氏方程所描述的单底物酶促反应动力学,其Km和Vmax分别为0.14 mmol/L、0.26 U/min,鲜枣LOX与底物亚油酸的亲和力较好;且LOX的热失活遵循一级反应动力学规律,反应活化能为105.20 k J/mol;2 mmol/L Cu2+和10 mmol/L Cu2+、Ca2+、Mn2+对LOX酶活力有激活作用(P0.05),而2 mmol/L Mn2+、Zn2+、Mg2+和10 mmol/L Mg2+,EDTA对LOX活性具有抑制作用(P0.05)。该试验为鲜枣加工贮藏中LOX活性控制提供了有益的数据参考。     

Heat Inactivation Kinetics of Apple Polyphenoloxidase and Activation of its Latent Form

Heat inactivation kinetics of crude polyphenoloxidase (PPO) from six apple cultivars (Golden Delicious, Starking Delicious, Granny Smith; Gloster, Starcrimson and Amasya) were studied at three temperatures (68°, 73° and 78°C). PPO activity initially increased and then decreased with heat, following a first order kinetic model. Increase in activity indicated presence of latent PPO. Regression coefficients for the linear portions of inactivation curves were computed to determine inactivation parameters. Reaction data at 78°C revealed that PPO in Amasya was the least and Starking Delicious the most heat-stable. Rate constants for heat inactivation at 78°C ranged from 15.99–28.27. 10^?2 min^?1. Activation energies varied between 54.7–77.2 kcal. mol^?1 with z values of 7.1–10.0C°.PPO in apples was generally more heat-stable than PPO in most fruits.     

Inactivation Kinetics of Enzymes by Using Continuous Treatment with Microbubbles of Supercritical Carbon Dioxide

ABSTRACT Enzyme inactivation using a new apparatus for continuous treatment with microbubbles supercritical carbon dioxide (SC-CO₂) was investigated. D value of a-amylase (5.0±1.2 min) subjected to microbubbles of SC-CO₂ treatment (microbubbles-SCT) at 35 °C, 30 MPa was lower than that (227 ± 15.9 min) subjected to heat treatment (HT) at 70 °C. D value of acid protease was reduced by microbubbles-SCT at 50 °C, 30 MPa (15.4 ± 4.1 min), compared to HT at 50 °C (233 ± 15.2 min). The activation energy for the inactivation of acid protease (135 ± 8.3 kJ mol^-1) by microbubbles-SCT was 1 half of that (259 ± 9.0 kJ mol^-1) by HT. These results indicated that continuous treatment with microbubbles of SC-CO₂ was effective for enzyme inactivation.     

Effect of desiccation, starvation, heat, and cold stresses on the thermal resistance of Enterobacter sakazakii in rehydrated infant milk formula

Enterobacter sakazakii is an opportunistic foodborne pathogen that has been isolated from powdered infant milk formula. This study determined the effect of desiccation, starvation, heat and cold stresses on the thermal inactivation of E. sakazakii in rehydrated infant milk formula (RIMF). Stressed cells were mixed with RIMF at 52, 54, 56, and 58 degrees C for various time periods. The D- and z-values were determined by using linear regression analysis. D-values for unstressed E. sakazakii at 52, 54, 56, and 58 degrees C were 15.33, 4.53, 2, and 0.53 min, respectively. Desiccation and heat stresses, but not starvation or cold stress, caused significant (P < 0.05) reduction in D-values. The z-values of desiccated, starved, heat stressed, and cold stressed E. sakazakii were not significantly different from unstressed cells (4.22 degrees C). Thermal resistance of E. sakazakii in RIMF is affected by the environmental stresses; that is, desiccation and heat stresses that may surround the bacterium prior to the contamination of infant formula. The results of this study may be of use to regulatory agencies, infant milk producers, and infant caregivers to design heating processes to eliminate E. sakazakii that may be present in infant milk formula.     

Thermal Inactivation Kinetics of Peroxidase and Lipoxygenase from Broccoli, Green Asparagus and Carrots

ABSTRACT: Thewermal inactivation curves for peroxidase (POD) and lipoxygenase (LOX) in broccoli (florets), green asparagus (tip and stem), and carrots (cortex and core) extracts were determined in the range of 70 to 95 °C for 0 to 600 s. The capillary tube method was used to obtain quasi-isothermal conditions. The kinetics of both enzymes showed a biphasic first-order model, while at 70 °C, LOX in asparagus showed a monophasic first-order behavior. LOX activity was not detected for carrots. Kinetic parameters, k and Ea , were determined for heat-labile and heatresistant isoenzyme fractions. Additionally, initial and residual activities for both enzymes within tissue sections showed a different distribution and heat stability.     

Comparative Study of Thermal Inactivation of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in Ground Pork

ABSTRACT: Thermal inactivation of Escherichia coli O157:H7, Salmonella , and Listeria monocytogenes in ground pork was compared. The D (decimal reduction time at a certain heating temperature) values of E. coli O157:H7, Salmonella , and L. monocytogenes at 55 to 70°C were 33.44 to 0.048 min, 45.87 to 0.083 min, and 47.17 to 0.085 min, respectively. The z (temperature rise for 1 log10 reduction of D) value of E. coli O157:H7, Salmonella , and L. monocytogenes in ground pork was 4.94°C, 5.89°C, and 5.92°C, respectively. Significant difference was found on the D and z values between E. coli O157:H7 and Salmonella or between E. coli O157:H7 and L. monocytogenes . The D and z values of Salmonella in ground pork were not significantly different from L. monocytogenes .     

Thermal inactivation of hepatitis A virus in suspension and in dried mussels (Mytilus edulis)

We examined the effects of three different temperatures (60, 85 and 100 °C) and durations of time (1 min at 100  ° C to 30 min at 60  ° C) on hepatitis A virus (HAV) in suspension and dried mussels (Mytilus edulis). In suspension, 3.61, 4.48, 5.06 and 5.66 log₁₀ tissue culture infectious dose (TCID)₅₀/mL were reduced by 60  ° C for 5 min, 60  ° C for 15 min, 60  ° C for 30 min and 85  ° C for 3 min, respectively. In dried mussels, 1.34, 1.94, 3.16, 2.36, 3.53 and 4.38 log₁₀TCID₅₀/mL were reduced by 60  ° C for 5 min, 60  ° C for 15 min, 60  ° C for 30 min, 85  ° C for 3 min, 85  ° C for 6 min and 85  ° C for 10 min, respectively. HAV inactivation from suspension and dried mussels was achieved by 85  ° C for 6 min and 15 min, respectively, and also by a 1 min at 100  ° C. At 60, 85 and 100  ° C, the 1‐log (D‐values) inactivation from both suspension and dried mussels was 6.33 and 7.93, 0.98 and 3.05, and 0.28 and 0.38 min, respectively. A higher temperature and/or a thermal treatment time shorter than 85  ° C for 6 min (100  ° C for 1 min) could be used for commercial target foods for complete HAV inactivation.     

Efficacy of Combined Sous Vide‐Microwave Cooking for Foodborne Pathogen Inactivation in Ready‐to‐Eat Chicory Stems

There is a variety of different food processing methods, which can be used to prepare ready‐to‐eat foods. However, the need to preserve the freshness and nutritional qualities leads to the application of mild technologies which may be insufficient to inactivate microbial pathogens. In this work, fresh chicory stems were packed under a vacuum in films, which were transparent to microwaves. These were then exposed to microwaves for different periods of time. The application of sous vide microwave cooking (SV‐MW, 900 W, 2450 MHz), controlled naturally occurring mesophilic aerobic bacteria, yeasts and molds for up to 30 d when vacuum‐packed vegetables were stored at 4 °C. In addition, the process lethality of the SV‐MW 90 s cooking was experimentally validated. This treatment led to 6.07 ± 0.7 and 4.92 ± 0.65 log cfu/g reduction of Escherichia coli and Listeria monocytogenes inoculated over the chicory stems (100 g), respectively. With an initial load of 9 log cfu/g for both pathogens, less than 10 cfu/g of surviving cells were found after 90 s cooking. This shows that short‐time microwave cooking can be used to effectively pasteurize vacuum‐packed chicory stems, achieving >5 log cfu/g reduction of E. coli and L. monocytogenes.     

Determination of Trimethylamine in Spinach,Cabbage, and Lettuce at Alkaline pH by Headspace Solid‐Phase Microextraction

Abstract: Trimethylamine (TMA) found in some leafy vegetables, such as spinach, cabbage, and lettuce, at alkaline pH was identified and quantified using headspace solid‐phase microextraction and gas chromatography‐mass spectrometry (HS‐SPME and GC‐MS). HS‐SPME conditions were optimized at an adsorption temperature of 50 °C, equilibration time of 5 min, and adsorption time of 5 min with 65 μm of polydimethylsiloxane/divinylbenzene fiber. The TMA that was formed from spinach, cabbage, and lettuce was assayed at pH 7 to 11 for 0 to 4 h at 50 °C using HS‐SPME. The results showed that the amount of TMA formed was dependent on pH. The amount of TMA formed increased dramatically at a pH greater than 9. TMA was not formed at a pH lower than 7. Spinach produced a higher amount of TMA than cabbage or lettuce. TMA was formed at alkaline pH from choline, betaine, and carnitine, which are TMA precursors. To confirm the SPME results, TMA was quantitated using the AOAC official method. Data obtained from chemical analysis were in good agreement with the SPME data. The formation mechanism of TMA is thought to be the Hofmann elimination reaction, which generates amine compounds at alkaline pH. Practical Application: Fishy off‐flavor in foods is associated with trimethylamine (TMA), which is frequently found in fish and seafood. In this study, TMA was identified for the first time in some leafy vegetables, such as spinach, cabbage, and lettuce, at alkaline pH. The presence of TMA in leafy vegetables under certain circumstances such as high pH and temperature may affect the sensory properties of foods containing these vegetables.     

Kinetics of Thermal Inactivation of Peroxidase and Color Degradation of African Cowpea (Vigna unguiculata) Leaves

Cowpea leaves form an important part of the diet for many Kenyans, and they are normally consumed after a lengthy cooking process leading to the inactivation of peroxidase (POD) that could be used as an indicator for the potential shelf life of the vegetables. However, color degradation can simultaneously occur, leading to poor consumer acceptance of the product. The kinetics of POD in situ thermal (for thermal treatments in the range of 75 to 100 °C/120 min) inactivation showed a biphasic first‐order model, with Arrhenius temperature dependence of the rate constant. The kinetic parameters using a reference temperature (T_ref) of 80 °C were determined for both the heat‐labile phase (k_ref = 11.52 ± 0.95 × 10^?2 min^?1 and E_a of 109.67 ± 6.20 kJ/mol) and the heat‐stable isoenzyme fraction (k_ref = 0.29 ± 0.07 × 10^?2 min^?1 and E_a of 256.93 ± 15.27 kJ/mol). Color degradation (L*, a*, and b* value) during thermal treatment was investigated, in particular as the “a*” value (the value of green color). Thermal degradation (thermal treatments between 55 and 80 °C per 90 min) of the green color of the leaves followed a fractional conversion model and the temperature dependence of the inactivation rate constant can be described using the Arrhenius law. The kinetic parameters using a reference temperature (T_refC = 70 °C) were determined as k_refC = 13.53 ± 0.01 × 10^?2 min^?1 and E_aC = 88.78 ± 3.21 kJ/mol. The results indicate that severe inactivation of POD (as an indicator for improved shelf life of the cooked vegetables) is accompanied by severe color degradation and that conventional cooking methods (typically 10 min/100 °C) lead to a high residual POD activity suggesting a limited shelf life of the cooked vegetables.     

Thermal Inactivation Kinetics of Salmonella and Listeria in Ground Chicken Breast Meat and Liquid Medium

ABSTRACT: Thermal inactivation of Listeria innocua and 6 Salmonella serotypes in ground chicken breast meat was compared to that in peptone (0.1%) - agar (0.1%) solution. Inoculated samples were packed in a thin-wall metal tube and submerged in a water bath at temperatures ranging from 55.0 to 70.0 °C. For Salmonella and Listeria , the D values in ground chicken breast meat at 55 to 70 °C were higher (p < 0.0001) than those in peptone-agar solution; however, the z values were not significantly different. Complete first-order inactivation models, with Arrhenius temperature dependency, were developed for each inoculum and medium.     

A Gompertz Model Approach to Microbial Inactivation Kinetics by High‐Pressure Processing (HPP): Model Selection and Experimental Validation

A recently proposed Gompertz model (GMPZ) approach describing microbial inactivation kinetics by high‐pressure processing (HPP) incorporated the initial microbial load (N₀) and lower microbial quantification limit (N_lim), and simplified the dynamic effects of come‐up time (CUT). The inactivation of Listeria innocua in milk by HPP treatments at 300, 400, 500, and 600 MPa and pressure holding times (t_hold) ≤10 min was determined experimentally to validate this model approach. Models based on exponential, logistic‐exponential, and inverse functions were evaluated to describe the effect of pressure on the lag time (λ) and maximum inactivation rate (μ_max), whereas the asymptote difference (A) was fixed as A = log₁₀(N₀/N_lim). Model performance was statistically evaluated and further validated with additional data obtained at 450 and 550 MPa. All GMPZ models adequately fitted L. innocua data according to the coefficient of determination (R² ≥ 0.95) but those including a logistic‐exponential function for μ_max(P) were superior (R² ≥ 0.97). These GMPZ versions predicted that approximately 597 MPa is the theoretical pressure level (P_λ) at which microbial inactivation begins during CUT, mathematically defined as λ (P = P_λ) = t_CUT, and matching the value observed on the microbial survival curve at 600 MPa. As pressure increased, predictions tended to slightly underestimate the HPP lethality in the tail section of the survival curve. This may be overseen in practice since the observed microbial counts were below the predicted log₁₀ N values. Overall, the modeling approach is promising, justifying further validation work for other microorganisms and food systems.     

Predictive Modeling of the Effect of ε‐Polylysine Hydrochloride on Growth and Thermal Inactivation of Listeria monocytogenes in Fish Balls

This study aimed to evaluate the effect of ε‐polylysine hydrochloride (ε‐PLH) on the growth and thermal inactivation kinetics of Listeria monocytogenes in fish balls. Samples, supplemented with ε‐PLH (0, 150, or 300 ppm, w/w), were inoculated with a three‐strain cocktail of L. monocytogenes and incubated at constant temperatures of 3.4, 8, 12, or 16 °C for growth studies, or heated at 60, 62.5, 65, or 67.5 °C for thermal inactivation tests. The growth curves were fitted to the Huang primary model, and the Huang and Ratkowsky square‐root models (SRM) were used as the secondary models to evaluate the effect of temperature on bacterial growth. The survival during heating was analyzed with a log‐linear model. The results showed that, while the lag time of L. monocytogenes was affected by both ε‐PLH concentration and temperature, the specific growth rate was unaffected by ε‐PLH. Under the same temperature, a 10‐time in increase of the lag time would be expected for every 565 ppm in the increase of ε‐PLH concentration. Using the Ratkowsky SRM, the estimated nominal minimum growth temperature was –2.04 °C, while the minimum growth temperature was 0.29 °C when estimated with the Huang SRM. Validation at 10 °C showed that the Huang primary model, in combination with either the Huang or Ratkowsky SRM, could accurately predict the growth of L. monocytogenes. On the other hand, the thermal resistance of the pathogen was significantly reduced by increase in temperature or ε‐PLH. The thermal z value of L. monocytogenes was 5.78 °C, and the ε‐PLH z value was 1642 ppm. The results of this study showed that the combined application of ε‐PLH and temperature can be used to control L. monocytogenes in fish balls and to improve food safety and reduce risks to public health.