The disclosure herein relates to a method for reducing the inhibitory effect of a cyclodextrin on a pullulanase and belongs to the technical field of gene engineering, enzyme engineering or food science. The method of the disclosure prepares a pullulanase mutant by reasonably mutating the key amino acid of the pullulanase interactive with the cyclodextrin to reduce the inhibitory effect of the cyclodextrin on the pullulanase, thereby improving the hydrolysis activity of the pullulanase. The disclosure finds the interactive sites of the pullulanase and the cyclodextrin based on analysis of crystal structures of enzymes and inhibitors and sequence comparison of enzymes from different sources, and utilizes site-directed mutation to obtain the pullulanase mutant having reduced sensibility to the cyclodextrin, thereby improving the utilization ratio of the starch raw material and the yield of the cyclodextrin.

Truncated pullulanases having an N-terminal deletion in a parental pullulanase are provided. These truncated pullulanases have altered properties as compared to the parental pullulanase, including an increased saccharification rate and higher catalytic activity at acidic pH values below 4.5 and higher temperatures of up to 64 C. These truncated pullulanase also have improved thermal stability as compared to the parental enzyme. Also provided are compositions containing the truncated pullulanases and a glucoamylase, and processes for applying the truncated pullulanases and compositions in the starch industry. Polynucleotides that encode the truncated pullulanases, and recombinant host cells for producing the truncated pullulanases are also described.

The present invention relates to a method of making glucose syrup from liquefied starch comprising, (a) contacting the liquefied starch with a glucoamylase, a pullulanase, and optionally an alpha-amylase wherein the ratio of pullulanase dose expressed as NPUN/gDS, to alpha-amylase dose expressed as FAU(A)/gDS is at least 60, particularly at least 75, particularly at least 100, more particularly at least 150, more particularly at least 200, more particularly at least 250, more particularly at least 300, more particularly at least 400, more particularly at least 500, more particularly at least 600, more particularly at least 800 or if no alpha-amylase is present the pullulanse is present in a dose of at least 0.5, particularly at least 0.75, particularly at least 1.0, particularly at least 1.5 NPUN/gDS, and (b) saccharifying the liquefied starch.

The present disclosure relates to tagatose-6-phosphate phosphatase consisting of an amino acid sequence of SEQ ID NO: 1, a nucleic acid encoding the tagatose-6-phosphate phosphatase, and a transformant comprising the nucleic acid. Additionally, the present disclosure relates to a composition for producing tagatose, which comprises the tagatose-6-phosphate phosphatase of the present disclosure, and a method for producing tagatose using the tagatose-6-phosphate phosphatase of the present disclosure.

The present invention relates to a method of making glucose syrup from liquefied starch comprising, (a) contacting the liquefied starch with a glucoamylase, a pullulanase, and optionally an alpha-amylase wherein the ratio of pullulanase dose expressed as NPUN/gDS, to alpha-amylase dose expressed as FAU(A)/gDS is at least 60, particularly at least 75, particularly at least 100, more particularly at least 150, more particularly at least 200, more particularly at least 250, more particularly at least 300, more particularly at least 400, more particularly at least 500, more particularly at least 600, more particularly at least 800 or if no alpha-amylase is present the pullulanse is present in a dose of at least 0.5, particularly at least 0.75, particularly at least 1.0, particularly at least 1.5 NPUN/gDS, and (b) saccharifying the liquefied starch.

The present invention relates to thermostable pullulanases useful for industrial and scientific purposes. The present invention provides methods for producing the modified pullulanase, enzymatic compositions comprising the modified pullulanase, and methods for use of the enzymatic compositions.

The present disclosure relates to liquid enzyme formulations containing one or more alpha-amylases for use in starch processing, wherein the pH of the enzyme formulation is about pH 6.0-8.0, and methods of use thereof. The present disclosure further relates to methods of making a liquid enzyme formulation containing one or more alpha-amylase having improved stability, comprising titrating the pH of the liquid enzyme formulation to a range of pH 6.0-8.0.

A method of mashing comprising providing a grist comprising malt and adjunct; and contacting the grist with a pullulanase; an alpha amylase; and a maltogenic alpha amylase and/or a beta amylase to make a wort. An enzyme composition comprising a pullulanase; an alpha amylase; and a maltogenic alpha amylase and/or a beta amylase and the use of these enzymes in brewing is disclosed.

The present invention relates to thermostable pullulanases useful for industrial and scientific purposes. The present invention provides methods for producing the modified pullulanase, enzymatic compositions comprising the modified pullulanase, and methods for use of the enzymatic compositions.

The present disclosure relates to liquid enzyme formulations containing one or more alpha-amylases for use in starch processing, wherein the pH of the enzyme formulation is about pH 6.0-8.0, and methods of use thereof. The present disclosure further relates to methods of making a liquid enzyme formulation containing one or more alpha-amylase having improved stability, comprising titrating the pH of the liquid enzyme formulation to a range of pH 6.0-8.0.