NMN Biocatalysis Using Ribose and Enzyme Segmentation
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Solution Overview
Problem
Current methods for preparing nicotinamide mononucleotide (NMN) by biocatalysis are limited by the high cost and scarcity of PRPP, which restricts large-scale industrial production and increases production costs.
Innovation Solution
A method involving catalytic reaction of nicotinamide, ATP, and ribose in the presence of Nampt, ribose phosphate pyrophosphokinase, and ribokinase, avoiding the use of PRPP, with immobilized enzymes and mutant Nampt enzymes to enhance catalytic activity and reduce costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If PRPP is used as substrate in biocatalytic method, then NMN can be prepared, but production cost increases due to high price and limited sources of PRPP
Solution Approach 1:
The patent segments the PRPP-dependent reaction pathway into multiple independent enzymatic steps. Instead of using PRPP as a single substrate, the invention uses ribose as the starting material and employs ribokinase to phosphorylate it to ribose-5-phosphate, then uses ribose phosphate pyrophosphokinase to convert ribose-5-phosphate to PRPP in situ. This segmentation eliminates the need to purchase expensive external PRPP while maintaining the required biochemical transformations.
Solution Approach 2:
The patent implements a self-service system where the enzyme system itself generates the required PRPP substrate from inexpensive ribose. The coupled enzymes ribokinase and ribose phosphate pyrophosphokinase work together to convert ribose to PRPP within the reaction system, making the system self-sufficient and eliminating dependence on external expensive substrates.
2Ease of manufacture
If PRPP is used as substrate, then biocatalytic preparation can proceed, but large-scale industrial production is restricted due to limited sources of PRPP
Solution Approach 1:
By segmenting the pathway into ribose phosphorylation and PRPP generation steps catalyzed by separate enzymes, the invention enables each step to be optimized independently for industrial scale-up. The use of ribose as a commodity chemical with abundant supply chains facilitates large-scale production that would be impossible with limited PRPP sources.
Solution Approach 2:
The patent makes the enzyme system universally applicable to large-scale production by replacing the specialized, limited-substrate PRPP approach with a universal ribose-based pathway. Ribose is a commodity chemical available in large quantities, making this approach universally scalable for industrial production across different applications and scales.
3Ease of manufacture
If conventional biocatalytic method with PRPP is used, then NMN production is achieved, but environmental pollution occurs due to organic solvent residue
Solution Approach 1:
The patent employs inexpensive, easily degradable enzymes that can be used in crude form without requiring organic solvents for purification or stabilization. The enzymes perform their catalytic function and then can be disposed of or degraded naturally, eliminating the need for persistent organic solvents that会造成 environmental pollution.
Solution Approach 2:
The invention changes the reaction parameters from conventional organic-solvent-based biocatalysis to an aqueous-based enzyme system. By optimizing enzyme activity and reaction conditions in aqueous environments, the patent achieves effective NMN production without introducing harmful organic solvents into the process.
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
This method achieves a high conversion rate of up to 100% based on substrate ATP, making it environmentally friendly, cost-effective, and suitable for large-scale industrial production of NMN.
Implementation Method 1
nicotinamide and 5'-phosphoribosyl-1'-pyrophosphate (PRPP) are generally used as substrates for preparing NMN in the presence of nicotinamide phosphoribosyltransferase (Nampt) as a catalyst
Implementation Method 2
ribose phosphate pyrophosphokinase
Implementation Method 3
ribokinase
Data Source
AI summary
The present invention provides a method for preparing nicotinamide mononucleotide (NMN) by bioanalysis. The method includes a step of catalytically reacting a plurality of raw materials including nicotinamide, ATP, and ribose in the presence of nicotinamide phosphoribosyltransferase (Nampt), ribose phosphate pyrophosphokinase, and ribokinase, to prepare the NMN.