Recombinant Globin Polypeptide Production in Yeast for Meat Substitutes

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Solution Overview

Problem

Current methods for producing globin polypeptides, such as leghemoglobin from plants and myoglobin from bovine, are inefficient, particularly for use in food products like meat substitutes, due to challenges in producing abundant amounts of apoproteins and heme, and existing systems lack effective alternatives for plant-derived globins.

Innovation Solution

A method involving transforming host cells like yeast or bacteria with expression vectors encoding globin polypeptides, followed by culturing and recovering the expressed globin polypeptides, which includes using specific promoters and tag proteins for enhanced expression and purification, and incorporating these globins into meat substitute food products for improved aroma, appearance, flavor, and texture.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If E. coli is used to produce human hemoglobin, then production efficiency is improved, but endotoxin removal cost increases significantly

Engineering Contradiction:

Improvehemoglobin production efficiencyVSAvoidendotoxin removal cost

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent uses yeast cells as a disposable production system that naturally lacks endotoxins. Instead of using E. coli which requires expensive endotoxin removal, the invention switches to yeast as a different biological platform that inherently produces cleaner products, eliminating the need for costly purification steps while maintaining high production efficiency

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent changes the biological host parameter from E. coli to yeast, fundamentally altering the production system's characteristics. This parameter change transforms the product purity profile by selecting an organism that does not produce endotoxins, thereby resolving the contradiction between productivity and manufacturing ease

Inventive Principle:

Principle #35Parameter changes

2Ease of manufacture

If yeast strains are used to produce hemoproteins, then endotoxin issues are avoided, but simultaneous production of apoproteins and heme becomes difficult

Engineering Contradiction:

Improveproduct purityVSAvoidhemoprotein production amount

Core Design Contradiction:

Ease of manufactureVSProductivity

Solution Approach 1:

The patent segments the hemoprotein production into two separate expression systems: one for apoprotein production and one for heme production. By using different yeast strains optimized for each component and then combining them, the system overcomes the limitation of simultaneous production while maintaining high purity and yield

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent introduces an intermediary mixing step where apoprotein and heme are produced separately in different yeast cultures and then combined. This intermediary process allows each component to be optimized independently while achieving high overall productivity in the final hemoprotein product

Inventive Principle:

Principle #24Intermediary (Mediator)

3Object-affected harmful factors

If fat content in meat substitute is reduced, then health benefits increase, but sensory attributes like juiciness and mouthfeel deteriorate

Engineering Contradiction:

Improvesaturated fat contentVSAvoidsensory quality

Core Design Contradiction:

Object-affected harmful factorsVSEase of operation

Solution Approach 1:

The patent introduces globin polypeptides as an intermediary substance that mediates between low fat content and high sensory quality. The globins, which have inherent flavor and texture properties, compensate for the reduced fat by providing meat-like sensory attributes through their molecular structure and interaction with other food components

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the compositional parameters of the meat substitute by incorporating recombinant globin polypeptides at specific concentrations. This parameter change allows the formulation to maintain sensory quality without relying on high fat content, as the globins provide alternative pathways for achieving desirable texture and flavor

Inventive Principle:

Principle #35Parameter changes

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 method enables the efficient production of globin polypeptides with desired properties, enhancing the sensory attributes of meat substitute products by providing improved juiciness, mouthfeel, and flavor, addressing the need for more desirable plant-based meat alternatives.

Implementation Method 1

transforming a host cell with an expression vector comprising one or more nucleic acid sequence(s) encoding said globin polypeptide; culturing said host cell under conditions effective for the expression of said one or more nucleic acid sequence(s); expression of said one or more nucleic acid sequence(s)

Methodology Applied

Scientific EffectGene expression:

Data Source

PatentUS20230416764A1Method of producing globin polypeptide recombinantly and meat substitute food product

Publication Date: 2023.12.28 PLANEAT FOODS PTE LTD

AI summary

The present invention refers to a method of producing globin polypeptide recombinantly and a method of producing globin polypeptide with a cell-free translation system. Further, the present invention provides a food product, preferably a meat substitute food product. The present invention is also directed to a vector and a system for recombinant globin polypeptide production as well as to a cell comprising a recombinant expression vector or one or more recombinant nucleic acid molecule(s).