Biochemical characterization and functional analysis of invertase Bmsuc1 from silkworm, Bombyx mori.

Invertase or β-fructofuranosidase (EC 3.2.1.26) belongs to the glycoside hydrolase family 32, which catalyzes the hydrolysis of sucrose into fructose and glucose. Here, we report the biochemical and functional characterization of invertase Bmsuc1 from Bombyx mori. Bmsuc1 showed optimal hydrolysis at pH 7.0-8.0 and its optimum temperature is 50°C using sucrose as substrate. Circular dichroism spectra indicated Bmsuc1 had a primarily β-strand structure. The thermal denaturations transition of Bmsuc1 was a cooperative process with a Tm, ΔH, and ΔS of 53.81±0.12°C, 185.51±0.14KJ/mol and 0.56±0.01KJ/(molK), respectively. Moreover, homology modeling and multi-sequence alignment suggested that Bmsuc1 has a canonical β-propeller fold and one conserved catalytic triad, Asp-Asp-Glu, which is located in the bottom of the substrate-binding pocket. Bmsuc1 was expressed at high levels in the silk gland at both the transcriptional and translational levels. These expression profiles combined with invertase activity analyses of Bmsuc1 suggested that it might function as a digestive enzyme to hydrolyze sugar in the silk gland lumen. Collectively, these findings expand towards a better understanding of the structure of Bmsuc1 and its function in the silk gland.