Alpha-amylase Polypeptides for Starch Conversion Efficiency

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Solution Overview

Problem

Current alpha-amylase enzymes used in starch processing and industrial applications face limitations in efficiency and specificity, particularly in starch liquefaction, textile washing, and ethanol production, where they may not fully convert starch into maltodextrins and mono- and disaccharides effectively.

Innovation Solution

Development of isolated polypeptides with specific sequence identities and carbohydrate binding domains that enhance alpha-amylase activity, allowing for efficient conversion of starch into maltodextrins and mono- and disaccharides, suitable for starch liquefaction, textile washing, and ethanol production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If current alpha-amylase enzymes are used in starch processing, then the basic starch conversion function is achieved, but the efficiency and specificity are insufficient

Engineering Contradiction:

Improvestarch conversion efficiencyVSAvoidconversion specificity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent segments the alpha-amylase enzyme into two independent domains: a catalytic domain (residues 1-497) and a carbohydrate-binding domain (residues 498-627). This segmentation allows each domain to be optimized independently, with the catalytic domain responsible for hydrolysis efficiency and the carbohydrate-binding domain responsible for substrate specificity and binding affinity, thereby resolving the contradiction between productivity and reliability.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent applies local quality by assigning different functional properties to different regions of the enzyme. The catalytic domain is optimized for hydrolytic activity with specific amino acid residues (Asp197, Glu233, Asp300) positioned for optimal catalysis, while the carbohydrate-binding domain is optimized for specific starch substrate recognition. This localized functional optimization enables high conversion efficiency while maintaining high specificity.

Inventive Principle:

Principle #3Local quality

2Productivity

If conventional alpha-amylase is used in ethanol production, then starch hydrolysis occurs, but the conversion to maltodextrins and sugars is not complete

Engineering Contradiction:

Improvestarch to sugar conversion rateVSAvoidproduct composition control

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent utilizes parameter changes by optimizing specific amino acid residues in the catalytic domain (Asp197, Glu233, Asp300) to alter the enzyme's catalytic parameters. These parameter changes enable the enzyme to achieve complete starch hydrolysis while controlling the product distribution to maximize maltodextrin and sugar yield, thereby improving both conversion rate and product composition control.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite functional structure by combining the catalytic domain with the carbohydrate-binding domain in a single polypeptide chain. This composite structure integrates both catalytic activity and substrate binding specificity, enabling complete and controlled starch conversion to desired products for ethanol production.

Inventive Principle:

Principle #40Composite materials

3Productivity

If traditional alpha-amylase is used in textile washing, then starch removal is achieved, but the process efficiency is limited

Engineering Contradiction:

Improvestarch removal speedVSAvoidenzyme production cost

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent extracts and optimizes the essential catalytic function by focusing on the catalytic domain (residues 1-497) with its key residues (Asp197, Glu233, Asp300). This extraction of the core functional element allows for simplified production and expression of the enzyme while maintaining high starch removal efficiency, thereby improving productivity and reducing manufacturing complexity.

Inventive Principle:

Principle #2Taking out (Extraction)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The polypeptides demonstrate improved alpha-amylase activity, enabling effective starch conversion and utilization in various industrial processes, including ethanol production and textile processing, with enhanced efficiency and specificity.

Implementation Method 1

Alpha-amylases (alpha-1,4-glucan-4-glucanohydrolases, EC. 3.2.1.1) constitute a group of enzymes which catalyze hydrolysis of starch and other linear and branched 1,4-glucosidic oligo- and polysaccharides.

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

The present invention relates to isolated polypeptides having alpha-amylase activity... catalyze hydrolysis of starch

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentUS11788078B2Processes for producing ethanol

Publication Date: 2023.10.17 NOVOZYMES AS

AI summary

The present invention relates to isolated polypeptides having alpha-amylase activity, catalytic domains, carbohydrate binding domains and polynucleotides encoding the polypeptides, catalytic domains or carbohydrate binding domains. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides, catalytic domains or carbohydrate binding domains.