Nampt Mutant Enzyme Catalytic Activity for NMN Production

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Solution Overview

Problem

The low catalytic activity of wild-type Nampt limits the efficiency and cost-effectiveness of biocatalytic production of nicotinamide mononucleotide (NMN), restricting its industrial application.

Innovation Solution

A Nampt mutant with enhanced catalytic activity is created through site-directed mutagenesis, specifically at positions 180, 182, 231, 298, 338, and 377, which is then expressed using a suitable vector and screened for high activity, allowing for efficient conversion of nicotinamide and PRPP into NMN.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If wild-type Nampt is used for biocatalytic production of NMN, then the process is environmentally friendly and pollution-free, but the catalytic activity is low resulting in low yield and high production cost

Engineering Contradiction:
Improveyield of NMNVSAvoidproduction cost
Core Design Contradiction:
ProductivityVSLoss of energy

Solution Approach 1:

The patent applies parameter changes by modifying the amino acid sequence of Nampt through site-directed mutagenesis. Specific amino acid residues are mutated to alter the enzyme's catalytic properties, thereby increasing its activity and productivity in NMN production while maintaining the environmentally friendly biocatalytic process

Inventive Principle:
Principle #35Parameter changes

2Productivity

If wild-type Nampt is used for biocatalytic production of NMN, then the method is environmentally friendly, but the catalytic activity is low making the product less competitive in the market

Engineering Contradiction:
Improvecatalytic activityVSAvoidindustrial application value
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent modifies the enzyme's parameters through amino acid substitution to enhance catalytic activity. This makes the biocatalytic process more efficient and economically viable for industrial application, thereby improving the ease of manufacture and market competitiveness of NMN

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

The Nampt mutant exhibits 1.2-6.9 times higher enzymatic activity than wild-type Nampt, enabling cost-effective large-scale industrial production of NMN without the need for extensive purification, thereby increasing market competitiveness.

Implementation Method 1

Nampt functions to have an enzyme activity, by which nicotinamide and 5'-phosphoribosyl-1'-pyrophosphate (PRPP) are catalyzed to convert into nicotinamide mononucleotide (NMN) and pyrophosphate

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentUS10174298B2Nicotinamide phosphoribosyltransferase (NAMPT) mutant and use thereof
Publication Date: 2019.01.08 BONTAC BIO ENG (SHENZHEN) CO LTD

AI summary

The present invention discloses a Nicotinamide phosphoribosyltransferase (nampt) mutant and use thereof. The present invention relates to a nicotinamide phosphoribosyltransferase (Nampt) mutant artificially obtained through genic site-directed mutation. An object of the present invention is to provide a Nampt mutant having a catalytic activity higher than that of a conventional wild type parent, wherein the enzymatic activity of the Nampt mutant provided in the present invention is 1.2-6.9 times of the enzymatic activity of the parent.