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Polypeptide with glucoamylase activity and preparation method thereof
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A technology of glucoamylase and activity, applied in the field of polypeptides
Active Publication Date: 2017-12-26
DUPONT NUTRITION BIOSCIENCES APS
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The reason this method is not commonly used is that active enzyme can then be present in the final product, which is undesirable as stated above
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Embodiment 1
[0263] Example 1. An isolated polypeptide having glucoamylase activity, the polypeptide being selected from:
[0264] a) A polypeptide comprising an amino acid sequence that has at least 75%, 76%, 77%, 78%, 79%, 80%, 81% of the amino acid sequence of a mature polypeptide selected from SEQ ID NOs: 3 and 6 , 82%, 83%, 84%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity;
[0265] b) A polypeptide encoded by: i) the nucleic acid sequence contained in SEQ ID NO: 1 or SEQ ID NO: 4, or ii) the cDNA sequence of i), or iii) SEQ ID NO: 2 or SEQ ID NO: The sequence of 5, or iv) a polynucleotide that hybridizes to the complementary strand of i), ii), or iii) under at least low stringency conditions;
[0266] c) A polypeptide comprising conservative substitutions, deletions and / or insertions of one or more amino acids in an amino acid sequence selected from the mature polypeptides of SEQ ID NO: 3 and 6;
[0269] Example 2. An isolated polypeptide with glucoamylase activity, said polypeptide comprising an amino acid sequence that is at least 75%, 76%, 77% of the amino acid sequence of a mature polypeptide selected from SEQ ID NO: 3 and 6 , 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity.
Embodiment 3
[0270] Example 3. An isolated polypeptide having glucoamylase activity, said polypeptide being encoded by: i) the nucleic acid sequence contained in SEQ ID NO: 1 or SEQ ID NO: 4, or ii) i) cDNA sequence, or iii) the sequence of SEQ ID NO: 2 or SEQ ID NO: 5, or iv) a polynucleotide that hybridizes to the complementary strand of i), ii), or iii) under at least low stringency conditions.
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Abstract
The present invention relates to polypeptides having glucoamylase activity with improved properties and to compositions comprising these polypeptides suitable for use in the production of a food, beverage (e.g. beer), feed, or biofuel. Also described is an improved and cost-effective process for isolating glucoamylases suitable for large scale protein purification procedures. Furthermore, different methods and uses related to glucoamylases according to the invention are disclosed, such as in a brewing process.
Description
[0001] Reference to Sequence Listing [0002] Attached is a sequence listing including SEQ ID NO: 1-17, which is incorporated herein by reference in its entirety. Technical field [0003] The present invention relates to polypeptides with glucoamylase activity and improved properties, and to compositions containing these polypeptides suitable for the production of food, beverages (such as beer), feed or biofuels. An improved and cost-effective method for isolating glucoamylase suitable for large-scale protein purification procedures is also described. In addition, different methods and uses related to the glucoamylase according to the present invention are disclosed, for example in the brewing process. Background technique [0004] Glucoamylase (glucan 1,4-α-glucosylhydrolase, EC 3.2.1.3) is a starch hydrolyzing exo-acting carbohydrase, which catalyzes the removal from the non-reducing end of starch or related oligosaccharide and polysaccharide molecules. Divide consecutive glucos...
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