Modified Xylanase Polypeptides for Bran Solubilization

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current xylanases used in various applications often have low catalytic activity, making them inefficient for commercial use, particularly in cereal bran processing and cellulosic bio-ethanol production, where high xylanase activity and bran solubilization are required.

Innovation Solution

Modifying the B. subtilis xylanase polypeptide sequence by introducing specific amino acid substitutions at positions 12, 13, and others (15, 34, 54, 77, 81, 82, 99, 104, 110, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166, 175, 179) to enhance xylanase activity and bran solubilization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If current xylanases are used in commercial applications, then the process is simple and straightforward, but the catalytic activity is low making them inefficient

Engineering Contradiction:
Improvexylanase activityVSAvoidpolypeptide structure complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent applies parameter changes by modifying specific amino acid residues in the xylanase polypeptide sequence. Multiple amino acid substitutions are introduced at positions including 12, 13, 15, 34, 54, 77, 81, 82, 99, 104, 110, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166, 175, and 179. These parameter changes at the molecular level result in significantly enhanced catalytic activity and bran solubilization while maintaining the fundamental enzyme structure and function.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If more xylanase is used to achieve sufficient activity, then the catalytic performance improves, but the cost and complexity of the process increases

Engineering Contradiction:
Improvexylanase activityVSAvoidamount of xylanase needed
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The patent achieves higher productivity with reduced quantity by modifying the catalytic efficiency of individual xylanase molecules through amino acid substitutions. The modified polypeptides exhibit enhanced catalytic activity per unit mass, meaning less enzyme is required to achieve the same level of xylan hydrolysis and bran solubilization, thereby reducing material costs while improving process efficiency.

Inventive Principle:
Principle #35Parameter changes

3Productivity

If xylanase activity is increased through modification, then the efficiency in cereal bran processing improves, but the manufacturing complexity of the modified polypeptide increases

Engineering Contradiction:
Improvebran solubilization efficiencyVSAvoidpolypeptide production ease
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent balances manufacturing ease with improved performance by implementing targeted amino acid substitutions rather than comprehensive restructuring. The modifications are introduced through site-directed mutagenesis or recombinant DNA technology, allowing for precise control over the number and position of substitutions. This approach enables improved bran solubilization efficiency while maintaining relatively simple manufacturing processes through established biotechnological methods.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies segmentation by dividing the polypeptide structure into specific regions and positions for modification. Rather than attempting to modify the entire enzyme globally, the invention focuses on specific amino acid positions (12, 13, 15, 34, 54, 77, 81, 82, 99, 104, 110, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166, 175, 179) that are most critical for catalytic activity and substrate interaction. This segmented approach to modification simplifies the manufacturing process by focusing resources on key modification sites.

Inventive Principle:
Principle #1Segmentation

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 modified xylanases exhibit increased xylanase activity and bran solubilization, enabling more efficient hydrolysis of hemicellulosic fractions in cereal processing and reducing the amount of xylanase needed in applications like bio-ethanol production, animal feed, and bakery industries.

Implementation Method 1

endo-β-1,4-xylanases (EC 3.2.1.8) (referred to herein as xylanases) have been used for the modification of complex carbohydrates derived from plant cell wall material

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

The modified xylanases exhibit increased xylanase activity and bran solubilization, enabling more efficient hydrolysis of hemicellulosic fractions in cereal processing

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP2382310B1Polypeptides with xylanase activity
Publication Date: 2016.08.24 INT N&H DENMARK APS
  • EP2382310B1 patent drawingFigure 1
  • EP2382310B1 patent drawingFigure 2
  • EP2382310B1 patent drawingFigure 2

AI summary

Polypeptides with xylanase activity modified to increase bran solubilization and/or xylanase activity. The modification comprises modification of one or more amino acids in position 12 or 13 in combination with one or more further amino acid modifications in position 15, 34, 54, 77, 81, 82, 99, 104, 110, 113, 114, 118, 122, 141, 154, 159, 162, 164, 166, 175 or 179, wherein the positions are determined as the position corresponding the position of Bacillus subtilis xylanase (SEQ ID NO 1).