ABSTRACT
In this study, horseradish peroxidase was extracted, purified, and immobilized on a calcium alginate-starch hybrid support by covalent bonding and entrapment. The immobilized HRP was used for the biodegradation of phenol red dye. A 3.74-fold purification was observed after precipitation with ammonium sulfate and dialysis. An immobilization yield of 88.33%, efficiency of 56.89%, and activity recovery of 50.26% were found. The optimum pH and temperature values for immobilized and free HRP were 5.0 and 50 °C and 6.5 and 60 °C, respectively. The immobilized HRP showed better thermal stability than its free form, resulting in a considerable increase in half-life time (t1/2) and deactivation energy (Ed). The immobilized HRP maintained 93.71% of its initial activity after 45 days of storage at 4 °C. Regarding the biodegradation of phenol red, immobilized HRP resulted in 63.57% degradation after 90 min. After 10 cycles of reuse, the immobilized HRP was able to maintain 43.06% of its initial biodegradative capacity and 42.36% of its enzymatic activity. At the end of 15 application cycles, a biodegradation rate of 8.34% was observed. In conclusion, the results demonstrate that the immobilized HRP is a promising option for use as an industrial biocatalyst in various biotechnological applications.
ABSTRACT
In this study, horseradish peroxidase (HRP) was immobilized for the first time on Ca alginate-starch hybrid beads and employed for the biodegradation of phenol red dye. The optimal protein loading was 50 mg/g of support. Immobilized HRP demonstrated improved thermal stability and maximum catalytic activity at 50 °C and pH 6.0, with an increase in half-life (t1/2) and enzymatic deactivation energy (Ed) compared to free HRP. After 30 days of storage at 4 °C, immobilized HRP retained 109% of its initial activity. Compared to free HRP, the immobilized enzyme exhibited higher potential for phenol red dye degradation, as evidenced by the removal of 55.87% of initial phenol red after 90 min, which was 11.5 times greater than free HRP. In sequential batch reactions, the immobilized HRP demonstrated good potential efficiency for the biodegradation of phenol red dye. The immobilized HRP was used for a total of 15 cycles, degrading 18.99% after 10 cycles and 11.69% after 15 cycles, with a residual enzymatic activity of 19.40% and 12.34%, respectively. Overall, the results suggest that HRP immobilized on Ca alginate-starch hybrid supports shows promise as a biocatalyst for industrial and biotechnological applications, particularly for the biodegradation of recalcitrant compounds such as phenol red dye.
Subject(s)
Phenol , Phenolsulfonphthalein , Horseradish Peroxidase/chemistry , Alginates/chemistry , Phenols , Enzymes, Immobilized/chemistry , Enzyme StabilityABSTRACT
Microglia are glial cells centrally related to pathophysiology and neuroimmunological regulation of pain through microglia-neuron crosstalk mechanisms. In contrast, anti-inflammatory mechanisms guided by immunological effectors such as IL-10 trigger the secretion of analgesic substances, culminating in the differential expression of genes encoding endogenous opioid peptides, especially ß-endorphin. Thus, when ß-endorphin binds to the µ-opioid receptor, it generates neuronal hyperpolarization, inhibiting nociceptive stimuli. This review aimed to summarize the recent advances in understanding the mechanism by which IL-10/ß-endorphin can reduce pain. For this, databases were searched for articles from their inception up until November 2022. Two independent reviewers extracted the data and assessed the methodological quality of the included studies, and seventeen studies were considered eligible for this review. Several studies have demonstrated the impact of IL-10/ß-endorphin in reducing pain, where IL-10 can stimulate GLP-1R, GRP40, and α7nAChR receptors, as well as intracellular signaling pathways, such as STAT3, resulting in increased ß-endorphin expression and secretion. In addition, molecules such as gabapentinoids, thalidomide, cynandione A, morroniside, lemairamin, and cinobufagin, as well as non-pharmacological treatments such as electroacupuncture, reduce pain through IL-10 mediated mechanisms, reflecting a microglia-dependent ß-endorphin differential increase. This process represents a cornerstone in pain neuroimmunology knowledge, and the results obtained by different studies about the theme are presented in this review.
