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

IF 2.3 Q3 BIOCHEMICAL RESEARCH METHODS Methods and Protocols Pub Date : 2024-02-25 DOI:10.3390/mps7020019
Jhon Jairo Melchor-Moncada, Alejandra García-Barco, Augusto Zuluaga-Vélez, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias
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Abstract

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.

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利用蚕蛹作为底物生产和纯化塞拉提肽酶的发酵工艺的规模化。
塞拉提肽酶是一种细菌金属蛋白酶,因其具有止痛和消炎的特性而闻名。本研究旨在确定与营养成分和理化条件有关的关键因素,以便在埃尔伦迈耶烧瓶中进行生产,并将混合物放大到生物反应器中,以获得最大的蛋白水解活性。为了确定蚕蛹的存在(1.5%)对丝氨酸肽酶的生产是否是一个重要参数,采用了 "普拉克特-伯曼设计 "法。采用田口试验设计法对 pH 值、温度和时间等变量进行了优化,优化值分别为 7、25 °C 和 36 小时。采用因子设计进行超滤,结果是 LMH(升/平方米/小时)为 960 L/m2h,TMP(跨膜压力)为 15 psi,浓缩因子为 5,比活度为 24,325.81 ± 1515.69 U/mg 。之后,使用强阴离子交换色谱和超滤对回流液进行纯化,回收率为 19.94 ± 3.07%,纯化系数为 1.59 ± 0.31。总之,养蚕业的废弃物可用于生产血清肽酶。
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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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