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

• Published in: Methods and protocols Vol. 7; no. 2; p. 19
• Main Authors: Melchor-Moncada, Jhon Jairo, García-Barco, Alejandra, Zuluaga-Vélez, Augusto, Veloza, Luz Angela, Sepúlveda-Arias, Juan Carlos
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 25.02.2024; MDPI
• Subjects: Anti-inflammatory agents; Bioreactors; Chromatography; Enzymes; Fermentation; Inflammation; Metalloproteinase; Microorganisms; Nitrogen; Nutrients; Optimization; Production processes; Proteins; Proteolysis; Purification; scaling up; Sericulture; Serratia marcescens; serratiopeptidase; Supply and demand; Ultrafiltration; Variables; fermentation; serratiopeptidase; purification; scaling up; Serratia marcescens
• ISSN: 2409-9279; 2409-9279
• DOI: 10.3390/mps7020019

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.