Well the cat's out of the bag! Current hypothesis is azurin as the source of the #bluesoup. Will share all our results once we have more confirmation! But it's most likely not a small molecule. - ThreadSky

bielleogy.bsky.social • 101 days ago

Well the cat's out of the bag!

Current hypothesis is azurin as the source of the #bluesoup. Will share all our results once we have more confirmation! But it's most likely not a small molecule.

Reposted from Sebastian S. Cocioba

Some fun cloning of a putative azurin from our #BlueSoup today! I made 3 pairs of primers to domesticate the sequence to remove some MoClo cut sites, produce 3 fragments, and then cloned into my UPD vector to make a Golden Braid L0 part (B3-B5 position). 1/3

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chenxinli2.bsky.social•101 days ago

Curiosity question: if you add EDTA to a blue culture, would EDTA chelate away the Cu2+ ion and make the blue color go away?

bielleogy.bsky.social•101 days ago

That's one of the next-up experiments, in addition to redox efforts

chenxinli2.bsky.social•101 days ago

If it's a protein could you try Ammonium Sulfate precipitation? And look for the concentration of Ammonium Sulfate that causes the blue to precipitate?

endyaust.bsky.social•101 days ago

I see.

drmikewiser.bsky.social•101 days ago

There goes my hope for identifying smurficins.

bielleogy.bsky.social•101 days ago

Sorry to kill your dreams

drmikewiser.bsky.social•101 days ago

No, no, it's reality that's the problem, not you.

bielleogy.bsky.social•101 days ago

Yea but the messenger can be kind of a dick

drmikewiser.bsky.social•101 days ago

Eh, not from my interactions with you over the years.

bielleogy.bsky.social•101 days ago

🫶

asjerickson.com•101 days ago

Your posts are full of extremely interest things I absolutely do not understand, which makes it fascinating. Keep up the good work. I'm off to find out what blue soup is.

bielleogy.bsky.social•101 days ago

Here's a good summary:
https://www.biotechniques.com/pcr-sequencing/the-blue-soup-saga/

tallcuppajoe.bsky.social•101 days ago

Please tell me there’s a #BlueSoup session a #ASMicrobe next year

bielleogy.bsky.social•101 days ago

😆 that would have been great. "Microbes that make things blue" session.

cindylee53.bsky.social•101 days ago

I look forward to further explorations of the hypothesis! #bluesoup has been a wonderful journey in science. I hope your mother appreciates the impact and is not upset about you leaving her soup in the refrigerator so long.

brianweaver.bsky.social•101 days ago

Interesting! I feel like you'd need a ton of azurin to make the cells that blue. I'm still holding out hope for an indigo-like pigment! https://doi.org/10.1016/j.idairyj.2023.105605

bielleogy.bsky.social•101 days ago

We've mostly ruled out small molecules like indigo or indigoidine based on size and heat stability. It seems to be a chromophore bound to a protein.

chenxinli2.bsky.social•101 days ago

Could you elaborate on the molecular weight cutoff and how it was determined? Was it a desalting column and the blue was in the flow-through but not the resin?

bielleogy.bsky.social•101 days ago

Was the spin filter ones and was mostly caught between 10kDa and 30kDa

brianweaver.bsky.social•101 days ago

Got it! Thanks for cluing me in on your work so far 💙

bielleogy.bsky.social•101 days ago

Oh also because none of the blue P carnis/paracarnis producers I've seen (including those) have the gene set needed to produce either.

gamokhan.bsky.social•101 days ago

Elinne, which organism was responsible?? The algorithm in Twitter didn't help me follow the mystery and I got lost in that part

bielleogy.bsky.social•101 days ago

Pseudomonas carnis, closely related to the Blue Soymilk strain!

cwvhogue.bsky.social•101 days ago

My lab mates purified azurin from Pseudomonas aeruginosa back in the 90's. Super rich blue - you could easily see it as it moved down a chromatography column.

bielleogy.bsky.social•101 days ago

Yea the MWCO has been a pretty hopeful indicator so far.

cwvhogue.bsky.social•101 days ago

DM me an email - will send an old Ph.D. thesis with purification/apo prep/spectroscopy of 2 azurins.

cwvhogue.bsky.social•101 days ago

Also some useful notes about copper in the medium in there.

gamokhan.bsky.social•101 days ago

Thanks!!

malariameg.bsky.social•101 days ago

I am SO happy that the Blue Soup saga is continuing on Bluesky ❤️

bielleogy.bsky.social•101 days ago

It's only fitting!

I'm planning on porting over the entire thread and as many subthreads as I can when I get time.

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