Polypeptides Enhancing Transgalactosylation at Low Lactose
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Solution Overview
Problem
Current enzymes used in the dairy industry for producing galacto-oligosaccharides require high lactose concentrations and predominantly exhibit beta-galactosylase activity, limiting their efficiency in producing galacto-oligosaccharides at low lactose levels, such as in milk-based products.
Innovation Solution
Development of isolated polypeptides with transgalactosylating activity, having an amino acid sequence with at least 66% sequence identity to specific mature polypeptides, which can efficiently produce galacto-oligosaccharides in situ when incubated with lactose, even at low concentrations, by balancing transgalactosylation and hydrolysis activities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional beta-galactosidase enzymes are used, then lactose hydrolysis activity is achieved, but transgalactosylating activity is insufficient at low lactose concentrations
Solution Approach 1:
The patent applies parameter changes by modifying the enzyme's amino acid sequence to alter its catalytic properties. Specifically, the invention identifies and modifies key amino acid residues in the beta-galactosidase enzyme to shift its activity profile from predominantly hydrolytic to having enhanced transgalactosylating activity at low lactose concentrations, thereby resolving the contradiction between productivity and substrate concentration requirements
Solution Approach 2:
The patent employs inversion by reversing the conventional enzyme behavior. Instead of using enzymes that require high lactose concentrations for transgalactosylation, the invention creates an enzyme that exhibits high transgalactosylating activity at low lactose concentrations, effectively inverting the traditional concentration-activity relationship
2Productivity
If beta-galactosidase enzymes are used, then hydrolysis activity is predominant, but transgalactosylation activity is limited
Solution Approach 1:
The patent modifies specific amino acid parameters in the enzyme sequence to change the catalytic mechanism. By substituting key residues in the catalytic domain, the enzyme's preference shifts from hydrolysis to transgalactosylation, achieving reliable and efficient transgalactosylation activity
3Adaptability or versatility
If high lactose concentrations are used, then transgalactosylase activity is achieved, but applicability to milk-based products is limited
Solution Approach 1:
The patent changes the enzymatic parameters through amino acid substitution to enable function at low substrate concentrations. This allows the enzyme to be adapted for use in milk-based products where lactose concentration is naturally low, resolving the contradiction between activity requirements and product applicability
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
These polypeptides enable high-efficiency production of galacto-oligosaccharides in dairy products, enhancing the growth of health-promoting Bifidobacterium species and providing long-term disease protection benefits, such as colorectal cancer prevention, while reducing lactose levels.
Implementation Method 1
at high lactose concentrations some β -galactosidases are able to transfer galactose to the hydroxyl groups of D-galactose or D-glucose in a process called transgalactosylation whereby galacto-oligosaccharides are produced
Implementation Method 2
The enzyme β-galactosidase (EC 3.2.1.23) usually hydrolyses lactose to the monosaccharides D-glucose and D-galactose
Data Source
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AI summary
The present invention relates to polypeptides, specifically polypeptides having transgalactosylating activity and nucleic acids encoding these, and their uses in e.g. dairy product