Selected strains of Saccharomyces cerevisiae are used for commercial bioethanol production from cellulose and starch, but the high cost of exogenous enzymes for substrate hydrolysis remains a challenge. This can be addressed through consolidated bioprocessing (CBP) where S. cerevisiae strains are engineered to express recombinant glycoside hydrolases during fermentation. Looking back at numerous strategies undertaken over the past four decades to improve recombinant protein production in S. cerevisiae, it is evident that various steps in the protein production “pipeline” can be manipulated depending on the protein of interest and its anticipated application. In this review, we briefly introduce some of the strategies and highlight lessons learned with regards to improved transcription, translation, post-translational modification and protein secretion of heterologous hydrolases. We examine how host strain selection and modification, as well as enzyme compatibility, are crucial determinants for overall success. Finally, we discuss how lessons from heterologous hydrolase expression can inform modern synthetic biology and genome editing tools to provide process-ready yeast strains in future. However, it is clear that the successful expression of any particular enzyme is still unpredictable and requires a trial-and-error approach.

选定的酿酒酵母菌株用于从纤维素和淀粉进行商业生物乙醇生产，但用于底物水解的外源酶的高成本仍然是一个挑战。这可以通过整合生物加工 (CBP) 来解决，其中酿酒酵母菌株被设计为在发酵过程中表达重组糖苷水解酶。回顾过去四十年来为提高酿酒酵母重组蛋白产量而采取的众多策略很明显，蛋白质生产“管道”中的各个步骤可以根据感兴趣的蛋白质及其预期应用进行操作。在这篇综述中，我们简要介绍了一些策略，并重点介绍了在改进异源水解酶的转录、翻译、翻译后修饰和蛋白质分泌方面的经验教训。我们研究了宿主菌株的选择和修饰以及酶的兼容性如何是整体成功的关键决定因素。最后，我们讨论了异源水解酶表达的经验教训如何为现代合成生物学和基因组编辑工具提供信息，以在未来提供可加工的酵母菌株。然而，很明显，任何特定酶的成功表达仍然是不可预测的，需要反复试验的方法。