Prokaryotic expression and characterization of a keratinolytic protease from Aspergillus niger
-
Xiaoling Chen
,
Hua Zhao
Abstract
In this study, a keratinolytic protease gene (named as kerD) from Aspergillus niger was cloned. The full-length coding sequence of kerD consists of 1,251 bp and encodes a protein with 416 amino acid residues with a molecular mass of 43,831 Da. A DNA fragment encoding mature kerD without its signal sequence was inserted into the expression vector pET30a(+) and successfully expressed in Escherichia coli. The recombinant protein was purified to approximately 100% purity using Ni-IDA affinity chromatography, and identified by Western blot. The recombinant enzyme had an optimal pH of 8.0 and was stable at pH 7.0-9.0. It exhibited an optimal temperature for activity of 70 °C and was stable at 30-50 °C. It was highly inhibited by 1,10-phenanthroline, ethylenediaminetetraacetic acid and sodium dodecyl sulphate, but activated by phenylmethanesulfonyl fluoride, Mg2+, Fe2+, Mn2+, Cu2+, Zn2+, Ca2+, dithiothreitol, Triton X-100, dimethyl sulfoxide and isopropyl alcohol. The recombinant enzyme could hydrolyse a broad range of protein substrates.
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Artikel in diesem Heft
- Biodiversity and screening of halophilic bacteria with hydrolytic and antimicrobial activities from Yuncheng Salt Lake, China
- Prokaryotic expression and characterization of a keratinolytic protease from Aspergillus niger
- ABA-dependent sucrose regulation of antioxidant metabolism in wheat cultivars varying in ABA-sensitivity
- Identification of salt tolerant Acacia species for saline land utilisation
- Phylogenetic relationships among Elymus and related diploid genera (Triticeae: Poaceae) based on nuclear rDNA ITS sequences
- Adaptations to oxidative stress in Zea mays roots under short-term Pb2+ exposure
- Zinc alleviates cadmium effects on growth, membrane lipid biosynthesis and peroxidation in Solanum lycopersicum leaves
- Comparison of the floodplain forest floristic composition of two riparian corridors: species richness, alien species and the effect of water regime changes
- Red list of ferns and flowering plants of Slovakia, 5th edition (October 2014)
- Distribution and abundance of Hemolivia mauritanica (Apicomplexa: Haemogregarinidae) and its vector Hyalomma aegyptium in tortoises of Iran
- First record of mite genus Ledermuelleriopsis (Acari: Stigmaeidae) in South America with redescription of L. verricula from Brazil
- The influence of agricultural management on the structure of ground beetle (Coleoptera: Carabidae) assemblages
- Insect community on Jurinea cyanoides (Asteraceae), a plant species protected under NATURA 2000
- Chlorella pyrenoidosa, young barley and fruit peel polyphenols in rat breast cancer model – the effects on plasma lipid metabolism
- Increased endogenous serotonin level in diabetic conditions may lead to cardiac valvulopathy via reactive oxygen species regulation
- Parasitic helminths in grey heron (Ardea cinerea) chicks
