Studies On Trehalose Genes And Molecular Mechanism Adapted To The Extreme Environment Of The Antarctic From Pseudozyma Sp.JCC207

Trehalose is non-reducing sugar which is important for organisms, having non-specific protective effect for the living tissue and biological macromolecules.It becomes extremely high-profiled as natural substances. Trehalose can protect biofilm, membrane proteins and DNA under harsh environments such as freezing, drying, high osmotic pressure.This function makes biological have more likely to survive. That can be seen trehalose is an important functional materials of the organism.Antarctic yeast is a generic name of Pseudozyma which lives in Antarctic sea ice, sea ice edge, or other extreme environments and is the main Antarctic fungal organisms.Antarctic yeast grows and multiply at a low temperature, high salt environment for a long time, trehalose and trehalase gene played an important role in this progess.Trehalose synthase of Antarctic yeast direct the synthes and this process is an important survival mechanism for Antarctic yeast to adapt the extreme Antarctic environment. So the research of the mechanism about trehalose in Antarctic yeast at the molecular level is very necessa.ry.Pseudozyma sp. JCC207 is a typical bacterium of Antarctic yeast, its mechanism of formation of trehalose can provide a theoretical basis for the study Antarctic habitats and other species.The paper includes the following main contents:the whole genome sequencing of Pseudozyma sp JCC207; general PCR clone TPS1 of Pseudozyma sp JCC207 and analyse bioinformatics of TPS1, TPS2; Studied transcriptional regulation mechanism of TPS1 according to real-time quantitative PCR technique; Optimized the culture conditions of Pseudozyma sp. JCC207 through single factor experiment;Optimized the extract conditions of trehalose from TCA through single factor experiment, Box-Behnken design and response surface analysis; Established quadratic regression equation of factors; Explored the conservation of breeds about Pseudozyma sp. JCC207 by adding different concentrations agents, such as trehalose, glycerol, dimethyl sulfoxide;Whole genome sequencing got all genes of Pseudozyma sp JCC207, which contains the experiments required TPS1, TPS2 genes; TPS7 ORF full length is 1827 bp, encodes 608 amino acids, predicted relative molecular mass of 64.46 Da, TPS2 ORF full length is 3963 bp, encodes 1321 amino acids, predicted relative molecular mass of 144.47 KDa; qRT-PCR results showed that under high salt and low temperature TPS1 had high expression, TPS1 was sensitive for high salt and low temperature; Single factor optimized the optimum culture temperature is 25℃; initial pH in the range of 5 to 9,yeast glowed similarly;The best extraction conditions of trehalose from TCA:TCA concentration 0.6 mol/L, volume 15.16 mL, extraction time 29.76 min.Validated, trehalose extraction is consistent between the theoretical predictions and the actual value; in trehalose conservation experiments tests showed that compared with glycerol (20%) and dimethyl sulfoxide (10%) and other traditional conservation agent,0.5% trehalose solution in the short term (5 d) of the yeast has a better conservation capacity.