CRISPRi-Guided Metabolic Flux Engineering for Enhanced Protopanaxadiol Production in Saccharomyces cerevisiae.

Protopanaxadiol (PPD), an aglycon found in several dammarene-type ginsenosides, has high potency as a pharmaceutical. Nevertheless, application of these ginsenosides has been limited because of the high production cost due to the rare content of PPD in Panax ginseng and a long cultivation time (4-6 years). For the biological mass production of the PPD, de novo biosynthetic pathways for PPD were introduced in Saccharomyces cerevisiae and the metabolic flux toward the target molecule was restructured to avoid competition for carbon sources between native metabolic pathways and de novo biosynthetic pathways producing PPD in S. cerevisiae. Here, we report a CRISPRi (clustered regularly interspaced short palindromic repeats interference)-based customized metabolic flux system which downregulates the lanosterol (a competing metabolite of dammarenediol-II (DD-II)) synthase in S. cerevisiae. With the CRISPRi-mediated suppression of lanosterol synthase and diversion of lanosterol to DD-II and PPD in S. cerevisiae, we increased PPD production 14.4-fold in shake-flask fermentation and 5.7-fold in a long-term batch-fed fermentation.

Identifying the driving factors of water consumption from water-energy-food nexus in the Yangtze River Delta region, China.

The current water shortage in China is critical. Moreover, the water shortage has become the main bottleneck hindering sustainable economic growth. Against the background of China's dual control target of total water use and intensity, we choose the Yangtze River Delta (YRD) region as a research object, which encompasses Shanghai, Jiangsu, Zhejiang, and Anhui. Based on the perspective of water-energy-food nexus, we employ the generalized Divisia index method to decompose the change of water use into eight factors, regional economic scale effect, regional energy use scale effect, regional food production effect, regional water technology effect, regional energy technology effect, regional water-energy nexus effect, regional food-energy nexus effect, and regional water technology for food production effect, and analyze the contribution of each driver to identify the key drivers of total water use control. The results showed that the top four influencing factors are GDP, water intensity, energy consumption, and water-energy nexus in the YRD region. Regional economic scale is the first driving factor for increasing water use in the YRD region except for Zhejiang. Water intensity is the primary driving force of water-saving in Zhejiang. Energy consumption is the third driver of increasing water use in Jiangsu and Anhui. The effect of water-energy nexus is the third driving factor that affects the change of water use in Shanghai and Zhejiang. The authorities in the YRD region should vigorously develop water and energy utilization technologies to increase the water intensity and decrease energy consumption. The energy sector should decrease the water use to decrease the water-energy nexus which is also the main driving factor affecting the change of water use.