Isanapong Jantiya, Suwannoi Kittikarn, Lertlattanapong Surangkana, Panchal Shweta
Faculty of Applied Science, Department of Agro-Industrial, Food and Environmental Technology, King Mongkut's University of Technology North Bangkok, 1518 Pracharat 1, Wongsawang, Bangsue, Bangkok, 10800 Thailand.
School of BioSciences and Technology, Vellore Institute of Technology, Vellore, 632014 India.
3 Biotech. 2024 Mar;14(3):73. doi: 10.1007/s13205-024-03926-7. Epub 2024 Feb 14.
This study is the first report on purification, characterization, and application of laccase derived from the white-rot fungus, HK35 (Hungary strain), in Congo Red decolorization. The purification process involved ammonium sulfate precipitation, dialysis, anion exchange chromatography, and ultrafiltration, yielding a specific laccase activity of 15.26 U/mg and a 30.21% recovery rate. The purified enzyme, with a molecular weight of approximately 34 kilodaltons, displayed optimal activity at a temperature of 60 °C and pH 4.0 when using 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) as a substrate. The enzyme maintained over 82.02 ± 1.01% of its activity at temperatures up to 50 °C after 180 min but displayed less than 5% of its activity at 60 and 70 °C. Notably, the enzyme's activity was significantly enhanced by Pb(NO), whereas β-mercaptoethanol completely inhibited the activity. Utilizing the Box-Behnken design, we optimized Congo Red decolorization efficiency to 91.05 ± 0.82% at 100 mg/L Congo Red, 1.5 mM mediator concentration, and 1.6 U/mL laccase activity. Analysis of Variance (ANOVA) suggested the model was significant, and all variables significantly influenced decolorization efficiency.
The online version contains supplementary material available at 10.1007/s13205-024-03926-7.
本研究首次报道了从白腐真菌HK35（匈牙利菌株）中提取的漆酶在刚果红脱色中的纯化、表征及应用。纯化过程包括硫酸铵沉淀、透析、阴离子交换色谱和超滤，得到的比漆酶活性为15.26 U/mg，回收率为30.21%。纯化后的酶分子量约为34千道尔顿，以2,2'-联氮双（3-乙基苯并噻唑啉-6-磺酸）（ABTS）为底物时，在60°C和pH 4.0的条件下表现出最佳活性。该酶在50°C下180分钟后仍保持其活性的82.02±1.01%以上，但在60°C和70°C下活性低于5%。值得注意的是，Pb(NO)可显著提高该酶的活性，而β-巯基乙醇则完全抑制其活性。利用Box-Behnken设计，在刚果红浓度为100 mg/L、介体浓度为1.5 mM、漆酶活性为1.6 U/mL的条件下，将刚果红脱色效率优化至91.05±0.82%。方差分析（ANOVA）表明该模型具有显著性，所有变量均对脱色效率有显著影响。
在线版本包含可在10.1007/s13205-024-03926-7获取的补充材料。
