Alkaline Pectate Lyase Pretreatment for Lignocellulose Digestion

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Solution Overview

Problem

Current methods for converting cellulose into biofuels, such as ethanol, face challenges due to the recalcitrant structure of lignocellulose, requiring costly and inefficient pretreatment processes to access cellulose for enzymatic digestion, resulting in limited yield of reducing sugars.

Innovation Solution

The use of an alkaline pectate lyase with a specific amino acid sequence, or its homologues, for pretreatment of lignocellulosic materials at a pH between 9 and 12, significantly improves the yield of reducing sugars during enzymatic digestion by enhancing the accessibility of cellulose to enzymes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If chemical pretreatment is used to remove lignin barriers, then cellulose accessibility to enzymes is improved, but process cost and chemical consumption increase

Engineering Contradiction:
Improvecellulose accessibilityVSAvoidchemical consumption
Core Design Contradiction:
Ease of manufactureVSLoss of substance

Solution Approach 1:

A lignin-binding protein acts as an intermediary that selectively binds to lignin in the liquid phase, facilitating lignin removal without requiring aggressive chemical treatments. This mediator enables gentle pretreatment while achieving effective cellulose exposure for enzymatic digestion

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent replaces harsh chemical pretreatment systems with a biological/protein-based system. Instead of using acids, bases, or oxidizing agents to remove lignin barriers, the invention uses lignin-binding proteins that selectively capture lignin through specific binding interactions, substituting chemical mechanisms with biological recognition mechanisms

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Productivity

If physical size reduction is used to increase cellulose surface area, then enzymatic digestion efficiency is improved, but energy consumption increases

Engineering Contradiction:
Improveenzymatic digestion efficiencyVSAvoidenergy for size reduction
Core Design Contradiction:
ProductivityVSUse of energy by moving object

Solution Approach 1:

The lignin-binding protein performs preliminary action by selectively binding and removing lignin barriers before enzymatic digestion begins. This preliminary removal of inhibitors allows subsequent enzymatic treatment to proceed more efficiently without requiring extensive mechanical size reduction

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The lignin-binding protein serves as a mediator that facilitates the interaction between cellulose and enzymes by removing lignin interference. This mediator enables effective enzymatic digestion with minimal mechanical pretreatment, reducing the energy required for size reduction

Inventive Principle:
Principle #24Intermediary (Mediator)

3Ease of manufacture

If extensive mechanical pretreatment is applied to break lignocellulose structure, then cellulose accessibility is improved, but process complexity and equipment requirements increase

Engineering Contradiction:
Improvecellulose accessibilityVSAvoidpretreatment equipment
Core Design Contradiction:
Ease of manufactureVSDevice complexity

Solution Approach 1:

The lignin-binding protein acts as a simple yet effective intermediary that binds to lignin and facilitates its removal through filtration or separation. This approach replaces complex mechanical pretreatment equipment with a simpler protein-based solution that achieves comparable or superior cellulose accessibility

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention extracts the problematic lignin component from the lignocellulose matrix using specific binding proteins. By taking out the lignin barrier through selective binding and separation, the process achieves effective cellulose exposure without requiring complex mechanical breakdown equipment

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

This approach increases the yield of reducing sugars to up to 80-85% of the theoretical maximum, compared to previous methods, with improvements of 20-36% over standard processes, making the conversion more efficient and cost-effective.

Implementation Method 1

a pretreatment with an alkaline pectate lyase with an amino acid sequence according to SEQ ID NO: 1, greatly improves the yield of reducing sugars in a process wherein cellulose from a lignocellulosic material is enzymatically digested into reducing sugars

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

the lignocellulosic material is contacted in a pretreatment step (i.e. pretreated) with a pectate lyase at a pH between 9 and 12

Methodology Applied
Scientific EffectAlkaline hydrolysis: Hydrolysis

Data Source

PatentUS10815471B2Method for producing reducing sugar from lignocellulosic substrates
Publication Date: 2020.10.27 METGEN

AI summary

The invention relates to processes for the conversion of biomass into carbohydrates, notably reducing sugars. It provides means and methods for increasing the yield of enzymatic digestion of a biomass, in particular in methods wherein cellulose is converted into sugars using a cellulose-converting enzyme. More in particular, the invention provides a method for producing a reducing sugar from a lignocellulosic material, wherein the lignocellulosic material is contacted with a pectate lyase at a pH between 9 and 12, wherein the pectate lyase comprises an amino acid sequence according to SEQ ID NO: 1 or an amino acid sequence at least 70% identical with SEQ ID NO: 1.