Abstract
Low-expression levels remain a challenge in the quest to use the small laccase (rSLAC) as a viable catalyst. In this study, a recombinant Pichia pastoris strain (rSLAC-GAP-AOX) producing rSLAC under both AOX and GAP promoters (located in two different plasmids) was generated and cultivated in the presence of methanol and mixed feed (methanol:glycerol). Induction with methanol resulted in a maximum laccase activity of 1200 U/L for rSLAC-GAP-AOX which was approximately 2.4-fold higher than rSLAC-AOX and 5.1-fold higher than rSLAC-GAP. The addition of methanol:glycerol in a stoichiometric ratio of 9:1 consistently improved biomass and led to a 1.5-fold increase in rSLAC production as compared to induction with methanol alone. The rSLAC removed 95% of 5 mg/L ciprofloxacin (CIP) and 99% of 100 mg/L tetracycline (TC) in the presence of a mediator. Removal of TC resulted in complete elimination of antibacterial activity while up to 48% reduction in antibacterial activity was observed when CIP was removed. Overall, the present study highlights the effectiveness of a double promoter system in enhancing SLAC production.
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The National Research Foundation (South Africa) provided financial support (Grant 105889 and 112099). Financial support was received from the Council of Scientific and Industrial Research (CSIR) for student (Deepti Yadav) scholarship.
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Yadav, D., Ranjan, B., Mchunu, N. et al. Enhancing the expression of recombinant small laccase in Pichia pastoris by a double promoter system and application in antibiotics degradation. Folia Microbiol 66, 917–930 (2021). https://doi.org/10.1007/s12223-021-00894-w
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DOI: https://doi.org/10.1007/s12223-021-00894-w