Recombinant Host Cells for D-Lactic Acid Production

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Solution Overview

Problem

Current methods for producing high-purity D-lactic acid through microbial fermentation are inefficient at commercial scales due to low yields, high costs, and byproduct formation, limiting its application in biodegradable plastics and other industries.

Innovation Solution

Development of recombinant cells with heterologous nucleic acids encoding D-lactate dehydrogenase and other proteins, along with genetic disruptions in genes like pyruvate decarboxylase and NAD(P)H dehydrogenase, to enhance D-lactic acid production and reduce byproduct formation, using a biosynthetic process that utilizes renewable carbon sources.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If existing bacterial fermentation methods are used to produce D-lactic acid, then D-lactic acid can be manufactured, but the yields are low and production costs are high

Engineering Contradiction:
ImproveD-lactic acid yieldVSAvoidproduction cost
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent applies parameter changes by modifying fermentation conditions including pH control (maintaining pH 4.0-6.0), temperature optimization (30-37°C), and dissolved oxygen control (20-80% saturation) to maximize D-lactic acid yield while reducing production costs through efficient process parameters

Inventive Principle:
Principle #35Parameter changes

2Manufacturing precision

If existing microbial fermentation methods are used, then D-lactic acid production is possible, but byproduct formation occurs and purity is reduced

Engineering Contradiction:
ImproveD-lactic acid purityVSAvoidbyproduct formation
Core Design Contradiction:
Manufacturing precisionVSObject-generated harmful factors

Solution Approach 1:

The patent applies the extraction principle by removing harmful byproducts through specific purification steps including centrifugation, filtration, and ion exchange chromatography to isolate high-purity D-lactic acid from the fermentation broth, eliminating contaminants and byproducts

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the harmful effect of byproduct formation into a benefit by using the byproducts as indicators to optimize fermentation conditions, and by utilizing waste heat from the fermentation process for preheating incoming media, thereby improving overall process efficiency

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

3Productivity

If complex and expensive nutrients are used in fermentation media, then microbial growth is supported, but production costs increase and commercial scale application is prohibited

Engineering Contradiction:
Improvefermentation efficiencyVSAvoidmedia cost
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent applies this principle by using inexpensive, readily available nutrients in the fermentation media such as yeast extract, peptone, and glucose, replacing complex and expensive defined media components while maintaining high fermentation efficiency and enabling commercial scale production

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

4Manufacturing precision

If engineered host cells are used in Saccharomyces cerevisiae, then D-lactic acid production is attempted, but the cells cannot grow leading to low yields

Engineering Contradiction:
ImproveD-lactic acid yieldVSAvoidhost cell growth
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The patent resolves the contradiction by optimizing fermentation parameters including maintaining pH 4.0-6.0, temperature 30-37°C, and dissolved oxygen 20-80% saturation, which allows engineered S. cerevisiae cells to both grow and produce D-lactic acid at high yields

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies dynamics by using a two-stage fermentation process where the first stage optimizes for cell growth and the second stage optimizes for D-lactic acid production, allowing the system to adapt to different operational requirements and achieve both high cell density and high product yield

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

The approach enables high-yield, high-purity D-lactic acid production with reduced byproduct formation, making it suitable for commercial applications in biodegradable plastics and other industries, and aligns with the need for sustainable, renewable chemical alternatives to petrochemicals.

Implementation Method 1

recombinant cells with heterologous nucleic acids encoding D-lactate dehydrogenase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

production of D-lactic acid by direct fermentation from carbon sources

Methodology Applied
Scientific EffectFermentation: Fermentation

Data Source

PatentUS11390891B2Recombinant host cells and methods for the production of D-lactic acid
Publication Date: 2022.07.19 LYGOS INC

AI summary

Methods and materials related to producing D-lactic acid are disclosed. Specifically, isolated synthetic or natural nucleic acids, synthetic or natural polypeptides, host cells, and methods and materials for producing D-lactic acid by direct fermentation from carbon sources are disclosed, along with methods of preparing D-lactic acid polymers.