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AssigneeUniversiteit Gent
InventorsJef Van Der Borght, Tom Desmet, Wim Soetaert, Chao Chen
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The present invention relates to trehalose phosphorylases which are useful for the industrial production of trehalose-analogues and glycosyl phosphates. More specifically, the invention discloses trehalose phosphorylases which are mutated in specific amino acid regions. These specific mutations result in modified substrate specificities of the enzymes. In addition, the present invention discloses a wild type trehalose phosphorylase from the marine organism Caldanaerobacter subterraneus, and mutated types thereof, which are highly thermostable and have a broad acceptor and donor specificity.
A mutated trehalose phosphorylase containing at least one mutation in the amino acid positions 371, 442, 450, 649, 659 or 693 wherein said amino acid positions correspond to the amino acid positions as determined by SEQ ID N° 1 or to corresponding amino acid positions in a trehalose phosphorylase having an amino acid sequence which is at least 75% identical to SEQ ID N° 1, and wherein said mutated trehalose phosphorylase has an increased catalytic efficiency towards its acceptor or donor substrate compared to the corresponding wild-type trehalose phosphorylase.
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A variant trehalose phosphorylase characterized in having the amino acid sequence as depicted by SEQ ID N° 1 but having at least one mutation in the amino acid regions 330-410 or 630-710 so that, compared to the wild type enzyme, said variant displays a significant increase in the ratio of beta-galactose-1-phosphate over beta-glucose-1-phosphate release from lactotrehalose.
A method to produce a glycosyl phosphate comprising:
Dependencies are inferred from the claim text.