Bacterial Microcompartments Shield Toxic Protein Expression

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Solution Overview

Problem

The production and purification of non-native, cytotoxic proteins in bacterial systems are challenging due to their toxicity, which can damage cellular components and impair cell function, leading to reduced growth and potential cell death.

Innovation Solution

The method involves introducing polynucleotides encoding microcompartment proteins and toxic non-native proteins into bacterial cells, where the toxic proteins are directed to form empty microcompartments, shielding the cell from toxicity and allowing for the compartmentalization and purification of these proteins.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If toxic non-native proteins are expressed in bacterial cells, then protein production is achieved, but cell growth and function are impaired due to toxicity

Engineering Contradiction:

Improveprotein productionVSAvoidcell growth

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention divides the bacterial cell into functional compartments by introducing microcompartment proteins that self-assemble into discrete structures. These microcompartments segment the cytoplasm to create isolated zones where toxic proteins can be concentrated or sequestered, preventing their harmful effects on the rest of the cell while maintaining overall cell viability and function.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention introduces microcompartment proteins as intermediary structures that act as physical barriers between toxic non-native proteins and native cellular components. These intermediary microcompartments mediate the interaction by providing a protective interface, allowing toxic proteins to be contained within defined spaces without directly damaging cellular membranes or structures.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If microcompartment proteins are introduced to shield toxicity, then cell viability is maintained, but system complexity increases

Engineering Contradiction:

Improvecell viabilityVSAvoidsystem complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The microcompartment proteins introduced into the bacterial system are capable of self-assembly, forming functional microcompartments autonomously without requiring complex external assembly machinery or additional regulatory components. This self-service capability reduces the overall system complexity while maintaining the protective function against toxic proteins.

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The invention utilizes changes in physical parameters such as protein concentration, pH, or ionic strength to trigger the self-assembly of microcompartment proteins into functional structures. By controlling these parameters, the system can dynamically form or dissolve microcompartments as needed, providing a simple regulatory mechanism without requiring complex genetic or biochemical control systems.

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS10155951B2Methods and systems for engineering bacterial systems to shield toxicity during non-native protein expression and purification

Publication Date: 2018.12.18 LAWRENCE LIVERMORE NAT SECURITY LLC

AI summary

The invention described herein relates to methods, systems, compositions and cells to provide one or more toxic non-native proteins in a cell, wherein the one or more toxic non-native proteins are contained in at least one empty microcompartment within the cell.