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<p>Civet coffee is a premium specialty coffee known for its unique flavor, traditionally produced via spontaneous fermentation in the digestive system of civet cats (Paradoxurus hermaphrodites). This study aimed to replicate and optimize the fermentation process in vitro using eight bacterial strains isolated from civet gut, identified as Bacillus, Enterobacter, and Acinetobacter via 16&#xa0;S rRNA sequencing. Fermentation conditions were optimized using Response Surface Methodology (RSM) with a Box-Behnken Design, assessing the effects of temperature (25–35&#xa0;°C), time (24–48&#xa0;h), and inoculum concentration (5–15%) at 120&#xa0;rpm shaking. Coffee beans were fermented in flasks, dried to 10–12% moisture, roasted to Agtron 65 (medium), and evaluated by certified cuppers. Optimal conditions (30&#xa0;°C, 36&#xa0;h, and 10% inoculum) achieved a high cup score of 86.167, with final pH between 4.00 and 4.50. This correlated with enhanced flavor brightness, reduced bitterness, and microbial safety. Scaling up to a 5-liter bioreactor confirmed reproducibility. GC-MS analysis of inoculated beans showed elevated levels of key volatiles (aldehydes, esters, furans, and pyrazines), contributing to the coffee’s distinctive musky, nutty, and fruity aroma. A reduced caffeine content was also observed. These results present a controlled, scalable method for producing high-quality civet-like coffee through targeted microbial fermentation.</p>

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Gut-driven flavor: optimizing coffee fermentation using bacteria from civet cats

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • Enyesh Endalamaw,
  • Selamawit Tesfaye,
  • Mesfin Tafesse Gemeda

摘要

Civet coffee is a premium specialty coffee known for its unique flavor, traditionally produced via spontaneous fermentation in the digestive system of civet cats (Paradoxurus hermaphrodites). This study aimed to replicate and optimize the fermentation process in vitro using eight bacterial strains isolated from civet gut, identified as Bacillus, Enterobacter, and Acinetobacter via 16 S rRNA sequencing. Fermentation conditions were optimized using Response Surface Methodology (RSM) with a Box-Behnken Design, assessing the effects of temperature (25–35 °C), time (24–48 h), and inoculum concentration (5–15%) at 120 rpm shaking. Coffee beans were fermented in flasks, dried to 10–12% moisture, roasted to Agtron 65 (medium), and evaluated by certified cuppers. Optimal conditions (30 °C, 36 h, and 10% inoculum) achieved a high cup score of 86.167, with final pH between 4.00 and 4.50. This correlated with enhanced flavor brightness, reduced bitterness, and microbial safety. Scaling up to a 5-liter bioreactor confirmed reproducibility. GC-MS analysis of inoculated beans showed elevated levels of key volatiles (aldehydes, esters, furans, and pyrazines), contributing to the coffee’s distinctive musky, nutty, and fruity aroma. A reduced caffeine content was also observed. These results present a controlled, scalable method for producing high-quality civet-like coffee through targeted microbial fermentation.