TagF-like Capsule Polymerase Synthesizing Dimeric Polysaccharides

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Solution Overview

Problem

Current capsule polymerases can only synthesize polysaccharides with either glycosidic or phosphodiester linkages, but not both, and are limited to producing trimeric repeating units, whereas a capsule polymerase capable of synthesizing dimeric repeating units with both linkage types is needed.

Innovation Solution

The development of TagF-like capsule polymerases that combine glycosyltransferase and hexose-/alditol-phosphate transferase activity, featuring a TagF-like domain with either a GT-A or GT-B fold, enabling the synthesis of polysaccharides with dimeric repeating units through both glycosidic and phosphodiester linkages.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If conventional capsule polymerases are used, then polysaccharides with either glycosidic or phosphodiester linkages can be synthesized, but not both linkage types in the same polymer

Engineering Contradiction:

Improvelinkage type synthesis capabilityVSAvoidfunctional accuracy

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent combines glycosyltransferase and hexose-/alditol-phosphate transferase activities into a single TagF-like capsule polymerase enzyme. This merging of previously separate enzymatic functions into one multifunctional catalyst enables the synthesis of polysaccharides containing both glycosidic and phosphodiester linkages within the same polymer chain, directly resolving the contradiction between versatility and reliability.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The TagF-like capsule polymerase exhibits multi-functionality by performing both glycosyltransferase and hexose-/alditol-phosphate transferase activities. This universal enzyme can synthesize diverse linkage types (both glycosidic and phosphodiester bonds) in a single catalytic system, achieving the desired versatility without compromising functional accuracy through the unified enzyme structure.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Adaptability or versatility

If conventional capsule polymerases are used, then polysaccharides with trimeric repeating units can be synthesized, but not dimeric repeating units

Engineering Contradiction:

Improverepeating unit structure capabilityVSAvoidstructural accuracy

Core Design Contradiction:

Adaptability or versatilityVSManufacturing precision

Solution Approach 1:

The TagF-like capsule polymerase merges domain structures (TagF-like domain with GT-A or GT-B fold) to create a unified enzyme capable of synthesizing dimeric repeating units. This structural merging enables precise control over the repeating unit architecture, achieving both the desired versatility in structure type and the manufacturing precision required for accurate dimeric unit formation.

Inventive Principle:

Principle #5Merging (Combining)

3Adaptability or versatility

If a single polymerase is designed to perform multiple functions, then versatility improves, but enzyme complexity increases

Engineering Contradiction:

Improvefunctional capabilityVSAvoidenzyme structure complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent merges multiple enzymatic functions into the TagF-like capsule polymerase structure, accepting increased structural complexity as the necessary trade-off for achieving the desired functional versatility of synthesizing both linkage types and dimeric repeating units.

Inventive Principle:

Principle #5Merging (Combining)

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

Enables the production of polysaccharides with dimeric repeating units using TagF-like capsule polymerases, expanding the capabilities of capsule polymerases and potentially enhancing their application in vaccine development and bacterial capsule synthesis.

Implementation Method 1

The TagF-like capsule polymerase family of the present invention, a new family of polymerases, combines glycosyltransferase and hexose-/alditol-phosphate transferase activity

Methodology Applied

Scientific EffectGlycosyltransferase activity: Enzyme

Implementation Method 2

The TagF-like capsule polymerase family of the present invention, a new family of polymerases, combines glycosyltransferase and hexose-/alditol-phosphate transferase activity

Methodology Applied

Scientific EffectHexose-/alditol-phosphate transferase activity: Enzyme

Data Source

PatentUS11891598B2Means and methods for producing phosphate containing capsular polysaccharides

Publication Date: 2024.02.06 MEDIZINISCHE HOCHSCHULE HANNOVER

AI summary

The present invention relates to a host cell, which comprises under the control of a heterologous promoter a polynucleotide comprising a nucleotide sequence encoding a polypeptide capable of synthesizing a polysaccharide consisting of a dimeric repeating unit as well as to a vaccine composition comprising such host cell. Furthermore, either such host cell or a polypeptide expressed by such host cell is used for the production of a polysaccharide consisting of a dimeric repeating unit which may be used as a glycoconjugate vaccine.