Sequential Enzyme Treatment for Galactooligosaccharide Yield
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Solution Overview
Problem
Existing methods for producing galactooligosaccharides (GOS) from lactose face challenges in maximizing the degree of transgalactosylation while minimizing lactose hydrolysis, leading to reduced yields and an imbalance in the chemical composition and structure of GOS, which affects their properties and effectiveness in promoting gut health.
Innovation Solution
A method involving the sequential use of acid fungal lactase and yeast lactase enzymes, where the initial lactose solution is incubated with acid fungal lactase to reduce lactose concentration and then with yeast lactase to enhance transgalactosylation, adjusting pH and enzyme concentrations to optimize the production of GOS with a desired composition and structure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If reaction conditions favor enzymatic digestion of lactose to less than 20% of initial concentration, then lactose hydrolysis is maximized, but GOS yield is reduced due to concurrent digestion of synthesized GOS
Solution Approach 1:
The patent divides the enzymatic reaction into two distinct stages using different lactase enzymes: first an acid fungal lactase for initial transgalactosylation when lactose concentration is high, then a neutral lactase for final lactose removal when GOS concentration is high. This segmentation allows each enzyme to operate under optimal conditions without the adverse effects that would occur if a single enzyme attempted to achieve both goals simultaneously.
Solution Approach 2:
The acid fungal lactase performs preliminary transgalactosylation action while lactose concentration is still high, building up GOS reserves before the neutral lactase is introduced. This preliminary action ensures that GOS is synthesized early enough to withstand subsequent hydrolysis by the neutral lactase, maximizing final GOS yield.
2Quantity of substance
If a single lactase enzyme is used to maximize lactose hydrolysis, then lactose concentration is reduced effectively, but the degree of transgalactosylation is insufficient leading to reduced GOS yield
Solution Approach 1:
The patent changes the pH parameter from acidic to neutral between the two enzymatic stages. The acid fungal lactase operates at acidic pH (optimal for transgalactosylation), then the pH is adjusted to neutral for the neutral lactase (optimal for lactose hydrolysis). This parameter change allows each enzyme to function at its optimum, resolving the contradiction between transgalactosylation efficiency and lactose hydrolysis completeness.
Solution Approach 2:
The patent uses a composite enzymatic system combining two different lactase enzymes with distinct properties (acid fungal and neutral) rather than relying on a single enzyme. This composite approach leverages the complementary strengths of each enzyme type to achieve both high transgalactosylation and complete lactose hydrolysis.
3Manufacturing precision
If high lactose concentration is maintained to favor transgalactosylation, then GOS composition and structure are improved, but lactose hydrolysis is insufficient
Solution Approach 1:
The patent segments the process into two phases: Phase 1 maintains high lactose concentration for optimal transgalactosylation and GOS structure formation using acid fungal lactase; Phase 2 reduces lactose concentration using neutral lactase after GOS structure is established. This segmentation allows manufacturing precision to be achieved in Phase 1 without compromising complete lactose hydrolysis in Phase 2.
Solution Approach 2:
The patent dynamically adjusts enzyme type and pH conditions throughout the process. Initially, acidic conditions with acid fungal lactase favor transgalactosylation and GOS structure formation. Later, conditions are dynamically changed to neutral pH with neutral lactase to complete lactose hydrolysis. This dynamic adjustment resolves the contradiction between maintaining high lactose concentration for structure quality and reducing it for complete hydrolysis.
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 effectively increases the yield and diversity of GOS, achieving a balanced composition and structure that enhances their health-promoting properties, such as improving mineral absorption and reducing cholesterol levels, while minimizing lactose hydrolysis.
Implementation Method 1
incubating an initial aqueous solution comprising lactose at an initial concentration with an acid fungal lactase to produce an intermediate aqueous solution comprising lactose and GOS
Implementation Method 2
β-D-galactoside galactohydrolases catalyze the hydrolysis of the galactosyl moiety from the non-reducing end of lactose
Implementation Method 3
β-D-galactoside galactohydrolases can catalyze transgalactosylation in which a galactosyl moiety is transferred to a nucleophilic acceptor other than water
Implementation Method 4
adding a yeast lactase to the intermediate aqueous solution; and incubating the intermediate aqueous solution comprising the yeast lactase to produce a final aqueous solution
Implementation Method 5
β-D-galactoside galactohydrolases catalyze the hydrolysis of the galactosyl moiety from the non-reducing end of lactose
Implementation Method 6
β-D-galactoside galactohydrolases can catalyze transgalactosylation in which a galactosyl moiety is transferred to a nucleophilic acceptor other than water
Data Source
AI summary
Methods are disclosed for the enzymatic preparation of galactooligosaccharide (GOS) from lactose using two different microbial lactase enzymes to maximize the extent of transgalactosylation during the digestion of lactose. Methods are also disclosed for avoiding the turbidity of a solution comprising GOS and lactose as it is adjusted for incubation with a yeast neutral lactase.