优化链霉菌生产果胶酶的培养条件及部分纯化酶的特性。

IF 0.1 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Microbial Physiology Pub Date : 2023-01-01 Epub Date: 2022-11-23 DOI:10.1159/000528257
Sarita Shrestha, Chonlong Chio, Janak Raj Khatiwada, Aristide Laurel Mokale Kognou, Xuantong Chen, Wensheng Qin
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引用次数: 0

摘要

采用 Box-Behnken 设计优化了链霉菌生产果胶酶的培养参数。在含酵母提取物的培养基中进行浸没式发酵,在 35℃、pH 值为 7 的条件下发酵 58 小时后,果胶酶产量最大。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和酶图分析发现,链霉菌产生了两种分子量分别约为 25 和 75 kDa 的果胶酶蛋白。果胶酶活性在很宽的温度范围(30°C-80°C)和 pH 值范围(3-9)内都能检测到,在 70°C 和 pH 值为 5 和 9 时,果胶酶活性最高。即使将酶在不同温度下培养 120 分钟,其活性仍保持在 30-40% 左右。在含有 1.5%果胶、1%酪蛋白作为氮源、0.5 mM MgSO4 和 5 mM NaCl 的培养基中,链霉菌的果胶酶活性增强。此外,在培养基中添加吐温-20、氨基酸和维生素也能提高果胶酶的活性。此外,该细菌还具有高效脱色水晶紫染料的能力。脱色率从 39.29% 到 53.75%不等,在含有 2 mg/mL 结晶紫的培养基中,细菌在 144 小时内的脱色率最高。
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Optimization of Cultural Conditions for Pectinase Production by Streptomyces sp. and Characterization of Partially Purified Enzymes.

The cultural parameters of Streptomyces sp. for pectinase production were optimized using the Box-Behnken design. The maximum pectinase production was obtained after 58 h at 35°C and pH 7 upon submerged fermentation in yeast extract-containing media. The enzymes were partially purified with acetone precipitation, and the analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram revealed that Streptomyces sp. produced two pectinases protein with molecular weights of about 25 and 75 kDa. The pectinase activity was detected in a wide range of temperatures (30°C-80°C) and pH (3-9) with maximum pectinase activities observed at 70°C and pH 5 and 9. The enzymes retained about 30-40% of their activities even after incubating the enzyme at different temperatures for 120 min. The pectinase activities of Streptomyces sp. were enhanced in the media containing 1.5% pectin, 1% casein as a nitrogen source, 0.5 mM MgSO4, and 5 mM NaCl. Further, the addition of Tween-20, amino acids, and vitamins to the media also enhanced the pectinase activity. Moreover, the bacterium illustrated the ability to decolorize crystal violet dye efficiently. The decolorization rate ranged from 39.29 to 53.75%, showing the highest bacterial decolorization in the media containing 2 mg/mL crystal violet at 144 h. Therefore, the bacterium has the potential in treating wastewater produced by industries like textile industries.

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