Team:XHD-Wuhan-Pro-China/FutureWork
1. The comparisons between our products and other alcoholics.
Other:
1) Natural Chinese herbal medicine:
particle that has the effect of curing alcohol, oral liquid that can cure alcohol and protect liver and so on are developed with some Chinese herbal medicines with the effect of relieving liquor (including clearing heat and detoxifying, nourishing liver and protecting kidney)
2) Chemical synthesis :
(1) some chemicals can enhance the metabolism of ethanol by increasing the gene expression and enzyme activity of ADH and ALDH.
(2) There are also drugs used to treat alcohol use disorders (disulfiram, naltrexone, Acamprosate), but these drugs can easily lead to drug addiction.
3) Proteins :
(1) these products use protein to form a protective layer on the stomach wall to prevent the stomach from absorbing excessive alcohol, and do not really achieve the effect of hangover relief.
(2) Direct exogenous injection of ADH and ALDH (or injection of ADH and ALDH wrapped in red blood cells) to promote alcohol metabolism by providing ADH and ALDH. However, the disadvantage of this approach is that purified ADH and ALDH are expensive and prone to induce an immune response.
4) Peptide:
It is specially designed to degrade alcohol by using small molecule peptide with its cure alcohol and protect the liver effects, which can reduce the accumulation of alcohol in the body.
We:
In addition to genes expressing ethanol and acetaldehyde dehydrogenase, our products also add NAD synthase (nadE) and NADH oxidase genes to promote the synthesis of NAD+, improve the imbalance of NAD+/NADH ratio caused by ethanol and acetaldehyde metabolism, and accelerate the metabolism of ethanol and acetaldehyde. It can also replenish the NAD+ needed for the Krebs cycle to avoid insufficient supply of cell capacity.
2. Enzyme activity of ethanol and aldehyde dehydrogenase and content of coenzyme NAD+
Firstly, crude enzyme extracts of wild type E. coli 1917 and two engineering strains were prepared respectively. Several kinds of bacteria were inoculated into LB medium with 1% inoculation amount, and cultured at 200 RPM at 37℃ until OD600 reached 1. At this time, the number of bacterial colonies in the medium was about 5 × 108-109 cfu/mL. At this time, 10 mL bacterial solution was taken and centrifuged at 5000g at 4℃ for 10 min, and the supernatant was discarded. The supernatant was discarded, and this step was repeated three times. A Binding Buffer with a PH of 7.4 (containing 1 M Tris, 5 M NaCl, and 2 M imidazole per liter) was used to suspend 10 mL of the cells. The cells were broken by an ultrasonic cell crushing apparatus, and centrifuged at 5000g at 4℃ for 15 min. The supernatant was extracted to remove insoluble cell fragments.
Ethanol dehydrogenase (ADH) test kit, acetaldehyde dehydrogenase (ALDH) activity test box, coenzyme ⅰ NAD (H) content test box and NADH oxidase (nox) test box were used to detect the activity of ethanol dehydrogenase, acetaldehyde dehydrogenase and NADH oxidase (nox) in the prepared crude enzyme extracts. And coenzyme iNAD (H).
3. In the field of microbiology, the average biotech company needs to raise large sums of money to fund a series of clinical trials before the US Food and Drug Administration can approve it.
Ethanol dehydrogenase (ADH) test kit, acetaldehyde dehydrogenase (ALDH) activity test box, coenzyme ⅰ NAD (H) content test box and NADH oxidase (nox) test box were used to detect the activity of ethanol dehydrogenase, acetaldehyde dehydrogenase and NADH oxidase (nox) in the prepared crude enzyme extracts. And coenzyme iNAD (H).
However, if we turn the other direction and let our probiotics be considered food, not medicine, we do not need to go through many clinical trials to prove that our products work in humans. At the same time, when promoting our products, we need to avoid using topics like "curing disease. “This can effectively avoid being treated as a drug by the Drug Administration. Our products can be marketed directly to consumers.