密码子优化策略在异源蛋白表达中的应用

Application of codon optimization strategy in heterologous protein expression

摘要:

酶在医疗和生物药物方面有着广泛的应用，不仅可以用来治疗各种疾病，还在临床诊断和人体健康等方面有着重要的影响。利用微生物来表达异源蛋白已经成为获取酶最简单快速的方法。为获得高浓度和高质量的异源蛋白，常用的方法是对基因序列进行密码子优化。传统的密码子优化策略主要基于密码子偏好性和GC含量，忽略了翻译动力学和代谢水平等复杂多样的变化因素。文中从基因水平、转录水平、翻译水平、翻译后水平以及代谢水平等多方面考虑出发，提供了一个较为全面的密码子优化策略，主要包括密码子偏好性、密码子协调性、密码子敏感性、调整基因序列结构以及一些其他影响因素。同时对每种策略的内容、理论支持以及应用范围等方面作了全面的总结，并将各策略的优缺点进行了系统的比较，为异源蛋白表达提供了全方位、多层次、多选择的优化策略，也为酶工业和生物药物等方面提供参考。

Abstract:

Enzymes are widely used in medical and biopharmaceuticals. They can be used not only for various disease treatments, but also clinical diagnosis. The use of microorganisms to express heterologous proteins has become the easiest and fastest way to obtain enzymes. In order to obtain high concentration and high-quality heterologous proteins, a common method is codon optimization of gene sequences. The traditional codon optimization strategy is mainly based on codon bias and GC content, ignoring complex and varied factors such as translational dynamics and metabolic levels. We provide here comprehensive codon optimization strategy based on gene level, transcriptional level, translational level, post-translational level and metabolic level, mainly including codon bias, codon harmonization, codon sensitivity, adjustment of gene sequence structure and some other influencing factors. We also summarize the aspects of strategy content, theoretical support and application. Besides, the advantages and disadvantages of each strategy are also systematically compared, providing an all-round, multi-level and multi-selection optimization strategy for heterogeneous protein expression, and also providing references for the enzyme industry and biopharmaceuticals.