Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang 25354, Republic of Korea.
Division of Polar Life Sciences, Korea Polar Research Institute, Incheon 21990, Republic of Korea.
Food Res Int. 2023 May;167:112651. doi: 10.1016/j.foodres.2023.112651. Epub 2023 Mar 1.
Monascus pigments have various food industry applications and are pharmacologically active. Genome sequencing-based clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology has been implemented to increase pigment production in Monascus. To increase pigment production in M. ruber KACC46666, the CRISPR/Cas9 system was used to introduce mutations in two negative regulator genes (MpigI and MpigI'), among other genes involved in the Monascus pigment biosynthesis pathway. Dual single-guide RNAs were constructed to inactivate MpigI and MpigI'. After CRISPR/Cas9 inactivation, yellow, orange, and red pigment expression in the resulting △MpigI16-7 strain (among several Cas9-mediated mutants studied) was 2.5-, 12.4-, and 18.5-fold, respectively, higher than that in the wild-type strain. This study provides valuable information regarding CRISPR-guided metabolic engineering for natural colorant production.
红曲菌色素在食品工业中有多种应用，且具有药理活性。基于基因组测序的成簇规律间隔短回文重复序列（CRISPR）/CRISPR 相关蛋白 9（Cas9）技术已被用于提高红曲菌的色素产量。为了提高红曲菌 KACC46666 的色素产量，CRISPR/Cas9 系统被用于在参与红曲菌色素生物合成途径的其他基因中引入两个负调控基因（MpigI 和 MpigI'）的突变。构建了两个双单链向导 RNA 以失活 MpigI 和 MpigI'。在 CRISPR/Cas9 失活后，在产生的 △MpigI16-7 菌株（在所研究的几种 Cas9 介导的突变体中）中，黄色、橙色和红色色素的表达分别比野生型菌株高 2.5 倍、12.4 倍和 18.5 倍。这项研究为天然色素生产的 CRISPR 指导代谢工程提供了有价值的信息。
