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This is a project to make real vegan cheese by engineering normal baker's yeast (S. cerevisiae) to express milk protein (casein), purifying the protein, creating a milk-substitute by blending in vegan replacements for lactose and milkfat, and finally turning the resulting milk-substitute into semi-hard cheese like gouda using the normal cheese-making process.
The goal of this project is to make real vegan cheese by engineering normal baker's yeast (S. cerevisiae) to express milk protein (casein), purifying the protein, creating a milk-substitute by blending in vegan replacements for lactose and milkfat, and finally turning the resulting milk-substitute into semi-hard cheese like gouda using the normal cheese-making process.
* Watch a [https://www.indiegogo.com/projects/real-vegan-cheese/ brief video about the project].
* Check out our website at [https://www.realvegancheese.org/ realvegancheese.org].
* Read our [https://www.realvegancheese.org/news Blog Posts] about this project.


The main challenges involve:
The main challenges include:


* Getting the major casein-proteins secreted.
* Biology
* Achieving a high level of expression.
** Find all the nitty-gritty on how we engineer our host organism to produce cheese proteins in our pages on [[molecular biology]], [[cloning strategy]], [[gBlock design]], [[plasmid design]], [[Yeast Transformation]].
* Ensuring the required post-translational modifications take place.
** Getting the major [[Molecular_biology#Proteins|casein proteins]] expressed, and ideally secreted.
* Cost-efficient protein purification.
** Achieving a high level of expression.
* Stable micelle-formation.
** Ensuring any necessary [[Molecular_biology#Post-translational_modification|post-translational modifications]] take place.
* Finding a suitable milk-fat replacement.
** Cost-efficient [[protein purification]].
** [[Molecular_biology#Role_of_phosphorylation_and_glycosylation_in_micelle_formation_or_coagulation |Stable micelle-formation.]]
** Finding suitable [[alternative fats]].
** Finding suitable [[alternative sugars]].
** Can we achieve the same flavor and aroma profile using the traditional cheese ripening [[bacterial cultures]]?


= Project pages =
* [[Ethical, Legal, and Social Implications]]
** Can we produce this cheese in a way that is (a) better for the environment, and (b) economically feasible? Check our page on [[Life Cycle Assessments and Techno-Economic Analyses]]


*[[Vegan cheese/Molecular biology|Molecular biology]]
* Safety
*[[Vegan cheese/Cheese making|Cheese making]]
** Ensuring a safe and responsible research environment is already a key goal of CCL and BioC, but we will have to field potential safety concerns specifically related to the Real Vegan Cheese project.
*[[Vegan cheese/Ethical, Legal, and Social Implications|Ethical, Legal, and Social Implications]]
** Check out the [https://www.genspace.org/community-biology-biosafety-handbook Community Biology Safety Handbook], edited with help from some of our members.
*[[Vegan cheese/Product viability|Product viability]]


= External communications =
* Funding and spreading the word
** We have raised about $37,000 on indiegogo and the funding we raised is being used on developing the feasibility of producing Real Vegan Cheese from baker's yeast.


* Awesome foundation SF
* Patents
** Reason: $1000 grant application
** We plan to patent and abandon to keep this technology free for everyone
** Contacted by: (?)
* Awesome without borders
** Reason: $1000 grant application
** Contacted by: (?)
* [http://www.chr-hansen.com/ Chr Hansen]
** Reason: Possible sponsorship of microbial Chymosin and Lactic Acid Bacteria
** Contacted by [[User:Juul|Juul]] ([[User talk:Juul|talk]]) 17:47, 17 April 2014 (PDT)
** Result: The relevant employee is away on easter vacation. Should hear back after easter.
* [http://www.genomics.cn/ BGI]
** Access to narwhal casein protein and kinase sequences if it exists
** Contacted by [[User:Juul|Juul]] ([[User talk:Juul|talk]]) 17:47, 17 April 2014 (PDT)
** Result: They did not have narwhal. They quoted us $1500 to $2000 for full genome Narwhal mRNA sequencing.


= Shared files =
* Education
** Providing hands-on lab training.
** Check out our [[Educational Videos]] about many parts of the process!


We use [http://seafile.com/ Seafile] to share files. You can use the web interface to access our public files here:
= Getting involved =
[https://www.realvegancheese.org/news/so-you-want-to-join-the-real-vegan-cheese-project-orientation-for-new-folks '''So, you want to join the Real Vegan Cheese project? - Orientation for new folks'''] - September 2022 blog post.


*[https://files.counterculturelabs.org/d/2b8d0fe3d6/ download]
Here are some different ways to get involved:
*[https://files.counterculturelabs.org/u/d/e7fc041927/ upload]


Unfortunately many of our files are not public since most of the relevant scientific articles use closed licenses that don't allow us to share them publicly.
*Go to one of our Monday evening working meetings 7pm PST:
**[https://www.meetup.com/counter-culture-labs/events/calendar/ Counter Culture Labs Meetup calendar]
**[https://www.meetup.com/biocurious/events/calendar/ BioCurious Meetup calendar]
*Read our [[Introductory textbook|Introductory mini-textbook]] (work in progress)
*Get on one or more of our [[mailing lists]]
*[https://realvegancheese.org/contribute/ Help fund the project]
*Send us an email at info@counterculturelabs.org
*Add to this wiki.


If you want to collaborate on this project and get access to our private files, you probably want to [http://seafile.com/en/download/ install the Seafile app] (get version 2.1.x) and request an account from info@counterculturelabs.org.
= More links to incorporate =
 
*[[Science]]
'''NOTE''': When connecting with the seafile software, you need to specify https://files.counterculturelabs.org/ as the server.
**[[Microbiology]]
 
**[[Protein Expression in Yeast]]
= Mailing list =
*[[Bureaucracy]]
 
*[[Cheese making]]
The iGEM team has a mailing list but we're waiting for the admin to mark it as public so we can link it here.
*[[Existing vegan cheeses]]
 
*[[Experiments started January 2016]]
= Meeting notes =
*[[External communications]]
*[[iGEM team meeting notes 5/26/14]]
*[[Finances]] - Incoming monies, purchases, etc.
*[[iGEM team meeting notes 5/19/14]]
*[[Health effects: allergy and cancer link]]
*[[iGEM team meeting notes 5/12/14]]
*[[How to take notes and record data]]
*[[iGEM team meeting notes 5/06/14]]
*[[Media]] - Articles written about us
*[[iGEM team meeting notes 4/28/14]]
*[[Product viability]]
*[[iGEM team meeting notes 4/21/14]]
*[[Narwhal Cheese]]!
*[[iGEM team meeting notes 3/31/14]]
*[[Questions and answers]]
*[[iGEM team meeting notes 3/24/14]]
*[[Service and reagent providers]] - Sequencing, synthesis, enzymes, etc.
 
*[[Shared Laboratory Notebook]]
= Reasons why the vegan cheese project is awesome =
*[[Similar projects]] - Other people doing similar stuff
 
*[[Software]]
# It will get a lot of attention to Counter Culture Labs (great PR angle)
*[[Video]] - Video and audio documentation, teleconferencing and live-streaming
# It could be used to show the local environmentalist organizations that GMO itself is not pure evil and that we (CCL) are on their side (which I feel that we are). E.g. Sierra Club, Earth First, Greenpeace, etc.
# It can involve people who don’t have much lab experience, since we’ll have to actually make cheeses both with milk (to ensure that we can make cheese at all), with dried casein from milk and butter (to ensure that we can reconstitute the casein into something that can be used to make cheese) and with dried casein and a milkfat replacement.
# It is something that will be potentially useful over a relatively short timespan.
# It is ethically awesome
# It could turn into a commercial product
# Selling the worlds first real vegan cheese at an auction could fetch CCL quite a bit of money.
# Everyone loves cheese (ok maybe not everyone).
# Nutritional yeast is already used as a cheese substitute by many vegans, so we could make a cheaper less purified product and still have it be a viable cheese substitute.
# It is fairly realistic to at least express the protein in yeast and the project can easily be divided into separate phases with their own independent success criteria, many of which can be worked on in parallel (express protein, export protein from cell, express a lot of protein, purify protein, make cheese from protein, find viable replacement for milkfat, scale up production).
# We could label the cheese “Ethical GMO” or “Non-evil GMO” or invent our own term. Maybe a backronym Genetically Magnificent Organism. Genetically Modified and Open.
# We will have to use another sugar than lactose (probably not a problem, lactobacteria can live on other sugars), which will make our cheese lactose free.
# Marc has already built a wifi-enabled cheese cave out of a wine cooler that we can use to age our cheese prototypes.
# There is _so_ much info available on cheese chemistry because cheese industry.
# Chymosin, the rennet enzyme, is already commercially produced by genetically modified organisms. They label it “microbially produced” and most major cheeses use it so also producing the casein protein shouldn’t make any difference with regards to law/regulation.

Latest revision as of 02:48, 18 July 2023

The goal of this project is to make real vegan cheese by engineering normal baker's yeast (S. cerevisiae) to express milk protein (casein), purifying the protein, creating a milk-substitute by blending in vegan replacements for lactose and milkfat, and finally turning the resulting milk-substitute into semi-hard cheese like gouda using the normal cheese-making process.

The main challenges include:

  • Safety
    • Ensuring a safe and responsible research environment is already a key goal of CCL and BioC, but we will have to field potential safety concerns specifically related to the Real Vegan Cheese project.
    • Check out the Community Biology Safety Handbook, edited with help from some of our members.
  • Funding and spreading the word
    • We have raised about $37,000 on indiegogo and the funding we raised is being used on developing the feasibility of producing Real Vegan Cheese from baker's yeast.
  • Patents
    • We plan to patent and abandon to keep this technology free for everyone
  • Education
    • Providing hands-on lab training.
    • Check out our Educational Videos about many parts of the process!

Getting involved

So, you want to join the Real Vegan Cheese project? - Orientation for new folks - September 2022 blog post.

Here are some different ways to get involved:

More links to incorporate

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