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Lytic enzyme production optimization using low‐cost substrates and its application in the clarification of xanthan gum culture broth

机译:利用低成本底物优化裂解酶生产及其在黄原胶培养液澄清中的应用

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AbstractLytic enzymes are widely used in industrial biotechnology as they are able to hydrolyze the bacterial cell wall. One application of these enzymes is the clarification of the culture broth for the production of xanthan gum, because of its viability in viscous media and high specificity. The screening process for filamentous fungi producing lytic enzymes, the optimization of production of these enzymes by the selected microorganism, and the optimization of the application of the enzymes produced in the clarification of culture broth are presented in this article. Eleven fungal isolates were tested for their ability to produce enzymes able to increase the transmittance of the culture broth containing cells of Xanthomonas campestris. To optimize the secretion of lytic enzymes by the selected microorganism the following variables were tested: solid substrate, initial pH, incubation temperature, and addition of inducer (gelatin). Thereafter, secretion of the enzymes over time of incubation was assessed. To optimize the clarification process a central composite rotational design was applied in which the pH of the reaction medium, the dilution of the broth, and the reaction temperature were evaluated. The isolate identified as Aspergillus tamarii was selected for increasing the transmittance of the broth from 2.1% to 54.8%. The best conditions for cultivation of this microorganism were: use of coconut husk as solid substrate, with 90% moisture, at 30°C for 20 days. The lytic enzymes produced thereby were able to increase the transmittance of the culture broth from 2.1% to 70.6% at 65°C, without dilution and without pH adjustment.
机译:摘要裂解酶能够水解细菌细胞壁,因此在工业生物技术中被广泛使用。这些酶的一种应用是澄清用于生产黄原胶的培养液,因为它在粘性介质中具有生存力并且具有高特异性。本文介绍了产生丝状真菌的裂解酶的筛选过程,所选微生物对这些酶的生产的优化以及在澄清培养液中产生的酶的应用的优化。测试了11种真菌分离物产生酶的能力,该酶能够增加含有油菜黄单胞菌的细胞的培养液的透射率。为了优化所选微生物的裂解酶分泌,测试了以下变量:固体底物,初始pH,孵育温度和添加诱导剂(明胶)。此后,评估孵育过程中酶的分泌。为了优化澄清过程,采用了中央复合旋转设计,其中评估了反应介质的pH,肉汤的稀释度和反应温度。选择鉴定为塔氏曲霉的分离物以将肉汤的透射率从2.1%增加至54.8%。培养这种微生物的最佳条件是:在30°C下使用椰壳作为具有90%水分的固体底物20天。由此产生的裂解酶能够在65℃下在不稀释和不调节pH的情况下将培养液的透射率从2.1%增加至70.6%。

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