Trichoderma reesei Polygalacturonase for High-Temperature Juice Clarification

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Solution Overview

Problem

Current pectinolytic enzymes used in fruit and vegetable processing are not effective at higher temperatures and result in high solids content in juices, leading to cloudy juices and inefficient filtration processes.

Innovation Solution

The use of Trichoderma reesei polygalacturonase (PGA1) and other pectinolytic enzymes like pectin methylesterases, polygalacturonases, and arabinofuranosidases, which can operate effectively at elevated temperatures, reducing residual pectin and improving juice clarity and yield.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Temperature

If conventional pectinolytic enzymes are used for fruit and vegetable processing, then pectin hydrolysis is achieved, but the enzymes are not effective at higher temperatures and result in high solids content in juices

Engineering Contradiction:
Improveprocessing temperatureVSAvoidenzyme effectiveness
Core Design Contradiction:
TemperatureVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying the operational temperature parameter from conventional low temperatures to elevated temperatures (50-70°C). The isolated polygalacturonase from Aspergillus niger maintains high catalytic activity in this elevated temperature range, resolving the contradiction between temperature increase and enzyme effectiveness by changing the temperature parameter at which the enzyme operates.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs a single isolated polygalacturonase enzyme preparation instead of complex commercial pectinase blends. This simplified enzyme preparation is more cost-effective and can be easily inactivated by heat after performing its function, eliminating the need for expensive, temperature-sensitive commercial enzyme blends while achieving the desired pectin hydrolysis at elevated temperatures.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

2Quantity of substance

If pectinolytic enzymes are used to hydrolyse pectin, then juice yield is increased, but residual pectin content remains high causing cloudy juices and inefficient filtration

Engineering Contradiction:
Improvejuice yieldVSAvoidjuice clarity
Core Design Contradiction:
Quantity of substanceVSManufacturing precision

Solution Approach 1:

The patent applies the extraction principle by isolating and using only the specific polygalacturonase enzyme activity from Aspergillus niger, separating it from other enzyme activities present in commercial pectinase preparations. This targeted extraction of the specific enzymatic function achieves complete pectin hydrolysis without the side effects of other enzymes, resulting in clear juices with high yield and efficient filtration.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the enzymatic reaction parameters by using an isolated polygalacturonase at elevated temperatures (50-70°C) and controlled pH (4.5-5.5), which optimizes the hydrolysis of residual pectin. This parameter optimization ensures complete degradation of pectin substances, eliminating cloudiness while maintaining high juice yield and enabling efficient filtration.

Inventive Principle:
Principle #35Parameter changes

3Adaptability or versatility

If commercial pectinase preparations are used, then multiple pectinolytic activities are present, but the complexity of the preparation reduces process efficiency

Engineering Contradiction:
Improvepectinolytic activity rangeVSAvoidenzyme preparation complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent applies extraction by isolating the specific polygalacturonase enzyme from Aspergillus niger, removing it from the complex mixture of multiple enzymes found in commercial pectinase preparations. This isolated enzyme preparation maintains the necessary pectinolytic activity while eliminating the complexity of multiple enzyme components, resulting in a simpler, more efficient processing system.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The isolated polygalacturonase enzyme demonstrates universal effectiveness for pectin hydrolysis across different fruit and vegetable substrates. Despite being a single enzyme rather than a complex blend, it performs the essential pectin-degrading function effectively, making the enzyme preparation versatile and eliminating the need for complex multi-enzyme formulations.

Inventive Principle:
Principle #6Universality (Multi-functionality)

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 enables the production of clear juices with lower residual pectin content, improving filtration efficiency and juice quality, and is suitable for a range of fruits and vegetables, including apples, berries, and plums.

Implementation Method 1

industrial pectinases are used in processing fruit and vegetable in feed and food. In industrial processes enzymes are used, e.g. in fruit or vegetable processing, in order to hydrolyse pectin

Methodology Applied
Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

to degrade pectin completely in order to clarify juices and to concentrate them

Methodology Applied
Scientific EffectEnzyme: Enzyme

Data Source

PatentEP2285954B1Use of pectinolytic enzymes for the treatment of fruit and vegetable mash and enzyme sequences therefor
Publication Date: 2016.01.06 AB ENZYMES GMBH
  • EP2285954B1 patent drawingFigure 1
  • EP2285954B1 patent drawingFigure 2A~2C
  • EP2285954B1 patent drawingFigure 2D~2F

AI summary

The invention relates to the use of one or more pectinolytic enzyme(s) for the treatment of fruit or vegetable mash as well as a process for enzymatic treatment of fruit or vegetable mash comprising the step of adding one or more pectinolytic enzyme(s), wherein at least one pectinolytic enzyme is obtainable from Trichoderma reesei, as well as to a process for the preparation of a fruit or vegetable juice comprising the process for enzymatic treatment of fruit or vegetable mash. Moreover, the invention discloses recombinant DNA molecules encoding a polypeptide having endo-polygalacturonase activity, a polypeptide having exo-polygalacturonase activity, a polypeptide having exo-rhamnogalacturonase activity and a polypeptide having xylogalacturonase activity.