Lactase Variant Amino Acid Substitutions for Low-Temperature Activity
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Solution Overview
Problem
Current lactase enzymes used in industry are costly due to low activity at low temperatures and inhibition by galactose, requiring high dosages and increasing costs for producing lactose-reduced or lactose-free dairy products.
Innovation Solution
Development of a lactase variant with specific amino acid substitutions, such as at positions 233, 257, 258, 263, 274, 284, 297, 415, 440, 483, 619, 621, 622, 633, 862, or 1004, which enhances specific activity, reduces galactose inhibition, and increases activity in milk at low temperatures, leading to reduced enzyme dosage and production costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current lactase enzymes are used in industrial applications, then lactose hydrolysis can be achieved, but the enzyme requires high dosages due to low activity at low temperatures and inhibition by galactose, increasing production costs
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions at positions 233, 257, 258, 263, 274, 284, 297, 415, 440, 483, 619, 621, 622, 633, 862, or 1004 in the lactase enzyme sequence. These substitutions modify the enzyme's catalytic properties to enhance activity at low temperatures and reduce galactose inhibition, thereby improving productivity while reducing the quantity of enzyme required
Solution Approach 2:
The patent creates a copy of the natural lactase enzyme with modified amino acid sequences. By copying the enzyme structure and introducing specific substitutions, the invention produces a variant that retains lactose hydrolysis function while overcoming the limitations of low temperature activity and galactose inhibition, reducing the dosage needed for industrial applications
2Reliability
If high dosages of lactase are used to overcome low activity at low temperatures, then sufficient hydrolysis activity is achieved, but production costs increase
Solution Approach 1:
The patent modifies the enzyme's amino acid sequence parameters to change its functional properties. The substitutions at specific positions enhance catalytic efficiency and stability at low temperatures, ensuring reliable hydrolysis activity without requiring increased enzyme dosage, thereby reducing production costs while maintaining manufacturing ease
3Reliability
If high dosages of lactase are used to overcome galactose inhibition, then sufficient hydrolysis activity is maintained, but production costs increase
Solution Approach 1:
The patent introduces amino acid substitutions that specifically address galactose inhibition by modifying the enzyme's active site or regulatory regions. These parameter changes reduce the inhibitory effect of galactose on the enzyme, maintaining reliable hydrolysis activity throughout the process without requiring excessive enzyme dosage, thus lowering production costs
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 lactase variant demonstrates improved specific activity, reduced galactose inhibition, and increased activity in milk at low temperatures, resulting in lower enzyme dosages and production costs, enhancing the efficiency and cost-effectiveness of lactose hydrolysis in dairy products.
Implementation Method 1
lactase or beta-galactosidase (E.C: 3.2.1.23) is an enzyme, which catalyzes the hydrolysis of lactose (a disaccharide) into its component monosaccharides glucose and galactose
Data Source
AI summary
The invention relates to a variant polypeptide having lactase activity. The invention also relates to a nucleic acid sequence encoding such a variant polypeptide, to a nucleic acid construct comprising said nucleic acid sequence, to a recombinant expression vector comprising said nucleic acid construct and to a recombinant host cell comprising said expression vector. Further, the invention relates to a method for producing a lactase variant via use of such a host cell. Also, the invention relates to a method of producing a lactase polypeptide variant. The invention further relates to a composition comprising a lactase variant, to use of such a lactase variant or to the use of a lactase variant-containing composition in the preparation of a dairy product, to a process for the production of a dairy product and to the resulting dairy product.