Difference between revisions of "Team:H14Z1 Hangzhou/Demonstrate"

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Revision as of 23:08, 13 October 2018

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Demonstrate

HPLC sample

Fig1.GSH standard products peak at 7.5min

Fig2.NZ9000 intracellular extracts from no-load cells

Fig3.intracellular extracts of.NZ9000/pNZ-gshF

According to the area calculation, the product was determined by internal standard method. Sample size and standard volume are mixed for testing. The results show that the sealing area is not much different at about 7.45 min, but the area of 7.75 min is half of the original. It is verified that the peak is the product peak of GSH.

Fig4.GSH concentration / cell dry weight

Comparing the GSH yields of the two groups, the first group was lower than the second group in general, and both groups had the highest GSH yields after 6 hours of induction. Up to 8.62mg/g.

HPLC detection of SAM

Fermentation conditions: when OD600=0.4 was added with different concentration of inducer, the final concentration was 20ng/ml, 50ng/ml, 100ng/ml nisin was induced and methionine at the final concentration of 1g/l was added as substrate.

a：20ng/ml b：50ng/ml c：100ng/ml

Fig5.SAM liquid phase detection chart

Fig6.SAM2 production in NZ9000/pNZ-metk cells

Fig6. shows that although there is no significant metK protein on SDS-PAGE, the intracellular SAM production is increased in varying degrees at different induction concentrations. The highest yield of 50 ng/ml is 15.2 mg/g.