Genetically Engineered Candida utilis for Kitchen Waste Ethanol Production

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Solution Overview

Problem

Candida utilis lacks enzymes to effectively degrade starch and proteins in kitchen waste, limiting its ability to use these as carbon and nitrogen sources for ethanol production through fermentation.

Innovation Solution

Integration of alpha-amylase, glucoamylase, and acid protease genes from Aspergillus oryzae and Aspergillus niger into the Candida utilis genome using a multigene co-expression vector, with surface display and secretory expressions to enable degradation of starch and proteins in kitchen waste.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If Candida utilis is used for ethanol fermentation, then ethanol production capability is improved, but ability to degrade starch and proteins in kitchen waste deteriorates

Engineering Contradiction:
Improveethanol production capabilityVSAvoidability to degrade starch and proteins
Core Design Contradiction:
ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent combines multiple enzyme genes (amylase, glucoamylase, and protease genes from Aspergillus oryzae and Aspergillus niger) into a single multigene co-expression vector, which is then integrated into the Candida utilis genome. This merging of genetic functions enables the yeast to simultaneously produce all necessary enzymes for starch and protein degradation, resolving the contradiction between maintaining ethanol production capability and gaining the ability to utilize kitchen waste components.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent creates a composite genetic system by integrating multiple heterologous genes from different sources (Aspergillus oryzae and Aspergillus niger) into the Candida utilis genome using a multigene co-expression vector. This composite genetic construction enables the yeast to function as a multi-functional organism capable of degrading both starch and proteins while maintaining its ethanol fermentation capability.

Inventive Principle:
Principle #40Composite materials

2Adaptability or versatility

If multiple enzymes are introduced to degrade kitchen waste, then degradation capability is improved, but genetic engineering complexity increases

Engineering Contradiction:
Improvedegradation capabilityVSAvoidgenetic engineering complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent merges multiple enzyme-encoding genes (amylase, glucoamylase, and protease genes) into a single multigene co-expression vector. This consolidation reduces the number of separate genetic manipulation steps required, as all enzymes are introduced simultaneously through one transformation process rather than requiring separate introductions for each enzyme, thereby reducing overall genetic engineering complexity.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The multigene co-expression vector serves as a universal platform that simultaneously delivers multiple enzyme functions. This multi-functional vector design simplifies the genetic engineering process by providing a single tool that achieves what would otherwise require multiple separate genetic constructions and transformations.

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

3Loss of substance

If kitchen waste is used as raw material, then resource utilization is improved, but environmental pollution from traditional treatment methods persists

Engineering Contradiction:
Improveresource utilizationVSAvoidenvironmental pollution
Core Design Contradiction:
Loss of substanceVSObject-generated harmful factors

Solution Approach 1:

The patent converts kitchen waste, which traditionally causes environmental pollution through rotting, landfilling, or incineration, into a beneficial resource for ethanol production. By engineering Candida utilis to directly degrade and ferment kitchen waste components, the process transforms a harmful waste stream into a valuable fuel source, eliminating pollution while achieving resource utilization.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

Solution Approach 2:

The genetically engineered Candida utilis performs self-service by producing all necessary enzymes (amylase, glucoamylase, and protease) to degrade starch and proteins in kitchen waste, then fermenting the resulting sugars to produce ethanol. This self-sufficient system eliminates the need for separate pre-treatment and enzyme addition steps, enabling direct conversion of kitchen waste to ethanol while avoiding the environmental issues associated with traditional treatment methods.

Inventive Principle:
Principle #25Self-service

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 genetically engineered Candida utilis can efficiently convert kitchen waste into ethanol, achieving a high sugar alcohol conversion rate and simplifying industrial application by reducing operational complexity and equipment costs.

Implementation Method 1

Candida utilis lacks enzymes for effectively degrading starch into glucose... it is necessary to introduce the genes of enzymes which are capable of degrading starch and proteins into Candida utilis by genetic engineering

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

Candida utilis lacks enzymes for effectively degrading starch into glucose, and degrading proteins into polypeptides and amino acids

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 3

Candida utilis... can produce ethanol by glucose fermentation and has a comparable sugar alcohol conversion rate to that of Saccharomyces cerevisiae

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentEP3556846B1Genetically engineered candida utilis capable of degrading and utilizing kitchen waste and construction method therefor
Publication Date: 2022.12.07 GUANGDONG RECYCLEAN LOW CARBON TECH CO LTD
  • EP3556846B1 patent drawingFigure 1
  • EP3556846B1 patent drawingFigure 2a~2b
  • EP3556846B1 patent drawingFigure 2c

AI summary

Provided is a genetically engineered Candida utilis capable of degrading and utilizing kitchen waste. The genetically engineered Candida utilis is obtained by using a Candida utilis multigene co-expression vector to integrate alpha-amylase, glucoamylase and acid protease genes into the Candida utilis genome and to correctly express such three enzymes.