S53 Protease Variants for Acid-Stable Animal Feed
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Solution Overview
Problem
Current proteases used in animal feed are not efficiently digested in the acidic stomach environment of mono-gastric animals and poultry, and they do not maintain activity after steam treatment in pelleted feed, leading to suboptimal protein digestibility and increased costs due to low yields.
Innovation Solution
Development of isolated polypeptides with at least 84% sequence identity to specific S53 proteases, which are highly acid-stable and maintain activity across a broad pH range, including after steam treatment, for use in animal feed to enhance protein digestibility.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current proteases are used in animal feed, then they can provide protease activity, but they are not efficiently digested in the acidic stomach environment and do not maintain activity after steam treatment
Solution Approach 1:
The patent applies parameter changes by identifying and modifying specific amino acid residues in the protease sequence (e.g., positions 85, 89, 175, 283) to alter the enzyme's pH stability profile. The S53 protease variants are engineered to maintain activity across a broader pH range (pH 2-12) compared to conventional proteases, enabling them to withstand acidic stomach environments and steam treatment while retaining proteolytic function.
2Productivity
If conventional proteases are used in mono-gastric animals, then they can digest proteins, but the apparent ileal protein digestibility is only around 80%
Solution Approach 1:
The patent modifies the protease's functional parameters through amino acid substitutions that enhance acid stability. The S53 protease variants maintain catalytic activity at low pH levels (pH 2-4) typical of the monogastric stomach, thereby improving protein digestibility beyond the conventional 80% threshold by ensuring the enzyme survives and remains active in the acidic gastric environment.
3Ease of manufacture
If proteases are exposed to steam treatment in pelleted feed, then feed processing is completed, but the proteases lose activity
Solution Approach 1:
The patent engineers proteases with enhanced thermal and pH stability parameters through specific amino acid modifications. The S53 protease variants can withstand the high temperature and pH fluctuations of steam treatment during feed pelleting while maintaining proteolytic activity, thereby resolving the contradiction between feed processing requirements and enzyme stability.
4Reliability
If proteases with high acid stability are developed, then they can survive gastric environment, but production costs increase due to lower yields
Solution Approach 1:
The patent optimizes the balance between stability and productivity by making targeted amino acid substitutions that confer acid stability without significantly impacting expression levels. The S53 protease variants achieve enhanced gastric survival while maintaining feasible production yields through rational protein engineering rather than extensive purification or stabilization protocols.
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 S53 proteases effectively digest proteins in the acidic stomach and neutral intestinal environments of animals, improving protein digestibility and reducing production costs through higher yields and stability during feed processing.
Implementation Method 1
proteases which are highly acid stable and so can survive in the gastric environment and at the same time are efficiently active at the broad range of physiological pH of the digestive tract
Implementation Method 2
S53 proteases effectively digest proteins in the acidic stomach and neutral intestinal environments of animals
Data Source
AI summary
The present invention relates to isolated polypeptides having protease activity and isolated nucleic acid sequences encoding the proteases. The invention also relates to nucleic acid constructs, vectors, and host cells, including plant and animal cells, comprising the nucleic acid sequences, as well as methods for producing and using the proteases, in particular the use of the proteases in animal feed.