ABSTRACT
The cross-linked enzyme aggregates (CLEAs) have numerous economic advantages in the industrial bio catalysis. In the present study, the multi CLEAs containing protease, catalase, and lipase from the sunflower seeds using starch as a cofeeder as well as bovine serum albumin (BSA) are designed and prepared successfully. After optimization, multi CLEAs of enzyme have been prepared with ammonium sulfate (55% w/v), glutaraldehyde (100 mM), and 8 mg/mL of starch or 20 mg/mL of BSA. The activity recovery of protease, catalase, and lipase multi CLEAs-starch are 87, 61, and 60%, respectively. Whereas, CLEAs prepared with BSA are 74, 61, and 50% activity and multi CLEAs only 60, 44, and 41% of protease, catalase, and lipase, respectively. The multi CLEAs were used to catalyze the reactions for enhanced washing process. After adding multi CLEAs-starch, the stain removal percentage of detergents is enhanced by 83%.The present study reports a high stability, simplicity, low cost, and recyclability of the novel multi CLEAs from the sunflower seeds that make them efficient as a highly active biocatalysts in the biotechnological applications. We believe that these novel multi CLEAs present a new approach to the synthesis of multi enzyme biocatalysts from the cheap and friendly environmental sources.
Subject(s)
Catalase/chemistry , Helianthus/chemistry , Lipase/chemistry , Peptide Hydrolases/chemistry , Plant Proteins/chemistry , Seeds/chemistry , Ammonium Sulfate/chemistry , Biocatalysis , Catalase/isolation & purification , Coloring Agents/isolation & purification , Cross-Linking Reagents/chemistry , Detergents/chemistry , Enzyme Assays , Glutaral/chemistry , Helianthus/enzymology , Kinetics , Lipase/isolation & purification , Peptide Hydrolases/isolation & purification , Plant Proteins/isolation & purification , Protein Aggregates , Seeds/enzymology , Serum Albumin, Bovine/chemistry , Starch/chemistryABSTRACT
The cross-linked enzyme aggregates (CLEAs) involves formation of a number of covalent bonds between enzyme and the matrix using glutaraldehyde. In general, amino groups of lysine, sulfhydryl groups of cysteine, phenolic OH groups of tyrosine, or imidazol group of histidine are used for enzyme binding under mild conditions. The main advantage of this method is its simplicity, economic advantages in the industrial bio catalysis. The Fe3O4 magnetic nanoparticles were synthesized by coprecipitating Fe2+and Fe3+in alkaline solution. Tannic acid was used to functionalize the Fe3O4 magnetic nanoparticles. After functionalization process, tannic acid magnetic cross-linked enzyme aggregates of enzyme (TA-MNPs-CLEAs) were prepared by cross-linking of enzyme aggregates with different saccharides as additive. The present result reported high stability, simplicity, low cost and recyclability of a saccharide-TA-MNPs-CLEAs-enzyme make it efficient as a highly active biocatalyst in biotechnological applications. The obtained results suggest that disaccharides (maltose, sucrose and lactose) and polysaccharide such as starch are eco-friendly additives to TA-MNPs-lipase and TA-MNPs-CLEAs-peroxidase and can become a powerful biocatalyst in industry applications.
