Beta-glucosidase Hydrolysis for Rebaudioside M Production

Solution Overview

Problem

The production of steviol glycosides, particularly rebaudioside M, is challenging due to their low abundance in Stevia rebaudiana leaves and the difficulty in isolating individual glycosides, leading to inconsistent taste profiles and high production costs.

Innovation Solution

A method involving the use of beta-glucosidase enzyme to hydrolyze steviol glycosides in recombinant microbial cells, enhancing the production of rebaudioside M by removing glucosyl groups from substrates like stevioside and rebaudioside A, resulting in increased yields and improved taste characteristics.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If extraction processes are used to obtain steviol glycosides from Stevia rebaudiana leaves, then steviol glycosides can be isolated, but the production cost increases and the desired rebaudioside M is extremely difficult to source due to its low abundance

Engineering Contradiction:

Core Design Contradiction:

Quantity of substanceVSEase of manufacture

Solution Approach 1:

The patent applies preliminary action by using recombinant microbial cells to pre-convert stevioside and rebaudioside A into rebaudioside M before extraction. The microbial cells are engineered with specific enzymatic pathways that transform abundant precursors into the desired rare compound, effectively performing the conversion action in advance and making rebaudioside M more available for extraction.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses recombinant microbial cells as intermediary agents to bridge the gap between abundant steviol glycosides (stevioside, rebaudioside A) and the desired rare compound (rebaudioside M). These engineered cells act as biological factories that convert readily available substrates into the target product through controlled enzymatic reactions, facilitating the transformation without requiring direct extraction from plant leaves.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If recombinant microbial cells are used to produce steviol glycosides, then production can be enhanced, but a variety of different steviol glycosides are produced leading to inconsistent taste profiles

Engineering Contradiction:

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent applies local quality by engineering recombinant microbial cells with specific enzymatic pathways that are locally optimized to produce particular steviol glycosides (stevioside, rebaudioside A, and rebaudioside M) in controlled amounts. By carefully selecting and configuring the enzymatic functions within the microbial cells, the process achieves both high productivity and consistent product composition, ensuring reliable taste profiles.

Inventive Principle:

Principle #3Local quality

3Quantity of substance

If natural steviol glycosides are used as sweeteners, then high intensity sweetness is achieved, but the taste profile varies strongly among different batches due to variations in plant production and extraction conditions

Engineering Contradiction:

Core Design Contradiction:

Quantity of substanceVSStability of the object's composition

Solution Approach 1:

The patent applies self-service by using recombinant microbial cells that autonomously convert stevioside and rebaudioside A into rebaudioside M through their engineered enzymatic pathways. This self-contained biological conversion system eliminates dependence on variable plant production conditions and extraction methods, ensuring consistent composition and taste profile while maintaining high sweetness intensity characteristic of natural steviol glycosides.

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

This approach increases the production of rebaudioside M, improves the taste profile of stevia extracts, and reduces production costs by utilizing a more efficient and cost-effective bioconversion process, providing a consistent and desirable sweetener for the food and beverage industry.

Implementation Method 1

beta-glucosidase to hydrolyze specific substrates present in disrupted recombinant cells

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

the use of beta-glucosidase to hydrolyze specific substrates

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentUS11414689B2Hydrolysis of steviol glycosides by beta-glucosidase

Publication Date: 2022.08.16

AI summary

The present disclosure relates to the use of beta-glucosidase to enhance the production efficiency of desired steviol glycosides, such as rebaudioside M (reb M).