Scale-Up of the Fermentation Process for the Production and Purification of Serratiopeptidase Using Silkworm Pupae as a Substrate.

Serratiopeptidase, a bacterial metalloprotease known for its pain-relieving and anti-inflammatory properties, can be produced through fermentation with S. marcescens. This study aimed to identify key factors related to nutrient composition and physicochemical conditions for production in Erlenmeyer flasks and to scale up the mixture to a bioreactor to obtain the maximum proteolytic activity. A Plackett-Burman design was used to determine whether the presence of silkworm pupae (at 1.5%) was a significant parameter for serratiopeptidase production. Along with the variables pH, temperature, and time, they were optimized using a Taguchi experimental design, resulting in values of 7, 25 °C, and 36 h, respectively. Scaling up with a kLa of 25.45 ± 3.12 h-1 showed the highest serratiopeptidase production at 24 h. A factorial design was used for ultrafiltration, resulting in an LMH (liters per square meter per hour) of 960 L/m2h, a TMP (transmembrane pressure) of 15 psi, and a concentration factor of five, with a specific activity of 24,325.81 ± 1515.69 U/mg. Afterward, the retentate was purified using strong anion exchange chromatography and ultrafiltration, yielding a 19.94 ± 3.07% recovery and a purification factor of 1.59 ± 0.31. In conclusion, waste from the sericulture industry can be used for serratiopeptidase production.

Efficacy of serratiopeptidase in third molar surgery: a systematic review and meta-analysis / Eficacia de la serratiopeptidasa en cirugía del tercer molar: una revisión sistemática y un metanálisis

Objective: To determine the efficacy of serratiopeptidase in third molar surgery. Materials and Methods: A bibliographic search was carried out until April 2022, in the biomedical databases: Pubmed/Medline, Cochrane Central Registry of Clinical Trials, Scopus, Scielo and Google Scholar. Studies reporting the ef-ficacy of serratiopeptidase in third molar surgery, which were randomized clinical trials, in English and without time limits, were included. The RoB 2.0 tool was used to assess the risk of the included studies and the GRADEPro GDT tool to assess Results: The preliminary search yielded a total of 116 articles, discarding those that did not meet the selection criteria, leaving only 10 articles. Six articles entered a meta-analysis and found that serratiopeptidase reduces trismus but not reduce inflammation and pain after third molar surgery. Conclusions: The literature reviewed suggests that ser-ratiopeptidase is effective in reducing trismus after third molar surgery.

Objetivo: Determinar la eficacia de la serratiopeptidasa en la cirugía del tercer molar. Materiales y Métodos: Se realizó una búsqueda bibliográfica hasta abril de 2022, en las bases de datos biomédicas: Pubmed/Medline, Registro Cochrane Central de Ensayos Clínicos, Scopus, Scielo y Google Scholar. Se incluyeron estudios que reportaron la eficacia de la serratiopeptidasa en cirugía de terceros molares, que fueron ensayos clínicos aleatorios, en inglés y sin límite de tiempo. Se utilizó la herramienta RoB 2.0 para evaluar el riesgo de los estudios incluidos y la herramienta GRADEPro GDT para evaluar la calidad de la evidencia y la fuerza de recomendación de los resultados. Resultados: La búsqueda preliminar arrojó un total de 116 artículos, descartando aquellos que no cumplieron con los criterios de selección, quedando solo 10 artículos. Seis artículos participaron en un metanálisis y encontraron que la serratiopeptidasa reduce el trismo, pero no reduce la inflamación y el dolor después de la cirugía del tercer molar. Conclusión: La literatura revisada sugiere que la serratiopeptidasa es efectiva para reducir el trismo después de la cirugía del tercer molar.

Purification and characterization of a metalloprotease produced by the C8 isolate of Serratia marcescens using silkworm pupae or casein as a protein source.

Serratiopeptidase, a metalloprotease produced by Serratia marcescens, is produced through a fermentation process using carbohydrates and proteins as carbon and nitrogen sources. However, some byproducts of the silk industry could be an alternative source for serratiopeptidase production. Therefore, the present work is focused on the purification and characterization of a serratiopeptidase produced from the C8 isolate of Serratia marcescens and obtained from a Colombian silkworm hybrid using casein or silkworm pupae. The protease was purified using ultrafiltration, anion-exchange, and size-exclusion chromatography. The purified enzyme showed a molecular weight of ~50 kDa with a purity above 96%, an isoelectric point of ~4.6, optimum pH and temperature of 6 and 50 °C, and stability at 4 °C for one month. The kinetic constants using azocasein as substrate were 0.63 mM (Km), 2,016 µM/min (Vmax), 41.41 s-1 (Kcat), and 6.56 × 107 M-1 s-1 (Kcat/Km). Inhibition by 5 mM EDTA or 1,10-phenanthroline was recovered by adding Zn2+ at the same concentration. Mass spectrometry analysis indicated 94% homology with the sequence of serratiopeptidase produced by the E-15 strain. We purified and characterized a serratiopeptidase produced by the C8 isolate of S. marcescens in a culture medium based on a renewable source from the silk industry.