Expression and biological activity analysis of peptide antibiotics hPAB-β in recombinant Pichia pastoris by high-density fermentation
HOU Rui
RAO Xiancai
CONG Yanguang
CHEN Zhijin
TAN Yinling
YANG Jie
HU Fuquan
Abstract：To evaluate the expression of hPAB- β in recombinant Pichia pastori by the high density fermentation and the biological activity of hPAB-β.Engineering bacteria was fermented in fed-batch high-density fermentation in a 3.7 L fermentor at temperature 30 ℃ and pH 4.95-5.05.When the wet weight achieved 200-250 g/L, engineering bacteria was induced by methanol.After more than 50 hours induction, fermentation was terminated and the supernatant was purified by reverse phase chromatogram, gel filtration chromatogram.and high performance liquid chromatogram.The biological activity was identified by bacteriostasis assay.Engineering bacteria was induced when the wet weight achieved 235.0 g/L, 210 g/L and 264.8 g/L in 3 fermentations, and the yields of target peptide were 73.7 mg/L, 76.6 mg/L, and 74.3 mg/L in supernatant, respectively.The recombinant peptide hPAB-β purified from fermentation supernatant showed fine bacte-riostasis activity.All data imply that engineering bacteria was fermented in fed-batch high-density fermentation successfully, and which made a foundation for the large-scale production of hPAB-β.
Keywords：Peptide antibiotics hPAB-βPichia pastorisHigh-density fermentation
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 6-9,20 )