ARTP法改良变色栓菌ATCC20869发酵漆酶

ARTP method to improve the fermentation laccase of Trametes versicolor ATCC20869

变色栓菌漆酶是一种含铜的多酚氧化酶,可氧化多种酚类和芳香族化合物,主产物为水且几乎不产生有毒副产物,具有较好的应用前景,但变色栓菌漆酶较低的产量限制了其进一步应用。本研究利用ARTP技术成功地从变色栓菌ATCC20869中突变出新的突变菌株TA-03。为了进一步提高漆酶产量,利用单因素摇瓶发酵实验和响应面分析法,优化了漆酶发酵条件。结果表明:在法尼醇添加量为4.48 mmol/L、硫酸铜添加量为1.90 mmol/L,pH为4.00的条件下,突变株漆酶产量达到了(283.77±8.91)U/mL。为深入探究突变菌株TA-03漆酶酶活性提高的原因,对其进行重测序和转录组分析。研究结果显示,漆酶活性区域氨基酸的突变以及新陈代谢等方面的富集导致了漆酶活性的提升。这为变色栓菌漆酶的实际应用进行了有意义的探索,提升了漆酶在发酵工业上的应用。

Trametes versicolor laccase is a copper-containing polyphenol oxidase that can oxidize a variety of phenolic and aromatic compounds.Its main product is water,and it produces almost no toxic by-products.While it has good application prospects,the low yield of Trametes versicolor laccase limits its further application.This study used Atmospheric and Room Temperature Plasma(ARTP)technology to successfully mutate a new strain,TA-03,from Trametes versicolor ATCC20869.To enhance laccase production,the paper optimized fermentation conditions through single-factor shake flask experiments and response surface analysis.The results showed that under optimized conditions,with a farnesol addition of 4.48 mmol/L,copper sulfate addition of 1.90 mmol/L,and pH maintained at 4.00,the laccase production of the mutant strain reached(283.77±8.91)U/mL.To explore the reasons for the increased laccase activity of the mutant strain TA-03,resequencing and transcriptome analysis were performed.The research results show that the mutation of amino acids in the active region of laccase and the enrichment of metabolism and other aspects lead to the improvement of laccase activity.This provides a meaningful exploration of the practical application of Trametes versicolor laccase and enhances the application of laccase in the fermentation industry.