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High‐level expression of a neoagarobiose‐producing β‐agarase gene from Agarivorans sp. JAMB‐A11 in Bacillus subtilis and enzymic properties of the recombinant enzyme

The structural gene for a neoagarobiose‐producing β‐agarase of an Agarivorans isolate was expressed in Bacillus subtilis. High‐level production of the recombinant enzyme was achieved corresponding to a level of 1.9×104 units/l of the culture broth. The efficiency of the production is thus 30‐fold greater than that of the original strain. The recombinant enzyme (RagaA11) had a molecular mass of 105 kDa and a specific activity of 371 units/mg. The optimal pH and temperature of the enzyme were 7.5–8.0 and 40 °C respectively. The enzyme is an endo‐type β‐agarase hydrolysing not only agarose, but also neoagarotetraose, to yield neoagarobiose as the final main product, representing approx. 90 mol% of total products. RagaA11 would be useful for industrial production of neoagarobiose.