Enzymatic activity and substrate specificity of the recombinant tomato β-galactosidase 1

• Published in: Journal of plant physiology Vol. 171; no. 16; pp. 1454 - 1460
• Main Authors: Eda, Masahiro, Ishimaru, Megumi, Tada, Toshiji, Sakamoto, Tatsuji, Kotake, Toshihisa, Tsumuraya, Yoichi, Mort, Andrew J., Gross, Kenneth C.
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 15.10.2014
• Subjects: beta-Galactosidase - genetics; beta-Galactosidase - metabolism; Cell wall; Correlation; Exo-galactanase; Frames; Fruit - metabolism; Fruits; Galactose; Galactose - metabolism; Lupins; Lycopersicon esculentum; Lycopersicon esculentum - enzymology; Lycopersicon esculentum - genetics; Lycopersicon esculentum - metabolism; Plant Proteins - genetics; Plant Proteins - metabolism; Recombinant; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Ripening; Saccharomyces cerevisiae - genetics; Solanum lycopersicum; Substrate Specificity; Walls; β-Galactosidase; CSP; Dap; Solanum lycopersicum; Substrate specificity; Exo-galactanase; TBG; ASP; HF; Cell wall; β-Galactosidase; ABEE
• ISSN: 0176-1617; 1618-1328
• DOI: 10.1016/j.jplph.2014.06.010

The open reading frame of tomato β-galactosidase 1 was expressed in yeast, and the enzymatic properties and substrate specificity were investigated. The enzyme had peak activity at pH 5.0 and 40–50°C. TBG1 was active on β-(1,3)- and β-(1,6)-galactobiose and lactose. TBG1 released galactose from lupin galactan, tomato fruit alkali soluble pectin, arabinogalactan, gum arabic and methyl β-(1,6)-galactohexaoside, but not from labeled β-(1,4)-galactoheptaose. TBG1 was assessed for its ability to degrade three galactosyl-containing cell wall fractions purified from different development and ripening stages of tomato fruit. TBG1 released galactose from all of the fractions from all of the stages tested. TBG1 activity was highest on the hemicellulose fraction at the 10 and 20d after pollination stage. This result is not correlated the with TBG1 expression pattern. TBG1 might act on a small but specific set of polysaccharide containing galactose.