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| Impact Factors on the Expression of Recombinant Human Papillomavirus 16 L1 Protein in Pichia pastoris |
| YANG Xu1,2,3,4, HUANG Wei-wei1,2,3,4, YAO Yu-feng1,2,3,4, LIU Cun-bao1,2,3,4, SUN Wen-jia1,2,3,4, BAI Hong-mei1,2,3,4, MA Yan-bing1,2,3,4 |
| Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming 650118, China |
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Abstract Optimization approaches have been shown in many published paper to be successful in improving the expression of a heterogenous gene in yeast cells, however, it is always needed to find out whether and how an approach works for the actual case. Multiple strategies were tried for the efficient expression of HPV16 L1 in Pichia pastoris, including the use of four gene versions with distinct sequence features (M16 and Y16 optimized for Pichia pastoris cells, P16 for mammalian cells, and W16 from a wild type sequence, respectively), and the employment of plasmid vectors facilitating selection of colonies with high or low gene copy numbers and host strains with different proteinase deficiency. M16 showed the highest expression, followed by Y16 and P16, whereas W16 was merely detectable. However, the codon adaptation order from high to low is sequentially Y16, M16, W16, and P16. The results indicate that the actually effective mechanism of codon optimization might not definitely attribute to codon adaptation. In addition, improving protein stability using specific proteinase-deficient host strains was also proved to be effective for L1 expression. In conclusion, the findings added useful information to our knowledge of optimizing expression of heterogeneous genes in Pichia pastoris cells.
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Received: 14 June 2017
Published: 25 October 2017
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