The purification and characterization of a novel alkali-stable pectate lyase produced by Bacillus subtilis PB1
The purification and characterization of a novel alkali-stable pectate lyase produced by Bacillus subtilis PB1
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DOI：
10.1007/s11274-017-2357-8
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发表时间：
2017-10
影响因子：
4.1
通讯作者：
Man Zhou;Jingli Wu;Tao Wang;Lina Gao;Huijun Yin;Xin Lü
中科院分区：
文献类型：
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作者：
Man Zhou;Jingli Wu;Tao Wang;Lina Gao;Huijun Yin;Xin Lü
果胶酶是一种重要的酶，具有广泛的工业用途，其中细菌生产的果胶酶与真菌生产的果胶酶相比是稀缺的。本研究首次从烤烟叶片中分离到一种产生细胞外果胶酶的新型细菌，并根据其16S rRNA基因鉴定为枯草芽孢杆菌pb1。经硫酸铵沉淀、离子交换、凝胶过滤层析纯化果胶溶酶，SDS-PAGE测定分子量为43.1±0.5 kDa。利用MALDI-TOF质谱对该果胶酶进行肽质量指纹图谱分析，结果表明该酶与果胶裂解酶具有同源性，酶标为BsPel-PB1。BsPel-PB1的最适温度为50℃。BsPel-PB1的最适pH为pH 9.5， pH稳定范围为5 ~ 11。kmandvmax分别为0.312 mg/mL和1248 U/mL。因此，BsPel-PB1是一种新型碱性果胶裂解酶，在果胶裂解相关行业具有潜在的商业应用前景。
Pectinase is an important kind of enzyme with many industrial applications, among which pectinases produced by bacteria were scarce compared with fungal sources. In this study, a novel bacterium which produced extracellular pectinase was firstly isolated from flue-cured tobacco leaves and identified asBacillus subtilisPB1 according to its 16S rRNA gene. The pectinolytic enzyme was purified by ammonium sulfate precipitation, ion-exchange and gel filtration chromatography, after which molecular weight was determined as 43.1 ± 0.5 kDa by SDS–PAGE. Peptide mass fingerprinting of the pectinase by MALDI-TOF MS showed that the purified enzyme shared homology with pectate lyase and was designated as BsPel-PB1. The optimal temperature for BsPel-PB1 was 50 °C. The optimal pH was pH 9.5 for BsPel-PB1 while it had a broad pH stability from 5 to 11. The values ofKmandVmaxwere 0.312 mg/mL and 1248 U/mL, respectively. Accordingly, the BsPel-PB1 was a novel alkaline pectate lyase which could find potential application as a commercial candidate in the pectinolytic related industries.