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
. 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 k
a of 25.45 ± 3.12 h
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/m
h, 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.