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Culture Condition Optimization and Pilot Scale Production of the M12 Metalloprotease Myroilysin Produced by the Deep-Sea Bacterium Myroides profundi D25

机译:深海细菌深部细菌D25产生的M12金属蛋白酶Mylyisin的培养条件优化和中试规模生产

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摘要

The protease myroilysin is the most abundant protease secreted by marine sedimental bacterium Myroides profundi D25. As a novel elastase of the M12 family, myroilysin has high elastin-degrading activity and strong collagen-swelling ability, suggesting its promising biotechnological potential. Because myroilysin cannot be maturely expressed in Escherichia coli, it is important to be able to improve the production of myroilysin in the wild strain D25. We optimized the culture conditions of strain D25 for protease production by using single factor experiments. Under the optimized conditions, the protease activity of strain D25 reached 1137 ± 53.29 U/mL, i.e., 174% of that before optimization (652 ± 23.78 U/mL). We then conducted small scale fermentations of D25 in a 7.5 L fermentor. The protease activity of strain D25 in small scale fermentations reached 1546.4 ± 82.65 U/mL after parameter optimization. Based on the small scale fermentation results, we further conducted pilot scale fermentations of D25 in a 200 L fermentor, in which the protease production of D25 reached approximately 1100 U/mL. These results indicate that we successfully set up the small and pilot scale fermentation processes of strain D25 for myroilysin production, which should be helpful for the industrial production of myroilysin and the development of its biotechnological potential.
机译:蛋白酶Mylyilysin是海洋沉积细菌深渊Myroides D25分泌的最丰富的蛋白酶。作为M12家族的新型弹性蛋白酶,髓鞘溶素具有高的弹性蛋白降解活性和强大的胶原蛋白溶胀能力,表明其具有广阔的生物技术潜力。由于Myoilysin不能在大肠杆菌中成熟表达,因此重要的是能够提高野生株D25中Myoilysin的产生。我们通过使用单因素实验优化了菌株D25的培养条件以生产蛋白酶。在优化的条件下,菌株D25的蛋白酶活性达到1137±53.29 U / mL,即为优化前(652±23.78 U / mL)的174%。然后,我们在7.5 L发酵罐中进行了D25的小规模发酵。参数优化后,D25菌株在小规模发酵中的蛋白酶活性达到1546.4±82.65 U / mL。基于小规模发酵的结果,我们进一步在200 L发酵罐中进行了D25的中试规模发酵,其中D25的蛋白酶产量达到约1100 U / mL。这些结果表明,我们成功地建立了D25菌株的小型和中试规模的发酵液生产,这将有助于工业化生产Mylyysin和开发其生物技术潜力。

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