Nora Luísa Czamanski, Cassiano Murilo Henrique Anzolini, Santana Ítalo Paulino, Guazzaroni María-Eugenia, Silva-Rocha Rafael, da Silva Ricardo Roberto
Cell and Molecular Biology Department, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Front Microbiol. 2023 Jan 6;13:1069443. doi: 10.3389/fmicb.2022.1069443. eCollection 2022.
The demand for robust microbial cell factories that produce valuable biomaterials while resisting stresses imposed by current bioprocesses is rapidly growing. is an emerging host that presents desirable features for bioproduction, since it can grow in a wide range of substrates and tolerate a variety of toxic compounds. To explore suitability for application as a cell factory in biorefineries, we sought to understand the transcriptional responses of this yeast when growing under experimental settings that simulated those used in biofuels-related industries. Thus, we performed RNA sequencing of the oleaginous, carotenogenic yeast in different contexts. The first ones were stress-related: two conditions of high temperature (37 and 42°C) and two ethanol concentrations (2 and 4%), while the other used the inexpensive and abundant sugarcane juice as substrate. Differential expression and functional analysis were implemented using transcriptomic data to select differentially expressed genes and enriched pathways from each set-up. A reproducible bioinformatics workflow was developed for mining new regulatory elements. We then predicted, for the first time in this yeast, binding motifs for several transcription factors, including HAC1, ARG80, RPN4, ADR1, and DAL81. Most putative transcription factors uncovered here were involved in stress responses and found in the yeast genome. Our method for motif discovery provides a new realm of possibilities in studying gene regulatory networks, not only for the emerging host , but for other organisms of biotechnological importance.
对于强大的微生物细胞工厂的需求正在迅速增长,这些工厂要能生产有价值的生物材料,同时抵抗当前生物过程所施加的压力。是一种新兴的宿主,具有生物生产所需的特性,因为它能在多种底物中生长,并耐受多种有毒化合物。为了探索其作为生物炼制中细胞工厂的适用性,我们试图了解这种酵母在模拟生物燃料相关行业所用条件下生长时的转录反应。因此,我们在不同情况下对产油、产类胡萝卜素酵母进行了RNA测序。第一种情况与压力相关:两种高温条件(37和42°C)和两种乙醇浓度(2%和4%),而另一种情况使用廉价且丰富的甘蔗汁作为底物。利用转录组数据进行差异表达和功能分析,以从每种设置中选择差异表达基因和富集途径。开发了一种可重复的生物信息学工作流程来挖掘新的调控元件。然后,我们首次在这种酵母中预测了包括HAC1、ARG80、RPN4、ADR1和DAL81在内的几种转录因子的结合基序。这里发现的大多数假定转录因子都参与应激反应,并且在酵母基因组中存在。我们的基序发现方法为研究基因调控网络提供了一个新的可能性领域,不仅适用于新兴宿主,也适用于其他具有生物技术重要性的生物体。