Phactory

CCIC

What's the E.SPA ?

Abstract

As a large agricultural country, China produces billions of tons of straw per year. The cellulose in straw is abundant and cheap, thus it’s a good carbon source. However, due to high utilization cost, low added value and low industrialization, many straws are burned, which wastes resources and also causes strong environmental pollution.

Our project hopes to construct an industrial strain that breaks down cellulose into glucose and can directly use cellulose as a carbon source. Therefore, we can provide a new idea for the utilization of straw – from field waste to industrial substrate. Additionally, considering the good production and fermentation capacity of E. coli cells, we can use this strain to produce a variety of substances. Among them, astaxanthin is the strongest natural antioxidant found in nature, which has broad market prospects and application value. By optimizing and constructing new metabolic pathways in engineered bacteria, we can achieve the conversion of glucose to astaxanthin based on cellulose as a carbon source, thus providing new ideas for the conversion of biomass to available energy.

Part

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CrtE,CrtB,CrtI and CrtY are amplified by PCR from 3 kinds of bacteria: Rhodobacter sphaeroides, Rhodospirillum rubrum and Pantoea agglomerans;

CrtZ and BKT come from Pantoea ananatis and E.coli respectively and need some modification, so we decided to synthesize this two genes chemically.Astaxanthin may be toxin to E.coli, so we choose a constitutive vector to carry our genes.

Limited by the number of suitable enzyme cleavage sites, we need to overlap CrtE,CrtB and CrtI firstly and then tap them with other genes and vector using restriction endonuclease sites.