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http://dx.doi.org/10.4014/jmb.1005.05041

Optimized Medium Improves Expression and Secretion of Extremely Thermostable Bacterial Xylanase, XynB, in Kluyveromyces lactis  

Yin, Tie (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Miao, Li-Li (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Guan, Fei-Fei (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Wang, Gui-Li (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Peng, Qing (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Li, Bing-Xue (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Guan, Guo-Hua (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Li, Ying (State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University)
Publication Information
Journal of Microbiology and Biotechnology / v.20, no.11, 2010 , pp. 1471-1480 More about this Journal
Abstract
An extremely thermostable xylanase gene, xynB, from the hyperthermophilic bacterium Thermotoga maritima MSB8 was successful expressed in Kluyveromyces lactis. The response surface methodology (RSM) was also applied to optimize the medium components for the production of XynB secreted by the recombinant K. lactis. The secretion level (102 mg/l) and enzyme activity (49 U/ml) of XynB in the optimized medium (yeast extract, lactose, and urea; YLU) were much higher than those (56 mg/l, 16 U/ml) in the original medium (yeast extract, lactose, and peptone; YLP). The secretory efficiency of mature XynB was also improved when using the YLU medium. When the mRNA levels of 13 characterized secretion-related genes in the K. lactis cultured in YLP and YLU were detected using a semiquantitative RT-PCR method, the unfolded protein response (UPR)-related genes, including ero1, hac1, and kar2, were found to be up-regulated in the K. lactis cultured in YLU. Therefore, the nutrient ingredients, especially the nitrogen source, were shown to have a significant influence on the XynB secretory efficiency of the host K. lactis.
Keywords
Thermostable xylanase B; Kluyveromyces lactis; overexpression; response surface methodology; nitrogen source; secretion-related genes;
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