Heterologous Expression of Polygalacturonase Genes Isolated from Galactomyces citri-aurantii IJ-1 in Pichia pastoris
- Authors
- Cho, Il Jae; Yeo, In-Cheol; Lee, Nam Keun; Jung, Suk Hee; Hahm, Young Tae
- Issue Date
- Apr-2012
- Publisher
- MICROBIOLOGICAL SOCIETY KOREA
- Keywords
- polygalacturonase; Galactomyces citri-aurantii; Pichia pastoris; heterologous expression
- Citation
- JOURNAL OF MICROBIOLOGY, v.50, no.2, pp 332 - 340
- Pages
- 9
- Journal Title
- JOURNAL OF MICROBIOLOGY
- Volume
- 50
- Number
- 2
- Start Page
- 332
- End Page
- 340
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/20423
- DOI
- 10.1007/s12275-012-1290-7
- ISSN
- 1225-8873
1976-3794
- Abstract
- The objective of this work was to isolate the polygalacturonase genes of Galactomyces citri-aurantii IJ-1 harvested from rotten citrus peels and to heterologously express these genes in Pichia pastoris. Two polygalacturonase (PG) genes from G. citri-aurantii IJ-1 were obtained and tentatively named PG1 and PG2. The genes were cloned into pPICZ alpha C, and expressed in Pichia pastoris strain GS115 with a native signal peptide or the a-factor secretion signal peptide of Saccharomyces cerevisiae. All of the recombinant proteins were successfully secreted into the culture media and confirmed as a single band with a molecular weight of 35 to 38 kDa by SDS-PAGE. The specific enzyme activities of recombinant PG1 and PG2 purified by His-tag affinity resin were 4,749 and 6,719 U/mg, respectively, with an optimal pH and temperature of pH 4.0 and 50 degrees C. The Michaelis-Menten kinetic constants for PG1 and PG2, K-m, were confirmed to be 0.94 and 0.84 mM, respectively. In the presence of Mn2+, the activity of PG1 and PG2 were increased to 160.8 and 146.4% of normal levels, respectively. In contrast, Cu2+ and Fe3+ acted as strong inhibitors to the PGs.
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Collections - College of Biotechnology & Natural Resource > Department of Systems Biotechnology > 1. Journal Articles
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