Wu Jyh-Ming, Wang Shin-Yao, Fu Wei-Chang
Department of Chemical and Materials Engineering, Chinese Culture University, 55, Hwa-Kang Rd., Yang-Ming-Shan, Taipei 111-14, Taiwan.
Int J Mol Sci. 2012 Oct 15;13(10):13212-26. doi: 10.3390/ijms131013212.
An heterologous expression of Vitreoscilla hemoglobin (VHb) for improving cell growth and recombinant protein production has been successfully demonstrated in various hosts, including Pichia pastoris. Lower temperature cultures can enhance target protein production in some studies of P. pastoris. In this study, the strategy of combining heterologous VHb expression and lower temperature cultures in P. pastoris showed that final cell density and viability of VHb+ strain at 23 °C were higher than that at 30 °C. In addition, the effects of VHb expression on recombinant β-galactosidase production and oxygen uptake rate were also higher at 23 °C than at 30 °C. Consequently, lower temperature cultures can enlarge VHb effectiveness on cell performance of P. pastoris. This is because VHb activity obtained at 23 °C cultures was twofold higher than that at 30 °C cultures, due to a different heme production. This strategy makes P. pastoris an excellent expression host particularly suitable for increasing the yields of the low-stability and aggregation-prone recombinant proteins.
在包括巴斯德毕赤酵母在内的多种宿主中，已成功证明通过异源表达透明颤菌血红蛋白（VHb）可改善细胞生长和重组蛋白生产。在一些巴斯德毕赤酵母的研究中，较低温度培养可提高目标蛋白产量。在本研究中，在巴斯德毕赤酵母中结合异源VHb表达和较低温度培养的策略表明，VHb +菌株在23℃时的最终细胞密度和活力高于30℃时。此外，VHb表达对重组β-半乳糖苷酶生产和氧摄取率的影响在23℃时也高于30℃。因此，较低温度培养可扩大VHb对巴斯德毕赤酵母细胞性能的有效性。这是因为在23℃培养时获得的VHb活性比30℃培养时高两倍，这是由于血红素产生不同。该策略使巴斯德毕赤酵母成为一种出色的表达宿主，特别适合提高低稳定性和易聚集重组蛋白的产量。
Zotero 插件