D-Amino Acid Transferases With Thermostable pH-Tolerant Activity
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Solution Overview
Problem
Current enzymes with d-amino-acid transferase activity lack specificity and stability across varying temperatures and pH conditions, limiting their application in industrial and pharmaceutical contexts.
Innovation Solution
Development of enzymes with d-amino-acid transferase activity encoded by nucleic acids with sequences showing at least 90% to 100% identity to SEQ ID NO: 219, which are thermostable and retain activity under acidic and basic conditions, along with methods for producing and using these enzymes in various formulations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current enzymes with d-amino-acid transferase activity are used, then the basic catalytic function is achieved, but the enzymes lack specificity and stability across varying temperatures and pH conditions
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the enzyme to create variants with improved stability. Specifically, the patent describes modifying residues in the active site and substrate binding regions to enhance thermostability and pH stability while maintaining catalytic activity. This involves changing physical and chemical properties of the enzyme structure through targeted amino acid substitutions.
Solution Approach 2:
The patent creates composite enzyme structures by combining stable structural domains with catalytically active regions. The enzyme variants incorporate stable framework residues from thermophilic sources while retaining the catalytic core, creating a composite structure that exhibits both stability and adaptability across varying temperatures and pH conditions.
2Productivity
If enzyme specificity is improved for particular substrates, then catalytic efficiency increases, but the enzyme may lose stability under industrial processing conditions
Solution Approach 1:
The patent applies local quality by making targeted modifications to specific regions of the enzyme while leaving other regions unchanged. The amino acid substitutions are concentrated in the active site and substrate binding regions to maintain or enhance catalytic efficiency, while the overall structural framework remains stable. This localized modification approach allows optimization of catalytic properties without compromising global stability.
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 enzymes demonstrate enhanced stability and activity across a wide range of temperatures and pH levels, enabling their use in diverse industrial and pharmaceutical applications, including food, feed, and beverage production.
Implementation Method 1
Transferases, e.g., transaminases, e.g., d-amino-acid transferases (also referred to as 'd-aminotransferases' or 'D-ATs')... catalyze transamination, catalyze the reaction: D-alanine + 2-oxoglutarate <=> pyruvate + D-glutamate
Implementation Method 2
oxidoreductases, e.g., dehydrogenases, e.g., d-amino-acid dehydrogenases... catalyze an oxidation-reduction reaction, catalyze the removal of hydrogen atoms
Data Source
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AI summary
This invention relates generally to enzymes, polynucleotides encoding the enzymes, the use of such polynucleotides and polypeptides and more specifically to enzymes having transferase activity, e.g., transaminase activity, e.g., d-amino-acid transferase activity, and/or oxidoreductase activity, e.g., dehydrogenase activity, e.g., damino-acid dehydrogenase activity, and/or catalyze the transfer of a chemical group, catalyze transamination, catalyze the reaction: D-alanine + 2-oxoglutarate <=> pyruvate + D-glutamate, and/or catalyze an oxidation-reduction reaction, catalyze the removal of hydrogen atoms, and/or catalyze the reaction: D-amino acid + H2O + acceptor 3 + reduced acceptor. Thus, the invention provides enzymes, compositions, methods for production of pharmaceutical compositions, pharmaceutical intermediates, antibiotics, sweeteners, peptide enzymes, peptide hormones, fuel and fuel additive compositions, foods and food additives, beverage and beverage additives, feeds and feed additives, drugs and drug additives, dietary supplements, textiles, wood, paper, pulp, and detergents comprising the polypeptides or polynucleotides in accordance with the invention.