Abstract
Cold-active lipase production by the psychrophilic strain Rhodococcus cercidiphylli BZ22 isolated from hydrocarbon-contaminated alpine soil was investigated. Depending on the medium composition, high cell densities were observed at a temperature range of 1–10 °C in Luria–Bertani (LB) broth or 1–30 °C in Reasoner’s 2A (R2A). Maximum enzyme production was achieved at a cultivation temperature of 1–10 °C in LB medium. About 70–80 % of the secreted enzyme was bound to the cell and was highly active as a cell-immobilized lipase which exhibited good reusability; more than 60 % of the initial lipase activity was retained after five-fold reuse. The properties of the lipase produced by the investigated strain were compared with those of a mesophilic porcine pancreatic lipase (PPL). The thermal stability of the cell-immobilized bacterial lipase was higher than that of the extracellular enzyme. Highest activity was detected at 30 °C for the cell-immobilized enzyme and for PPL, while the extracellular enzyme displayed highest activity at 10–20 °C. The bacterial lipase hydrolyzed p-nitrophenyl (p-NP) esters with different acyl chain lengths (C2–C18). The highest hydrolytic activity was obtained with p-NP-butyrate (C4) as substrate, while the highest substrate affinity was obtained with p-NP-dodecanoate (C12) as substrate, indicating a clear preference of the enzyme for medium acyl chain lengths.
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Acknowledgments
The authors gratefully acknowledge financial support from the Austrian Academic Exchange Service (OEAD) in charge of the administration of the Eurasia-Pacific Uninet scholarship, National Natural Science Foundation of China (No. 31100574) and Fund from Science and Technology Department of Jilin Province (No. 20110407).
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Yu, D., Margesin, R. Partial characterization of a crude cold-active lipase from Rhodococcus cercidiphylli BZ22. Folia Microbiol 59, 439–445 (2014). https://doi.org/10.1007/s12223-014-0318-2
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DOI: https://doi.org/10.1007/s12223-014-0318-2