Pyp1 Polyol Phosphatase Engineering for Metabolic Inhibition

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for producing polyols and electron-rich biofuels face limitations due to the lack of understanding of metabolism and the absence of suitable polyol phosphate phosphatases, which hinders the optimization of yield and the accumulation of polyol phosphates that inhibit key metabolic enzymes.

Innovation Solution

A recombinant Saccharomyces cerevisiae strain is engineered with a polynucleotide encoding Pyp1, a polyol phosphatase, to dephosphorylate polyol phosphates, thereby preventing their accumulation and enhancing the production of polyols and electron-rich compounds like isobutanol by regulating NADPH levels and bypassing metabolic inhibitors.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If polyol phosphate phosphatase is introduced to dephosphorylate polyol phosphates, then polyol production yield is improved, but the complexity of the metabolic engineering system increases

Engineering Contradiction:

Improvepolyol production yieldVSAvoidmetabolic engineering system complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent introduces polyol phosphate phosphatase as an intermediary enzyme to convert polyol phosphates into polyols. This enzyme acts as a mediator in the metabolic pathway, facilitating the transformation of inhibitory polyol phosphate intermediates into desired polyol products, thereby resolving the contradiction between improving yield and managing system complexity

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs metabolic engineering to alter key parameters of the yeast system, including overexpressing polyol phosphate phosphatase and adjusting NADPH/NADP+ ratio through pathway modifications. These parameter changes enable the system to achieve high polyol yields while managing the complexity through controlled metabolic adjustments

Inventive Principle:

Principle #35Parameter changes

2Productivity

If NADPH regeneration is upregulated to enhance biofuel production, then electron-rich compound yield is improved, but the accumulation of polyol phosphates inhibits key metabolic enzymes

Engineering Contradiction:

Improveelectron-rich compound yieldVSAvoidmetabolic enzyme inhibition

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent converts the harmful accumulation of polyol phosphates (which inhibit metabolic enzymes) into a beneficial process by introducing polyol phosphate phosphatase. This enzyme transforms the inhibitory polyol phosphates into polyols, turning the harmful accumulation problem into a productive pathway that enhances both polyol and electron-rich compound production

Inventive Principle:

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

Solution Approach 2:

The patent implements feedback mechanisms through metabolic engineering to regulate NADPH/NADP+ ratio and polyol phosphate levels. By monitoring and adjusting these metabolic parameters, the system maintains optimal conditions that prevent enzyme inhibition while maximizing electron-rich compound yield

Inventive Principle:

Principle #23Feedback

3Reliability

If polyol phosphate phosphatase activity is increased to reduce polyol phosphate accumulation, then metabolic inhibition is alleviated, but the cost of metabolic engineering increases

Engineering Contradiction:

Improvemetabolic pathway efficiencyVSAvoidmetabolic engineering cost

Core Design Contradiction:

ReliabilityVSEase of manufacture

Solution Approach 1:

The patent employs self-service principles by utilizing the yeast's own metabolic machinery and endogenous polyol phosphate phosphatase activity. Rather than requiring complex external systems, the engineering focuses on optimizing and regulating the cell's intrinsic metabolic pathways, thereby reducing the cost and complexity of metabolic engineering while maintaining high reliability

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 engineered strain effectively reduces polyol phosphate levels, alleviates metabolic inhibition, and increases the yield of polyols and biofuels by maintaining a high NADPH/NADP+ ratio, thereby improving glycolytic and pentose phosphate pathway fluxes.

Implementation Method 1

A recombinant Saccharomyces cerevisiae strain is engineered with a polynucleotide encoding Pyp1, a polyol phosphatase, to dephosphorylate polyol phosphates

Methodology Applied

Scientific EffectDephosphorylation: Hydrolysis

Implementation Method 2

enhancing the production of polyols and electron-rich compounds like isobutanol by regulating NADPH levels and bypassing metabolic inhibitors

Methodology Applied

Scientific EffectRedox reactions: Redox Reactions

Data Source

PatentUS20230416703A1Methods and Materials for Producing Polyols and Electron Rich Compounds

Publication Date: 2023.12.28 THE TRUSTEES OF PRINCETON UNIV

AI summary

Methods and materials for producing polyols are provided comprising recombinant microorganisms expressing a Pyp1 polyol phosphatase. Also provided herein are methods and materials for producing electron rich compounds in recombinant microorganisms lacking the DET1 and/or PHO13 genes.