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Abstract
?-Mannanase catalyzes endo-wise hydrolysis of the backbone of mannan and heteromannan, which are abundant in the cell wall structure of ungerminated leguminous seeds. The mature ?-mannanase originated from Bacillus subtilis was expressed in Pichia pastoris, a methylotrophic yeast, using the leader peptide sequence of Saccharomyces cerevisiae ?-factor. The cultivation of ?-mannanase expressing Pichia pastoris yields up to 1.8 g/L protein. In the supernatant the activity of the 40 kDa-total mannanase attained a level of 1102.0 IU/mL. The properties of the ?-mannanase were characterized. Optimum pH and temperature for the recombinant enzyme were 5.5 and 50°C respectively. The enzyme was stable at pH 5.0–10.0 and maintained over 30% original activity after incubating at 70°C for 30 min.
Keywords
β-mannanase, Pichia pastoris, expression
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Heterologous expression and characterization
of gene from in.
Front. Biol., 2008, 3(1): 26-31 DOI:10.1007/s11515-008-0002-5
