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<p>The present study was carried out to enhance the production of prot CT2 collagenolytic protease <i>by Bacillus halotolerans</i> CT2 isolate through the application of Plackett-Burman, Box-Behnken and Response surface methodology statistical designs. MgSO<sub>4</sub>7H<sub>2</sub>O, bean flour and incubation temperature were identified as significant factors affecting protease production using Plackett-Burman design. Box-Behnken and response surface methodology were also assessed and optimal experimental conditions were determined as follows: MgSO<sub>4</sub>.7H<sub>2</sub>O at 0.1&#xa0;g/L, bean flour at 0.5% and incubation temperature of 30&#xa0;°C. Under these optimized conditions, the predicted maximum collagenase activity reached 308.521 U/mL, representing a 16-fold increase compared to non-optimized conditions (19 U/mL). The application of 5 U of prot CT2 collagenolytic protease to beef meat for 2&#xa0;h at 40&#xa0;°C resulted in 51% reduction in meat shear force and 62% improvement in springiness, while preserving both the fresh color and moisture content of the meat. Scanning electron microscopy revealed substantial alterations in the muscle fiber organization following the protease treatment. Overall, our findings point out that prot CT2 collagenolytic protease, produced by <i>Bacillus halotolerans</i> under optimized conditions, show significant potential for utilization as an effective beef meat tenderizer.</p>

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Process improvement of prot CT2 collagenolytic protease production by Bacillus halotolerans with its application in beef tenderization

  • 期刊论文

  • 发表日期：

    2025年11月29日

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  • Dorra Gharbi,
  • Kais Djebali,
  • Ines Karkouch,
  • Moncef Chouaibi,
  • Ezzedine Aouani,
  • Ferid Limam,
  • Olfa Tabbene

摘要

The present study was carried out to enhance the production of prot CT2 collagenolytic protease by Bacillus halotolerans CT2 isolate through the application of Plackett-Burman, Box-Behnken and Response surface methodology statistical designs. MgSO47H2O, bean flour and incubation temperature were identified as significant factors affecting protease production using Plackett-Burman design. Box-Behnken and response surface methodology were also assessed and optimal experimental conditions were determined as follows: MgSO4.7H2O at 0.1 g/L, bean flour at 0.5% and incubation temperature of 30 °C. Under these optimized conditions, the predicted maximum collagenase activity reached 308.521 U/mL, representing a 16-fold increase compared to non-optimized conditions (19 U/mL). The application of 5 U of prot CT2 collagenolytic protease to beef meat for 2 h at 40 °C resulted in 51% reduction in meat shear force and 62% improvement in springiness, while preserving both the fresh color and moisture content of the meat. Scanning electron microscopy revealed substantial alterations in the muscle fiber organization following the protease treatment. Overall, our findings point out that prot CT2 collagenolytic protease, produced by Bacillus halotolerans under optimized conditions, show significant potential for utilization as an effective beef meat tenderizer.