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Characterization of a novel low-temperature-active, alkaline and sucrose-tolerant invertase
- Source :
- Scientific Reports
- Publication Year :
- 2016
- Publisher :
- Springer Science and Business Media LLC, 2016.
-
Abstract
- A glycoside hydrolase family 32 invertase from Bacillus sp. HJ14 was expressed in Escherichia coli. The purified recombinant enzyme (rInvHJ14) showed typical biochemical properties of low-temperature-active and alkaline enzymes: (i) rInvHJ14 was active and stable in the range of pH 7.0–9.5 with an apparent pH optimum of 8.0; (ii) rInvHJ14 was most active but not stable at 30–32.5 °C, with 19.7, 48.2 and 82.1% of its maximum activity when assayed at 0, 10 and 20 °C, respectively and the Ea, ΔG* (30 °C), Km (30 °C) and kcat (30 °C) values for hydrolysis of sucrose by rInvHJ14 was 47.6 kJ mol−1, 57.6 kJ mol−1, 62.9 mM and 746.2 s−1, respectively. The enzyme also showed strong sucrose tolerance. rInvHJ14 preserved approximately 50% of its highest activity in the presence of 2045.0 mM sucrose. Furthermore, potential factors for low-temperature-active and alkaline adaptations of rInvHJ14 were presumed. Compared with more thermostable homologs, rInvHJ14 has a higher frequency of glycine residues and a longer loop but a lower frequency of proline residues (especially in a loop) in the catalytic domain. The catalytic pockets of acid invertases were almost negatively charged while that of alkaline rInvHJ14 was mostly positively charged.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Sucrose
Glycosylation
Protein Conformation
Glycine
Bacillus
01 natural sciences
Article
Substrate Specificity
03 medical and health sciences
chemistry.chemical_compound
Hydrolysis
Sequence Analysis, Protein
Catalytic Domain
Escherichia coli
Glycoside hydrolase
Enzyme kinetics
Proline
chemistry.chemical_classification
Multidisciplinary
beta-Fructofuranosidase
Temperature
Hydrogen-Ion Concentration
Recombinant Proteins
Kinetics
030104 developmental biology
Enzyme
Invertase
chemistry
Biochemistry
Thermodynamics
010606 plant biology & botany
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....29b723fbfae01a22e2ac526eb9e1b102