How do I get Sea Cucumber Protein Isolate?

"How do I get Seacucumber Protein Isolate?"

well, actually i just wanna share about my struggle on my research..
My topic research is about "the potency of sand seacucumber protein isolate in reducing blood glucose level of rats"

hmm... First of all, you have to get the seacucumbers! Thankfully, i didn't search it by myself, a Ph.D student who is also involve in this research had got seacucumbers (200 seacucumbers!) from Lampung..
Second, you have to cut them to get the flesh! Gosh, it was really hard since it need a lots, bunch bunch of energies to slice the muldy, swelly skin, throw the unnecessary parts, and cut the flesh into smaller parts.. I spent a lot of time for this!
Then, you have to blend the flesh in blender. After that, dry the flesh by freeze dryer (it was really2 expensive!)
fyi, freeze dryer is the best dryer ever! It can dry samples without destroy the structure or even reduce nutrition contained in the samples!
It needs couples days to dry the flesh.. Then, freeze-dried samples has to be blended again by blender, the use is to get smaller particles for softest flour..Then, filter the samples by 60 mesh shaking filter.. Due to lack of sample n also lack of money,I did it by myself manually *do not ever imagine it how hard this part!
Tadaaaa.... Finally, you get the seacucumber flour!! Congrats!! Since it is the raw material to be used... Lol

what else??
Hmm... Honestly, before you make protein isolate, you have to make a little research about optimum whiting protein extraction by pH adjustion.. It is because there is no literature or former research about this topic, i'm the first one! *hv no idea wanna sad or not, Hehehe
This part took 4-5 times trial till i got the best result.. Fyi, i used Lowry Method of protein. For Each, i need a whole day doing this.. From 7am till 6pm (not including washing the bottles n stuffs!!)
then, after you got which best pH you should homogenise the flour or precipitate it.. You can start making protein isolate!!

Here we go!

you have to homogenise the sea cucumber fluor by destilated water in 1: 15, then heat the solution in 40 degrees Celcius in water bath for 40 minutes. next, you should add alkaline liquid such as NaOH (sodium hydroxide) to increase its pH (till pH 11) from normal pH (6-7). the aims is to make the whole protein soluble..

After that, centrifuge the samples in 10000rpm for 15 minutes. FYI, there is no best centrifugator in my faculty so that i had to go to veterinary science faculty which is quite far from my lab to centrifuge the samples.. in addition, to bring all the samples n stuffs, you can not ride motorbike or any vehicles, afraid losing samples during the journey! it was soooooooooooo PRECIOUS!! Hmm...

Arriving in veterinary science faculty, esp in integrated science laboratory, then you can start centrifuge the samples. After first centrifugation, do not forget to divide samples into two parts, the supernatant and precipitate. take the liquid supernatant and remove the precipitate, its not necessary.. well, its can be used for feeding cat, LOL

another step!! *do not boring please~~ hehehe
add acidic liquid such as HCl to the supernatant and adjust its pH until pH 3. Based on former research I done, pH 3 is the optimum pH in precipitating sea cucumber whiting protein. Then, centrifuge in 10000rpm again for 15 minutes..
Finally, you can optimize precipitation by this adjusting!!

Take the final precipitate which is the white wetty part, put in small bottles, weight the samples and its called wet protein isolate. next (again!), you have to dry it in freeze dryer! So tough? Yes, it is!! :? *almost dying

Freeze-dried protein isolate had been ready to be used for oral ingestion to the rats! later on I will tell you how to ingest samples to the rats and how to measure blood glucose of the rats!

To be continue....