Team:SZ SHD/Collaborations
Collaborations
NDNF China
Check out -->Here
As protein, our keratinases are known to be hard to store without a proper way. NDNF_China shares their recipe of bio-safety hydro gel with us, as we considered it a possible medium for enzyme packaging. We assayed the enzyme activity after storage and also sent a copy of the data to them.
They provided SZ-SHD with a form of the ultimate product package. SZ-SHD selected bacteria origin keratinase that degrade hair properly to make it a hair removal product. Thus we are looking for a medium able to preserve the enzymes for long-term storage and insulate them from the external environment. Through the communication with SZ-SHD, we all believe the safety, and stable condition provides for bacteria from the Hidro can also be used for keratinase. They shared the recipe of Hidro, gave us instructions for making it, and sent us some regents. Eventually, results from SZ-SHD reveals that Hidro can indeed maintain enzyme activity effectively under room temperature (25°C) storage conditions.
Hydrogel Recipe
Equipment:
5ml Syringe
Beaker
Glass rod
Autoclave sterilizer
Vacuum drier & Water Circulating Multi-purpose Vacuum Pump
Materials:
1. Viscous sodium alginate: Sigma-Aldrich A2033
2. CaCl2: Sigma-Aldrich223506
3. Acrylamide: sigma-Aldrich A8887
4. Ammonium persulfate: sigma-Aldrich A3678 4 degrees, ready to use
5. N,N-methylenebiscrylamide: sigma-Aldrich 146072 4
6. TEMED: sigma-Aldrich T9281 4
7. Nitrogen
8. 2-N-morpholino-ethane sulfonic acid (MES): Macleans
9. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) (very easily hydrolyzed) (add it at the end!) -20 seal
10. N-hydroxysuccinimide 4 degrees
11. 0.00075% adipic acid dihydrazide (ADH) -20
Solution configuration:
MES buffer: 2-N-morpholino-ethanesulfonic acid MES 9.76g and NaCl14.61g are mixed and dissolved, and the volume is adjusted to 500ml PH6.0
Cross-linking system (prepared and used now): 0.00125% 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC), 0.000375 % N-hydroxysuccinimide N-hydroxysuccinimide, and 0.00075% adipic acid dihydrazide adipic acid dihydrazide (ADH) mixed, dissolved in MES buffer
Preparation:
Alginate core
1. Alginate is dissolved in ultrapure water (deionized water) and sterilized at 120°C to obtain a 5% alginic acid solution
2. The bacterial solution is mixed 1:1 with 5% alginic acid solution, the final concentration of the solution alginic acid is 2.5%
3. Take the mixture with a syringe and drop it on the parafilm to form a bean-shaped pellet (50-100 micrometers)
4. Immerse in 5%CaCl2 for 15min to solidify
Preparation of the gel shell (4°C in the whole process)
1. Prepare a preliminary solution of 2% alginate, 30% acrylamide, 0.046% ammonium persulfate, 0.015% N,N-methylenebiscrylamide, and vacuum degassing
2. The preparation solution and tetramethylethylenediamine (TEMED) solution are mixed at 1000:1 to form a pre-gel
3. Soak the alginate core in the pre-gel to form an adjustable 100-1000μm film in a nitrogen environment. (According to the soaking time)
4. Soak the gel in step 3 in MES buffer
5. The gel in step 4 is immersed in the cross-linking system, and cross-linked in a shaker at 4°C for 3 hours
GA_State_SW_Jiaotong
Check out -->Here
For this year, team GA_State_SW_Jiaotong is aiming at bald recovery. Since we have the same target, hair, we contacted at the very beginning of the project despite the different mechanisms. We have had discussions about the safety form filling and the potential ethical problems of later experiments. Also, we propagandized each other's projects on the WeChat subscription account.
FAFU_China
Check out -->Here
FAFU developed a system of aroma production by light-regulated microorganisms, which gives microorganisms the ability to produce aroma by regulating endogenous pathways and cutting into different exogenous enzymes, and through mathematical modeling, determine its optimal culture conditions so that it stably exists in the developed system. By adding light-controlled switch elements to microorganisms, the concentration of the aroma can be changed according to the intensity of light. The inhibition of linalool synthase in E. coli is relieved under light, and linalool can be produced. Saccharomyces cerevisiae can produce nerol in dark conditions, and the related proteins are eventually degraded under the light. Finally, They designed an ingenious hardware device for the whole system, which effectively solved the problem of microbial leakage and aroma diffusion. Looking to the future, their team hopes to apply the system to the development of new nightlights and into people's lives.
We communicated and exchanged ideas since we have known their goal that automatically scents production by prokaryotic cells, while we were looking for similar substances to prevent the unpredictable odor from our catabolic enzyme. We discussed a few methods on the construction of mathematical modeling and they gave us a few suggestions on the concept of Michaelis-Menten function as well as the response surface by the provision of related essays as examples.
