Use of alpha-1,4-glucan lyase for preparation of 1,5-D-anhydrofructose
AssigneeDanisco A/S
InventorsShukun Yu, Kirsten Bojsen, Karsten Kragh, Maja Bojko, John Nielsen, Jan Marcussen, Tove Christensen
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Abstract
A method of preparing the sugar 1,5-D-anhydrofructose is described. The method comprises treating an α-1,4-glucan with an α-1,4-glucan lyase wherein the enzyme is used in substantially pure form. In a preferred embodiment, if the glucan contains links other than and in addition to the α-1,4-links, the α-1,4-glucan lyase is used in conjunction with a suitable reagent that can break the other links.
Claims22
A method of preparing the sugar 1,5-D-anhydrofructose comprising treating an α-1,4-glucan with the enzyme α-1,4-glucan lyase characterised in that enzyme is used in substantially pure form.
A method according to any preceding claim wherein the α-1,4-glucan lyase is bound to a support or, more preferably, is in a dissolved form.
A method according to any preceding claim wherein the enzyme is isolated from either a fungus, preferably Morchella costata or Morchella vulgaris, or from a fungally infected algae, preferably Gracilariopsis lemaneiformis or from algae alone, preferably Gracilariopsis lemaneiformis.
A method according to any of the preceding claims wherein the enzyme comprises the amino acid sequence SEQ. ID. No. 1. or the amino acid sequence SEQ. ID. No. 2 or the amino acid sequence SEQ. ID. No. 5. or the amino acid sequence SEQ. ID. No. 6, or any variant thereof.
A method according to any preceding claim wherein the enzyme is obtained from the expression of a nucleotide sequence coding for the enzyme.
The method according to any one of the preceding claims wherein the substrate is treated with the enzyme in the presence of a buffer.
The method according to any one of the preceding claims wherein the substrate is treated with the enzyme in the absence of a co-factor.
The method according to any one of the preceding claims wherein the enzyme is used in combination with amylopectin or dextrin.
A method of preparing the sugar 1,5-D-anhydrofructose comprising treating an α-1,4-glucan with the enzyme α-1,4-glucan lyase characterised in that enzyme comprises the amino acid sequence SEQ. ID. No. 1. or the amino acid sequence SEQ. ID. No. 2 or the amino acid sequence SEQ. ID. No. 5. or the amino acid sequence SEQ. ED. No. 6, or any variant thereof.
The sugar 1,5-D-anhydrofructose when prepared by the method of the present invention.
The use of a reagent that can increase the hydrophobicity of the reaction medium to increase the stability and activity of the GL enzyme.
Use of AF as an anti-oxidant.
Use of AF as a sweetener.
Dependencies are inferred from the claim text.