|
| 酿酒酵母蔗糖转化酶Suc2分泌途径的改造 |
| Modification of the Secretion Pathway of Sucrose Invertase Suc2 in Saccharomyces cerevisiae |
| |
| DOI：doi:10.3969/j.issn.1005-7021.2026.03.002 |
| 中文关键词: 酿酒酵母（Saccharomyces cerevisiae） 蔗糖转化酶Suc2 分泌途径 二倍体 生物乙醇 菊糖 |
| 英文关键词: Saccharomyces cerevisiae sucrose invertase Suc2 secretion pathway diploid bioethanol inulin |
| 基金项目:国家自然科学基金面上项目（32072160）；辽宁省重点攻关项目（JYTZD2023029） |
|
| 摘要点击次数: 50 |
| 全文下载次数: 10 |
| 中文摘要: |
| 为提高酿酒酵母（Saccharomyces cerevisiae）在菊糖发酵中的乙醇生产效率，考察了优化分泌途径和二倍体菌株构建相结合的策略对蔗糖转化酶Suc2分泌效率和乙醇产量的影响。以BY4741和BY4742为出发菌株，利用PGK1p启动子实现囊泡运输关键蛋白Sec1和Sso2过表达，构建重组菌株BYCO与BY2CO，并通过二倍体配合获得BY3CO。将携带MFα信号肽的Suc2表达载体转化至工程菌，评估菌株SEC1、SSO2的转录水平、胞外蛋白分泌量及Suc2的分泌效率。结果显示，重组菌株SEC1和SSO2的转录水平显著上调，二倍体菌株BY3COmS的胞外蛋白分泌量较原始菌株BYmS提高216.67%，Suc2分泌效率提升86.46%。连续发酵性能分析结果表明，BY3COmS在84 h时生物量达35.00 g/L，乙醇产量100.38 g/L，较BYmS分别提高33.49%和12.61%，同时菊糖转化率提升10.32%，达到94.77%。本研究结果表明，通过强化囊泡运输通路与构建二倍体菌株的协同作用，可以显著提高Suc2的分泌效率，提升酿酒酵母的乙醇生产性能，为构建高效菊糖转化酿酒酵母工程菌提供了新的技术路径。 |
| 英文摘要: |
| To enhance the ethanol production efficiency of Saccharomyces cerevisiae in inulin fermentation, this study investigated the combined strategy of optimizing the secretion pathway and constructing diploid strains to improve the secretion efficiency of sucrose invertase（Suc2）and ethanol yield. Starting with the BY4741 and BY4742 strains, a PGK1p promoter replacement strategy was applied to construct recombinant strains BYCO and BY2CO, which overexpress the vesicle transport-related genes SEC1 and SSO2. Diploid strain BY3CO was obtained through diploid mating. Following the transformation of the engineered strains with an MFα signal peptide-tagged Suc2 expression vector, the transcription levels of SEC1 and SSO2, extracellular protein secretion, and Suc2 secretion efficiency were assessed. The results showed significant upregulation of SEC1 and SSO2 transcription in the recombinant strains. The extracellular protein secretion of the diploid strain BY3COmS increased by 216.67% compared to the original strain BYmS, and Suc2 secretion efficiency improved by 86.46%. Continuous fermentation analysis revealed that after 84 h, BY3COmS achieved a biomass of 35.00 g/L and ethanol production of 100.38 g/L, representing increases of 33.49% and 12.61%, respectively, compared to BYmS. Additionally, the inulin conversion rate improved by 10.32%, reaching 94.77%. This study demonstrates that the synergistic enhancement of vesicle transport pathways and the construction of diploid strains significantly improve Suc2 secretion efficiency and ethanol production performance in S. cerevisiae.These findings provide a novel technical approach for the development of efficient inulin-converting S. cerevisiae strains for bioethanol production. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
