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<p>The high potential of the probiotic <i>L. reuteri</i> CRL 1098, as hypocholesterolemic and vitamin B<sub>12</sub> producer strain, would require addressing freeze-drying studies of the microorganism as well as its stability during storage. <i>L. reuteri</i> CRL 1098 was freeze-dried with glutamate, maltodextrin and trehalose as lyoprotectants and an accelerated inactivation assays under isothermal conditions (50, 60 and 70°C) was used to evaluate the stability of lyophilizates. Results obtained were analyzed with informatic GInaFiT Ap-inn and traditional predictive tools (D/z and Arrhenius models) for describing both, the inactivation process and the bacterial behaviour. The predicted shelf life of the lyophilized probiotic to obtain 1 × 10<sup>6</sup> UFC/g was experimentally tested at 30°C. Regardless of the lyoprotectant used, the survival of <i>L. reuteri</i> CRL1098 fitted acceptably to the weilbulian model of Marfat in all heat treatments; the data described curves with up dawn concavities (<i>p</i> &lt; 1) and allowed to identify a more sensitive bacterial population through the first decimal reduction time (<i>δ</i>-value) specially at 50°C in presence of maltodextrin. From the second reduction time, survivors of <i>L. reuteri</i> CRL 1098 fitted to first kinetic order reactions (lineal distribution of data), so <i>D</i>-values (decimal reduction time), <i>k</i>-values (inactivation rate constant), and z-values (intrinsic resistance) as well as the activation energy Ea, were calculated through D/z model and Arrhenius model. From these, a resistant subpopulation of <i>L. reuteri</i> CRL 1098 could be described, mainly when trehalose was used as lyoprotectant, which was characterized by high z-values. These results correlated with higher <i>D</i>-values but lower <i>k</i>-values and Ea in trehalose, which did not show the best stabilizing property along the storage tested at 30°C. Maltodextrine proved to be the best stabilizer lyoprotectant, especially when the shelf life is predicted at low temperatures of conservation.</p>

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Selection of lyoprotectants for the stabilization of the freeze-dried probiotic Limosilactobacillus reuteri CRL 1098 by an accelerated inactivation assay

  • 期刊论文

  • 发表日期：

    2025年9月16日

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  • María Inés Torino,
  • Graciela Font,
  • María José Fornaguera,
  • María Pía Taranto

摘要

The high potential of the probiotic L. reuteri CRL 1098, as hypocholesterolemic and vitamin B12 producer strain, would require addressing freeze-drying studies of the microorganism as well as its stability during storage. L. reuteri CRL 1098 was freeze-dried with glutamate, maltodextrin and trehalose as lyoprotectants and an accelerated inactivation assays under isothermal conditions (50, 60 and 70°C) was used to evaluate the stability of lyophilizates. Results obtained were analyzed with informatic GInaFiT Ap-inn and traditional predictive tools (D/z and Arrhenius models) for describing both, the inactivation process and the bacterial behaviour. The predicted shelf life of the lyophilized probiotic to obtain 1 × 106 UFC/g was experimentally tested at 30°C. Regardless of the lyoprotectant used, the survival of L. reuteri CRL1098 fitted acceptably to the weilbulian model of Marfat in all heat treatments; the data described curves with up dawn concavities (p < 1) and allowed to identify a more sensitive bacterial population through the first decimal reduction time (δ-value) specially at 50°C in presence of maltodextrin. From the second reduction time, survivors of L. reuteri CRL 1098 fitted to first kinetic order reactions (lineal distribution of data), so D-values (decimal reduction time), k-values (inactivation rate constant), and z-values (intrinsic resistance) as well as the activation energy Ea, were calculated through D/z model and Arrhenius model. From these, a resistant subpopulation of L. reuteri CRL 1098 could be described, mainly when trehalose was used as lyoprotectant, which was characterized by high z-values. These results correlated with higher D-values but lower k-values and Ea in trehalose, which did not show the best stabilizing property along the storage tested at 30°C. Maltodextrine proved to be the best stabilizer lyoprotectant, especially when the shelf life is predicted at low temperatures of conservation.