两株链球菌在低成本培养基中产链激酶的体外检测与优化

M. El-Mongy, T. Taha
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引用次数: 3

摘要

本研究旨在证明链激酶生产的优化。对化脓性链球菌和同种链球菌在不同培养基中生长过程中的产酶情况进行了监测。当这两种微生物在链球菌基培养基上生长时,在孵育过程中每12小时调整一次化脓性链球菌和同种链球菌培养基的pH值,可以显著提高酶的产量。产链激酶的最佳碳源是葡萄糖,而甘露醇和山梨醇是不合适的碳源。当微生物在Strep-base培养基上生长时,酵母提取物和酪蛋白可作为链激酶生产有机氮的主要来源。当色氨酸和酪蛋白含量分别为1.5% (w/v)和1.5% (w/v)时,产酶量最高。采用普通酪蛋白消化法和更灵敏的酶底物消化法检测链激酶。此外,使用蛋白蛋白敏感电极对酶进行电化学检测,以比较不同的检测方法。电化学方法得到的结果与其他方法得到的结果非常接近。这些结果为微生物生长过程中链激酶的检测提供了可靠的替代方法。它为链激酶的测定提供了一种更快、更便宜的技术,特别是当需要在浑浊介质中检测酶时。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
In vitro detection and optimization of streptokinase production by two streptococcal strains in a relatively low cost growth medium
This study sought to demonstrate the optimization of Streptokinase Production. Enzyme production was monitored during the growth of both Streptococcus pyogenes and Streptococcus equisimilis in different media. Adjustment of the pH for culture media of S. pyogenes and S. equisimilis, every 12 hours during incubation, significantly increased the enzyme production levels, when both microbes were grown on Strep-base medium. The best carbon source for streptokinase production was glucose of both S. pyogenes and S. equisimilis, while mannitol and sorbitol were found to be improper carbon sources. Yeast extract, and casein could be used as the primary source of organic nitrogen for streptokinase production, when the microbes were allowed to grow on Strep-base medium. The highest levels of the enzyme production were obtained with 1.5 % (w/v) tryptone and 1.5 % (w/v) casein for S. equisimilis and S. pyogenes, respectively. Detection of streptokinase produced was by the common casein digestion method and by the more sensitive chromozym substrate digestion method. Moreover, the enzyme was assayed electrochemically using the protamine-sensitive electrode to compare different methods of detection. Results obtained from electrochemical method were very close to that obtained with other methods. These results offer alternative and reliable method for streptokinase detection during microbial growth. It provides a faster and less expensive technique for streptokinase determination especially when there is a need to detect the enzyme in turbid media.
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