Archives of microbiology: screening of pectinase-producing bacteria from citrus peel and characterization of a recombinant pectate lyase with applied potential
Jazyk angličtina Země Německo Médium print-electronic
Typ dokumentu časopisecké články
Grantová podpora
31701844
National Natural Science Foundation of China
2018J05057
Natural Science Foundation of Fujian province
JAT170078
Foundation of Fujian Educational Bureau
RVO: 60077344
Institutional support of the Biology Centre CAS, Institute of Entomology
2019T018
Fuzhou University Testing Fund of precious apparatus
PubMed
31932863
DOI
10.1007/s00203-020-01807-0
PII: 10.1007/s00203-020-01807-0
Knihovny.cz E-zdroje
- Klíčová slova
- Bacillus sp., Enzyme properties, Heterologous expression, Pectate lyase, PelB,
- MeSH
- Bacillus enzymologie genetika metabolismus MeSH
- Citrus metabolismus MeSH
- Escherichia coli genetika metabolismus MeSH
- kinetika MeSH
- koncentrace vodíkových iontů MeSH
- pektiny metabolismus MeSH
- polygalakturonasa biosyntéza metabolismus MeSH
- polysacharid-lyasy metabolismus MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- pectate lyase MeSH Prohlížeč
- pektiny MeSH
- polygalakturonasa MeSH
- polysacharid-lyasy MeSH
Pectinase is widely used in numerous industrial fields, including the food, wine, and paper industries. In this work, seven bacteria were isolated from orange peel and their pectinase production activity was assayed. One bacterium (OR-B2) identified as a Bacillus sp. showed the highest enzyme activity towards others. A gene encoding a pectate lyase designed as PelB-B2 in this work was amplified and heterogeneous expressed in E.coli. PelB-B2 was defined as a member of the PelB pectate lyase family after phylogenic tree analysis. 3D model of PelB-B2 was constructed by SWISS-MODEL and PelB-B2 showed conserved para-β structure. After inducing culture and purified by Ni-affinity chromatography, the properties of the purified PelB-B2 were assayed. Optimal pH and temperature for PelB-B2 was pH 8.0 and 50 °C, respectively. PelB-B2 showed excellent pH stability and thermostability. It was stable within pH range 3.0-11.0 and retained more than 51% activity after incubation at 40 °C, 50 °C, or 60 °C for 1 h. Furthermore, we determined that PelB-B2 was a Ca2+-dependent pectinase and the pectin extracted from citrus was the benefit substrate for PelB-B2. The Km and Vmax of PelB-B2 were 1.64 g/L and 232.56 mol/(L min), respectively. The OR-B2 can be a new resource for pectinase production and the PelB-B2 has potential for industrial application. 7 bacteria were isolated from orange peel, namely OR-B1 to OR-B7 and their pectinase activities were assayed. One pectate lyase belongs to PelB family was cloned from OR-B2 and heterogeneous expressed in E. coli. Purified PelB-B2 was further studied with its properties. Effects of pH, temperature, chemicals, substrate on the enzyme activity were assayed and the enzyme kinetic was also measured.
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