Team:SCUT-ChinaA/Description

T2


Background

Due to its unique plant aroma and potential pharmaceutical properties, the terpenoid spice has attracted attention and has a rapidly growing market demand. At present, the most important source is the direct extraction method. Due to the limitations of plant growth and other conditions, the product is difficult to purify, and the yield is low. It can not meet the industrial demand. At the same time, the chemical synthesis of terpenes is difficult to synthesize or has low yield and poor quality. The biosynthetic method of producing spices is recognized as a natural product and is currently the most important research direction for the breakthrough of the synthesis of terpenoids. However, because of various types of terpenoids, it is difficult to obtain high-yield production strains due to complex metabolic network and regulation. Therefore, the selection of a suitable host, the re-routing of the metabolic network, and the control of the direction and flow of metabolism can increase the production of terpenoids.

Overview

The literatures show that Yarrowia lipolytica is a non-conventional yeast that has the potential to become a superior terpenoid spice synthetic chassis cell. Our project aims to use the CRISPR/Cas9 technology to introduce two genes encoding neryl diphosphate synthase 1 (NDPS1) and d-limonene synthase (LS) in the synthesis of the limonene in Yarrowia lipolytica. A Catcher-Tag system is going to be applied to construct a multi-enzyme assembly system in order to reconstitute the direction and flow of the metabolites that migrate through the MVA pathway to terpenoids. Furthermore, two rate-limiting enzymes (HMG1 and ERG12) of the MVA pathway will be overexpressed to increase the MVA metabolic flux, and consequently reaches the goal of increasing production of limonene. The implementation of the project promotes the construction of a new type of limonene cell plant and provides new ideas for the study of metabolic regulation of secondary metabolites.