Food and Nutrition Graduate Program (PPGAN), Federal University of the State of Rio de Janeiro (UNIRIO), Rio de Janeiro 22290-240, RJ, Brazil.
Nutrition School, Federal University of the State of Rio de Janeiro (UNIRIO), Rio de Janeiro 22290-240, RJ, Brazil.
Molecules. 2022 Aug 5;27(15):4981. doi: 10.3390/molecules27154981.
A sequential design strategy was applied to optimize the secretion of pectinases by a strain, from Brazilian sugarcane liquor vat, on passion fruit residue flour (PFRF), through solid-state fermentation (SSF). A factorial design was performed to determine the influence variables and two rotational central composite designs were executed. The validated experimental result was of 7.1 U mL using 50% PFRF (/), pH 5, 30 °C for 24 h, under static SSF. Polygalacturonase, pectin methyl esterase, pectin-lyase and pectate-lyase activities were 3.5; 0.08; 3.1 and 0.8 U mL, respectively. Shotgun proteomics analysis of the crude extract enabled the identification of two pectin-lyases, one pectate-lyase and a glucosidase. The crude enzymatic extract maintained at least 80% of its original activity at pH values and temperatures ranging from 2 to 8 and 30 to 80 °C, respectively, over 60 min incubation. Results revealed that PFRF might be a cost-effective and eco-friendly substrate to produce pectinases. Statistical optimization led to fermentation conditions wherein pectin active proteins predominated. To the extent of our knowledge, this is the first study reporting the synthesis of pectate lyase by .
采用序贯设计策略，通过固态发酵（SSF），从巴西甘蔗渣酒罐中优化菌株对西番莲残渣粉（PFRF）分泌果胶酶。进行了析因设计以确定影响变量，并执行了两个旋转中心组合设计。在静态 SSF 下，验证实验结果为 50% PFRF（/）、pH5、30°C 24 h 时为 7.1 U mL。聚半乳糖醛酸酶、果胶甲酯酶、果胶裂解酶和果胶裂解酶活性分别为 3.5、0.08、3.1 和 0.8 U mL。粗提物的鸟枪法蛋白质组学分析鉴定出两种果胶裂解酶、一种果胶裂解酶和一种葡萄糖苷酶。粗酶提取物在 2 至 8 的 pH 值和 30 至 80°C 的温度范围内孵育 60 分钟后，至少保持其原始活性的 80%。结果表明，PFRF 可能是生产果胶酶的一种具有成本效益和环保的廉价底物。统计优化导致发酵条件下以果胶活性蛋白为主。就我们所知，这是首次报道由 产生果胶裂解酶的研究。
