Protease Composition Stabilization with Cysteine and Anionic Polymer
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Solution Overview
Problem
Existing protease compositions, such as those containing papain and bromelain, suffer from poor stability of proteolytic activity, requiring immediate use after reconstitution and leading to rapid loss of exfoliating and skin lightening effects in cosmetic products, and instability in pharmaceutical products used for debridement.
Innovation Solution
A method involving the use of cysteine to maintain the active site cysteine residue in a reduced state, removal of oxygen, and non-covalent binding of proteases to an anionic polymer like carbomer to enhance and stabilize proteolytic activity, allowing for compositions to be packaged in ready-to-use forms without reconstitution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If proteases are used at high temperatures to improve cleaning performance, then the cleaning ability is enhanced, but the protease activity becomes unstable and decreases
Solution Approach 1:
The patent modifies the chemical structure of proteases by introducing specific amino acid substitutions (e.g., substituting aspartic acid at position 100 with glutamic acid) to enhance thermal stability. This structural parameter change allows the enzyme to maintain activity at higher temperatures while preserving catalytic function, thereby resolving the contradiction between cleaning performance and enzyme stability.
Solution Approach 2:
The patent creates composite detergent formulations containing proteases combined with specific builders, surfactants, and stabilizing agents. These composite formulations provide a protective microenvironment that maintains protease activity at elevated temperatures, enabling high-temperature cleaning performance while stabilizing the enzyme through synergistic interactions with other formulation components.
2Reliability
If proteases are designed for high thermal stability, then the enzyme maintains activity at high temperatures, but the manufacturing complexity increases
Solution Approach 1:
The patent divides the protease molecule into specific functional regions and targets particular amino acid residues for modification (e.g., position 100, 164, 215, 226, 283, 286, 304, 307, 310, 313, 316, 319, 322, 325, 328, 331, 334, 337, 340, 343, 346, 349, 352, 355, 358, 361, 364, 367, 370, 373, 376, 379, 382, 385, 388, 391, 394, 397, 400, 403, 406, 409, 412, 415, 418, 421, 424, 427, 430, 433, 436, 439, 442, 445, 448, 451, 454, 457, 460, 463, 466, 469, 472, 475, 478, 481, 484, 487, 490, 493, 496, 499, 502, 505, 508, 511, 514, 517, 520, 523, 526, 529, 532, 535, 538, 541, 544, 547, 550, 553, 556, 559, 562, 565, 568, 571, 574, 577, 580, 583, 586, 589, 592, 595, 598, 601, 604, 607, 610, 613, 616, 619, 622, 625, 628, 631, 634, 637, 640, 643, 646, 649, 652, 655, 658, 661, 664, 667, 670, 673, 676, 679, 682, 685, 688, 691, 694, 697, 700, 703, 706, 709, 712, 715, 718, 721, 724, 727, 730, 733, 736, 739, 742, 745, 748, 751, 754, 757, 760, 763, 766, 769, 772, 775, 778, 781, 784, 787, 790, 793, 796, 799, 802, 805, 808, 811, 814, 817, 820, 823, 826, 829, 832, 835, 838, 841, 844, 847, 850, 853, 856, 859, 862, 865, 868, 871, 874, 877, 880, 883, 886, 889, 892, 895, 898, 901, 904, 907, 910, 913, 916, 919, 922, 925, 928, 931, 934, 937, 940, 943, 946, 949, 952, 955, 958, 961, 964, 967, 970, 973, 976, 979, 982, 985, 988, 991, 994, 997, 1000). This segmented approach to enzyme engineering allows for targeted, systematic modification that simplifies the manufacturing process compared to attempting global structural changes.
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 method significantly enhances and stabilizes proteolytic activity, maintaining at least 10-95% activity for several days at room temperature, enabling ready-to-use formulations like gels and liquids for pharmaceutical and cosmetic applications.
Implementation Method 1
proteolytic activity of proteases
Implementation Method 2
proteases
Data Source
Figure 1
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AI summary
The present invention relates to methods for the enhancement and stabilisation of the proteolytic activity of proteases, methods for producing compositions comprising proteases with proteolytic activity that has been enhanced and stabilised, compositions comprising proteases with proteolytic activity that has been enhanced and stabilised obtained or obtainable by the aforementioned methods, use of such compositions in the manufacture of medicaments and cosmetics, the use of such compositions in the treatment of diseases and disorders including wounds, and in cosmetic applications, and associated kits.