Isolation and Characterization of a Novel Laccase for Lignin Degradation, LacZ1

• Published in: Applied and environmental microbiology 2021-11, Vol.87 (23), p.e0135521-e0135521
• Main Authors: Zhang, Weiran, Wang, Weiwei, Wang, Jinghong, Shen, Guinan, Yuan, Yuan, Yan, Lei, Tang, Hongzhi, Wang, Weidong
• Format: Article
• Language: English
• Subjects: Affinity chromatography; Bacillus - enzymology; Bacillus - genetics; Benzoic acid; Biodegradation; Chelation; Chromatography; Copper; Degradation; Degradation products; Enzymatic activity; Enzyme activity; Enzymes; Gas chromatography; High temperature; Laccase; Laccase - genetics; Laccase - isolation & purification; Lignin; Lignin - metabolism; Lignocellulose; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Microorganisms; Molecular weight; Molecular weight distribution; NMR; Nuclear magnetic resonance; Phylogeny; Polymers; Protocatechuic acid; Reverse transcription
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.01355-21

Lignin is a complex natural organic polymer and is one of the primary components of lignocellulose. The efficient utilization of lignocellulose is limited because it is difficult to degrade lignin. In this study, we screened a gene fragment encoding laccase from the macrotranscriptome data of a microbial consortium WSC-6, which can efficiently degrade lignocellulose. The reverse transcription-quantitative PCR (RT-qPCR) results demonstrated that the expression level of the gene during the peak period of lignocellulose degradation by WSC-6 increased by 30.63 times compared to the initial degradation period. Phylogenetic tree analysis demonstrated that the complete gene is derived from a sp. and encoded laccase. The corresponding protein, LacZ1, was expressed and purified by Ni-chelating affinity chromatography. The optimum temperature was 75°C, the optimum pH was 4.5, and the highest enzyme activity reached 16.39 U/mg. We found that Cu was an important cofactor needed for LacZ1 to have enzyme activity. The molecular weight distribution of lignin was determined by gel permeation chromatography (GPC), and changes in the lignin structure were determined by H nuclear magnetic resonance ( H NMR) spectra. The degradation products of lignin by LacZ1 were determined by gas chromatography and mass spectrometry (GC-MS), and three lignin degradation pathways (the gentian acid pathway, benzoic acid pathway, and protocatechuic acid pathway) were proposed. This study provides insight into the degradation of lignin and new insights into high-temperature bacterial laccase. Lignin is a natural aromatic polymer that is not easily degraded, hindering the efficient use of lignocellulose-rich biomass resources, such as straw. Biodegradation is a method of decomposing lignin that has recently received increasing attention. In this study, we screened a gene encoding laccase from the lignocellulose-degrading microbial consortium WSC-6, purified the corresponding protein LacZ1, characterized the enzymatic properties of laccase LacZ1, and speculated that the degradation pathway of LacZ1 degrades lignin. This study identified a new, high-temperature bacterial laccase that can degrade lignin, providing insight into lignin degradation by this laccase.