摘要
A metagenomic library recombinant clone CAPL3, an Escherichia coli strain generated by transformed with metagenomic library from deep-sea sediments, can efficiently produce cold active lipase. The effects of both temperature and dissolved oxygen(DO) on cold active lipase production by batch culture of metagenomic library recombinant clone(CAPL3) from deep-sea sediment were investigated. First, a two-stage temperature control strategy was developed, in which the temperature was kept at 34 ℃ for the first 15 h, and then switched to30 ℃. The cold active lipase activity and productivity reached 315.2 U·ml^-1and 8.08 U·ml^-1·h^-1, respectively,increased by both 14.5% compared to the results obtained with temperature controlled at 30℃. In addition, different DO control modes were conducted, based on the data obtained from the different DO control strategies and analysis of kinetics parameters at different DO levels. A step-wise temperature and DO control strategy were developed to improve lipase production, i.e., temperature and DO level were controlled at 34℃, 30% during 0–15 h;30 ℃, 30% during 15–18 h, and 30 ℃, 20% during 18–39 h. With this strategy, the maximum lipase activity reached 354.6 U·ml^-1at 39 h, which was 28.8% higher than that achieved without temperature and DO control(275.3 U·ml^-1).
A metagenomic library recombinant clone CAPL3, an Escherichia coli strain generated by transformed with metagenomic library from deep-sea sediments, can efficiently produce cold active lipase. The effects of both temperature and dissolved oxygen(DO) on cold active lipase production by batch culture of metagenomic library recombinant clone(CAPL3) from deep-sea sediment were investigated. First, a two-stage temperature control strategy was developed, in which the temperature was kept at 34 °C for the first 15 h, and then switched to30 °C. The cold active lipase activity and productivity reached 315.2 U·ml^(-1)and 8.08 U·ml^(-1)·h^(-1), respectively,increased by both 14.5% compared to the results obtained with temperature controlled at 30°C. In addition, different DO control modes were conducted, based on the data obtained from the different DO control strategies and analysis of kinetics parameters at different DO levels. A step-wise temperature and DO control strategy were developed to improve lipase production, i.e., temperature and DO level were controlled at 34 °C, 30% during 0–15 h;30 °C, 30% during 15–18 h, and 30 °C, 20% during 18–39 h. With this strategy, the maximum lipase activity reached 354.6 U·ml^(-1)at 39 h, which was 28.8% higher than that achieved without temperature and DO control(275.3 U·ml^(-1)).
基金
Supported by the Hi-Tech Research and Development Program of China(863 program of China
2012AA092103)
China Ocean Mineral Resources R&D Association(DY125-15-T-06)