We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Purification and Characterization of Exo-Inulinase from <italic>Paenibacillus</italic> sp. d9 Strain.
- Authors
Jeza, S.; Maseko, S. B.; Lin, J.
- Abstract
This study intended to purify and characterise exo-inulinase of diesel-degrading <italic>Paenibacillus</italic> sp. D9. The whole genome sequencing of <italic>Paenibacillus</italic> sp. D9 revealed to possess the <italic>sacC</italic> gene that is encoded as exo-inulinase/levanase. This isolate was capable of producing a maximum of 50.9 IU/mL of exo-inulinase activity within 3 days at 30 °C, 200 rpm and pH of 7.0 on minimal salt medium agar supplemented with 1% (w/v) inulin. An exo-inulinase of 58.5 kDa was purified using ammonium sulphate precipitation, HiTrap QFF column and MMC column chromatographies with a specific activity of 4333 IU/mg, 7.1% recovery and a 4.3-fold increase in purity. The purified D9 exo-inulinase had temperature and pH optimum at 40 °C and pH 4.0, respectively, with the Michaelis constant of 5.5 mM and a maximal velocity of 476.2 IU/mg, respectively. Catalytic constant, <italic>k</italic>cat was calculated to be 42.6 s−1 with a catalytic efficiency (<italic>k</italic><italic>cat</italic>/<italic>K</italic><italic>m</italic>) of 7.6 s−1 mM−1. The presence of Ca2+ enhanced the activity of D9 exo-inulinase while Hg2+ completely inhibited the activity, other compounds such as Fe3+ and Cu2+ had an inhibitory effect. The results of amino acid alignment and the complete degradation of inulin into fructose by the purified enzyme confirmed that inulinase from <italic>Paenibacillus</italic> sp. D9 is an exo-form. The phylogenetic tree based on the protein sequences indicates that bacterial exo-inulinases possess a common ancestry.
- Subjects
PAENIBACILLUS; INULASE; AMINO acid sequence; NUCLEOTIDE sequencing; BACTERIAL protein analysis
- Publication
Protein Journal, 2018, Vol 37, Issue 1, p70
- ISSN
1572-3887
- Publication type
Article
- DOI
10.1007/s10930-017-9752-8