The Determination of Assay for Laccase of Bacillus subtilis WPI with Two Classes of Chemical Compounds as Substrates
The Determination of Assay for Laccase of Bacillus subtilis WPI with Two Classes of Chemical Compounds as Substrates
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DOI：
10.1007/s12088-012-0298-3
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发表时间：
2012-08
影响因子：
3
通讯作者：
F. Sheikhi;M. Roayaei Ardakani;N. Enayatizamir;S. Rodríguez-Couto
中科院分区：
文献类型：
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作者：
F. Sheikhi;M. Roayaei Ardakani;N. Enayatizamir;S. Rodríguez-Couto
木质素分解酶复合体参与了木质素的降解。其中漆酶表现突出，因为它们使用分子氧作为共底物，而不是使用过氧化氢作为过氧化氢。芽孢杆菌属细菌漆酶最早是在Claus和Filip(Microbiol Res 152：209-216,1997)中报道的，此后又发现了更多的细菌漆酶。本研究从制浆造纸废水、山竹和甘蔗根际中筛选出漆酶产生菌。使用含有0.5 mm愈创木酚的营养琼脂培养基。结果表明，漆酶产生菌呈棕色，从中鉴定出16株芽孢杆菌。经生化试验和16S rDNA序列分析，鉴定其中一株为枯草芽孢杆菌WPI。该菌株对氧化底物ABTS和愈创木酚表现出类似漆酶的活性。本研究以愈创木酚为底物进行漆酶活力测定。为测定该菌株的漆酶活力，本研究首次采用愈创木酚为底物进行测定。SDS-PAGE和Native-PAGE证实了漆酶的存在。
Ligninolytic enzyme complexes are involved in lignin degradation. Among them laccases are outstanding because they use molecular oxygen as a co-substrate instead of hydrogen peroxide as used by peroxidases. Bacterial laccase of Bacillus genus was first reported in Claus and Filip (Microbiol Res 152:209–216, 1997), since then more bacterial laccases have been found. In this research, laccase-producing bacteria were screened from pulp and paper industry wastewater, bagass and sugarcane rhizosphere. Nutrient agar medium containing 0.5 mM of guaiacol was used. It was observed that the laccase-producing strains developed brown colour from which 16 strains ofBacilluswere identified. One of the isolated strains was identified asBacillus subtilisWPI based on the results of biochemical tests and 16S rDNA sequence analysis. This strain showed laccase-like activity towards the oxidizing substrates ABTS and guaiacol. In this study guaiacol was used as the substrate of laccase activity assay. For determination of laccase activity of this isolate guaiacol was used as a substrate of assay for the first time in this study. SDS-PAGE and Native-PAGE confirmed the presence of laccase.