Phospholipase C Enzyme Safety and pH Stability

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Solution Overview

Problem

Conventional phospholipase C enzymes from animals, bacteria, and fungi lack the necessary properties for efficient use in the food and oil mill industries, such as broad substrate specificity, stability across pH ranges, and safety concerns, particularly due to pathogenic issues and phosphatase activity.

Innovation Solution

Purified phospholipase C enzymes derived from Aspergillus oryzae strain FERM BP-10200 or Aspergillus tamarii strain IAM 13907, which exhibit activity from acidic to neutral pH, are nonspecific to phosphatidylinositol, have optimal pH between pH 3 to pH 6, and show stability at temperatures up to 80°C and pH 4.5, while avoiding phosphate ester hydrolysis except for phospholipids.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional phospholipase C enzymes from animals, bacteria, and fungi are used, then phospholipid hydrolysis activity is achieved, but safety concerns arise due to pathogenic issues and phosphatase activity

Engineering Contradiction:

ImprovesafetyVSAvoidpathogenic issues and phosphatase activity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and isolates the specific phospholipase C gene from non-pathogenic Aspergillus species, separating the desired enzymatic function from the harmful pathogenic properties associated with conventional sources. This extraction of the functional gene allows production of safe enzymes without phosphatase activity.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent employs Aspergillus species that are non-pathogenic and can be safely cultured, replacing dangerous pathogenic sources. The use of safe, easily culturable fungi provides a reliable and safe source for enzyme production without the risks associated with pathogenic organisms.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

2Productivity

If phospholipase C enzymes with broad substrate specificity are used, then efficiency in hydrolyzing various phospholipids is improved, but selectivity is reduced

Engineering Contradiction:

Improveefficiency in hydrolyzing various phospholipidsVSAvoidsubstrate selectivity

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent modifies the enzymatic properties by selecting and characterizing phospholipase C variants from Aspergillus species that exhibit optimal activity across different pH ranges (acidic to neutral) while maintaining broad substrate specificity. The enzymes show activity against phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol with balanced efficiency and selectivity.

Inventive Principle:

Principle #35Parameter changes

3Adaptability or versatility

If phospholipase C enzymes stable across wide pH ranges are used, then applicability in food and oil mill industries is improved, but enzyme stability is compromised

Engineering Contradiction:

Improveapplicability in food and oil mill industriesVSAvoidenzyme stability

Core Design Contradiction:

Adaptability or versatilityVSStability of the object's composition

Solution Approach 1:

The patent identifies and utilizes phospholipase C enzymes from Aspergillus species that exhibit pH-dependent activity profiles optimized for specific industrial applications. The enzymes maintain stable structure and activity in the pH ranges relevant to food and oil mill industries, with optimal activity at acidic to neutral pH values while remaining structurally stable.

Inventive Principle:

Principle #3Local quality

4Reliability

If phospholipase C enzymes with heat stability are used, then process robustness is improved, but activity at lower temperatures may be reduced

Engineering Contradiction:

Improveheat stabilityVSAvoidactivity temperature range

Core Design Contradiction:

ReliabilityVSTemperature

Solution Approach 1:

The patent characterizes phospholipase C enzymes from Aspergillus species that exhibit dynamic temperature adaptation, maintaining structural stability at elevated temperatures while retaining catalytic activity across a broad temperature range. The enzymes demonstrate optimal activity at moderate temperatures with increased stability at higher temperatures, allowing flexible application in various industrial processes.

Inventive Principle:

Principle #15Dynamics

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

These enzymes demonstrate enhanced safety and efficiency in hydrolyzing various phospholipids across acidic and neutral pH ranges, maintaining activity in citrate buffer solutions, and showing heat stability, making them suitable for food and oil mill applications without degrading phosphate esters.

Implementation Method 1

phospholipase C hydrolyzes glycerophospholipid into diacylglycerol and a phosphoryl base

Methodology Applied

Scientific EffectEnzymatic hydrolysis: Enzyme

Implementation Method 2

Phospholipase C hydrolyzes a phospholipid into diacylglycerol and a phosphoryl base

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS7993874B2Phospholipase C enzyme(s)

Publication Date: 2011.08.09 MITSUBISHI CHEM CORP

AI summary

The present invention provides a phospholipase C enzyme(s) having ability to hydrolyze phospholipid in both acidic and around neutral ranges and the activity in a citrate buffer solution as well as having some degree of heat stability, and having a property not to hydrolyze phosphate esters not containing lipid moieties. The phospholipase C enzyme(s) shows the activity at from acidic to neutral pH and does not substantially hydrolyze any phosphate esters except for phospholipids.