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<p>Feruloyl esterase (FAE) plays a crucial role in the biodegradation of agricultural waste. However, its limited enzymatic activity hinders its broader application. To maximize the FAE activity of the recombinant <i>Escherichia coli</i> strain pGEX-4T-1-BpFae<sup>T132C−D143C</sup>, we systematically optimized its fermentation conditions. Furthermore, we explored its potential application in the enzymatic hydrolysis of rice husk for the simultaneous production of ferulic acid (FA) and xylo-oligosaccharides (XOS). The optimal fermentation conditions were systematically determined through a combination of single-factor experiments, Plackett-Burman (PB) design, steepest ascent path design, and response surface methodology (RSM). Under these optimized conditions—initial pH of 7.0, lactose concentration of 4.95&#xa0;g/L, post-induction temperature of 25.8&#xa0;°C, shaker rotational speed of 200&#xa0;rpm, inoculum size of 0.1%, loading volume of 12.5 mL/250 mL, pre-induction time of 9.2&#xa0;h, and post-induction time of 36&#xa0;h in Terrific Broth medium—the activity of BpFae<sup>T132C−D143C</sup> reached 4.43 U/mL, representing a 2.91-fold increase compared to the single-factor experiment. In a 3&#xa0;L bioreactor, the total enzyme activity achieved was 1.35 U/mL. When BpFa<sup>eT132C−D143C</sup> was combined with xylanase McXyn0243 for the enzymatic hydrolysis of rice husk, the yields of FA and XOS reached 67.17% and 61.59%, respectively. This demonstrated a significant synergistic effect, outperforming the results obtained when either enzyme was used independently.</p>

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Optimization of lactose-induced expression conditions for feruloyl esterase BpFaeT132C−D143C and its application in simultaneous production of ferulic acid and xylo-oligosaccharides from rice husk

  • 期刊论文

  • 发表日期：

    2026年2月25日

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  • Zechen Wang,
  • Jinghao Ma,
  • Yanbin Tang,
  • Xiangqin Lou,
  • Rana Abdul Basit,
  • Zhilei Fu,
  • Jinpeng Wang,
  • Guangsen Fan

摘要

Feruloyl esterase (FAE) plays a crucial role in the biodegradation of agricultural waste. However, its limited enzymatic activity hinders its broader application. To maximize the FAE activity of the recombinant Escherichia coli strain pGEX-4T-1-BpFaeT132C−D143C, we systematically optimized its fermentation conditions. Furthermore, we explored its potential application in the enzymatic hydrolysis of rice husk for the simultaneous production of ferulic acid (FA) and xylo-oligosaccharides (XOS). The optimal fermentation conditions were systematically determined through a combination of single-factor experiments, Plackett-Burman (PB) design, steepest ascent path design, and response surface methodology (RSM). Under these optimized conditions—initial pH of 7.0, lactose concentration of 4.95 g/L, post-induction temperature of 25.8 °C, shaker rotational speed of 200 rpm, inoculum size of 0.1%, loading volume of 12.5 mL/250 mL, pre-induction time of 9.2 h, and post-induction time of 36 h in Terrific Broth medium—the activity of BpFaeT132C−D143C reached 4.43 U/mL, representing a 2.91-fold increase compared to the single-factor experiment. In a 3 L bioreactor, the total enzyme activity achieved was 1.35 U/mL. When BpFaeT132C−D143C was combined with xylanase McXyn0243 for the enzymatic hydrolysis of rice husk, the yields of FA and XOS reached 67.17% and 61.59%, respectively. This demonstrated a significant synergistic effect, outperforming the results obtained when either enzyme was used independently.