Ma Yaping, Liu Minchang, Gu Ruifang, Zhang Rongya, Ji Xiaomei, Zhang Juan, Wen Wu, Peng Zheng
Technology Center, China Tobacco Sichuan Industrial Co., Ltd., Chengdu, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
Front Bioeng Biotechnol. 2025 Aug 15;13:1631687. doi: 10.3389/fbioe.2025.1631687. eCollection 2025.
Laccase exhibits significant applications in food additives, wastewater treatment, and biomass processing. Fungal laccase exhibits high activity, good stability, and excellent performance. However, scale-up production of high-yield laccase from fungi is challenging. This study aimed to identify crucial factors affecting enzyme production and analyze the enzymatic properties of laccase during fermentation.
A laccase-producing white-rot fungus was used for fermentation process optimization in 200 L and 1200 L fermenters. The Plackett-Burman design revealed three significant influencing factors: temperature, aeration ratio, and agitation speed. The steepest ascent experiment was used to approximate the maximum response region, followed by the establishment of a regression model between experimental factors and laccase activity using the Box-Behnken response surface methodology and optimal fermentation condition selection.
The optimal conditions for laccase production by fermentation were 30°C temperature, 0.66 aeration ratio, and 100 rpm agitation speed, achieving a maximum laccase activity of 214,185.2 U/L. Dissolved oxygen (DO) was a crucial factor for high laccase yield, and its maintenance at a high level in the fermentation system significantly enhanced the enzyme activity. Fermentation batches with varying laccase production levels exhibited a trend of decreasing pH and a subsequent increase in the mid-to-late fermentation stages. With decreased pH, the DO level started declining; when DO stabilized, the pH started to rebound, coinciding with the peak laccase activity, indicating a signal of fermentation endpoint in industrial production.
This study provides valuable theoretical and data support for the industrial production of laccase by fungi through optimized fermentation processes.
漆酶在食品添加剂、废水处理和生物质加工中具有重要应用。真菌漆酶具有高活性、良好的稳定性和优异的性能。然而，从真菌中大规模生产高产漆酶具有挑战性。本研究旨在确定影响酶生产的关键因素，并分析发酵过程中漆酶的酶学性质。
使用一种产漆酶的白腐真菌在200升和1200升发酵罐中优化发酵过程。Plackett-Burman设计揭示了三个显著影响因素：温度、通气比和搅拌速度。采用最速上升实验来逼近最大响应区域，随后使用Box-Behnken响应面方法建立实验因素与漆酶活性之间的回归模型，并选择最佳发酵条件。
发酵产漆酶的最佳条件为温度30°C、通气比0.66和搅拌速度100转/分钟，漆酶最大活性达到214,185.2 U/L。溶解氧（DO）是高产漆酶的关键因素，在发酵系统中保持其高水平可显著提高酶活性。不同漆酶生产水平的发酵批次在发酵中后期呈现pH值下降然后上升的趋势。随着pH值下降，DO水平开始下降；当DO稳定时，pH值开始反弹，与漆酶活性峰值同时出现，这表明在工业生产中是发酵终点的信号。
本研究通过优化发酵工艺为真菌工业生产漆酶提供了有价值的理论和数据支持。
