Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, 300, Taiwan.
Microb Cell Fact. 2022 Jul 25;21(1):149. doi: 10.1186/s12934-022-01876-4.
The intracellular molecule trehalose in Saccharomyces cerevisiae may have a major protective function under extreme environmental conditions. NTH1 is one gene which expresses trehalase to degrade trehalose. Small heat shock protein 12 (HSP12 expressed) plays a role in protecting membranes and enhancing freezing stress tolerance.
An optimized S. cerevisiae CRISPR-Cpf1 genome-editing system was constructed. Multiplex genome editing using a single crRNA array was shown to be functional. NTH1 or/and HSP12 knockout in S. cerevisiae enhanced the freezing stress tolerance and improved the leavening ability after freezing and thawing.
Deleting NTH1 in the combination with deleting HSP12 would strengthen the freezing tolerance and protect the cell viability from high rates of death in longer-term freezing. It provides valuable insights for breeding novel S. cerevisiae strains for the baking industry through a more precise, speedy, and economic genome-editing system.
酿酒酵母中的细胞内分子海藻糖在极端环境条件下可能具有主要的保护功能。NTH1 是一种表达海藻糖酶以降解海藻糖的基因。小热休克蛋白 12（表达 HSP12）在保护膜和增强抗冻应激能力方面发挥作用。
构建了优化的酿酒酵母 CRISPR-Cpf1 基因组编辑系统。证明了使用单个 crRNA 阵列的多重基因组编辑是有效的。酿酒酵母中 NTH1 或/和 HSP12 的敲除增强了抗冻应激能力，并提高了冷冻和解冻后的发酵能力。
在组合中删除 NTH1 与删除 HSP12 相结合，可以增强耐冻性，并保护细胞活力，避免在长期冷冻中死亡率过高。通过更精确、更快速和更经济的基因组编辑系统，为烘焙行业培育新型酿酒酵母菌株提供了有价值的见解。
