Improved natural food colorant production in the filamentous fungus Monascus ruber using CRISPR-based engineering

Hye Ree Yoon; Suk Han; Seung Chul Shin; Su Cheong Yeom; Hyo Jin Kim

doi:10.1016/j.foodres.2023.112651

Improved natural food colorant production in the filamentous fungus Monascus ruber using CRISPR-based engineering

Food Res Int. 2023 May:167:112651. doi: 10.1016/j.foodres.2023.112651. Epub 2023 Mar 1.

Authors

Hye Ree Yoon¹, Suk Han², Seung Chul Shin³, Su Cheong Yeom⁴, Hyo Jin Kim⁵

Affiliations

¹ Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang 25354, Republic of Korea. Electronic address: hyo225@snu.ac.kr.
² Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang 25354, Republic of Korea. Electronic address: fami1y@snu.ac.kr.
³ Division of Polar Life Sciences, Korea Polar Research Institute, Incheon 21990, Republic of Korea. Electronic address: ssc@kopri.re.kr.
⁴ Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang 25354, Republic of Korea; Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Republic of Korea. Electronic address: scyeom@snu.ac.kr.
⁵ Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang 25354, Republic of Korea; Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Republic of Korea. Electronic address: erythritol@snu.ac.kr.

PMID: 37087240
DOI: 10.1016/j.foodres.2023.112651

Abstract

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.

Keywords: CRISPR/Cas9; Fungal metabolic engineering; Genome editing; Monascus pigment; Monascus ruber; Natural colorant production.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Food Coloring Agents* / metabolism
Monascus* / genetics
Monascus* / metabolism

Substances

Food Coloring Agents

Supplementary concepts

Monascus ruber