Throughout our project in working with NutriPlus, each week a different department head would contact us in effort to ensure that we are on schedule and are keeping up with the workload. Following these memos ensured that our product is created with the highest quality considerations. In order to be aware of our actions, first you must be aware of what we were tasked with.

 

Memo 1: Choosing our research partners

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

The primary task for week one was to seek out a target market alongside reviewing literature in order to cement the idea. This week was also the beginning of the figurative creative juices flowing. A diverse group consisting of a diversely skilled group members was also be formed. Our choice of group members were left to chance, as only a trio of staff were available to work on this Nutriplus Nutraceuticals with the remainder of the class being part of Bioplus. Luckily, we were all diverse in our talents and experiences yet all shared the same goal, we all wanted to develop a product food derived which would benefit the health of others. Some of our initial ideas from the brainstorming session included production of cyclosporine form another fungal strain, utilizing nutrients from food waste to develop a nutraceutical vitamin drink and to use bioactive compounds from fruit & veg sources to treat disease. Unfortunately, throughout the course of the project the trio became a duo, meaning while we lost a skillset belonging to the group, new skills were thrust upon us for the remainder of the project. Originally as all three of us were in the midst of final year projects, we agreed that it was best for us all to continually communicate our thoughts and ideas through a messaging service rather than meeting weekly at a defined time as our schedules rarely intercepted.

 

When discussing our understanding of the nutraceutical industry although this isn't our fortay we collaborated the idea that nutraceuticals are functional compounds found in food and medicinal products. When reviewing literature it cannot be denied it was hard to find an article we could understand the scientific language in. We chose review a publication called "Food waste: a potential bioresource for extraction of nutraceuticals and bioactive compounds" which highlights the current issue of increased food waste resulting from industrial, agriculture and household activities with much of the emphasis being placed on fruit & vegetable waste which is most abundant in the food & agri- industry. It also highlights specifically bioactive peptides are potentially the new generation of biologically active regulators that can prevent oxidation and microbial degradation in foods and contribute to the treatment of various diseases as well. IT quite an important and interesting read as it helps raise awareness about the current problem of food waste while simultaneously highlighting the excellent potential of food waste for bioactive compound recovery for employment in developing natural therapeutic agents.

 

Memo 2: Defining our product

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Memo two tasked the group with narrowing down potential products into one on which the project would focus on, while presenting the conclusions to the Research and Development department. Initially, we just were focusing on bioactive peptides as a whole from food waste in general but as suggested to us by the R&D manager we needed to narrow down our ideas, it was too broad. From our literature review aforementioned in memo 1, bioactive peptides were highlighted as the next best thing from the bioactive compound category. While we all had greatly researched the positive effects bioactive peptides have, it was difficult to narrow down only one string of peptides that had one specific effect. They have so much beneficial properties such as antimicrobial, anticancer, antihypertensive, antidiabetic and so on meaning they could be applicable to treat a vast rang of diseases. Due to this, we took a look at the current market and the current issues in society and realized that peptides exerting anti-hypertensive effects was the product for us. However, still we needed to narrow down the source of food waste we would source these peptides from as the R&D manager said it would be best in the long run to get the peptides from a single source rather than several. We first began with fruit and veg waste however this proved not to be a viable source of bioactive peptides according to literature. Seed and beans seemed legit however the level of peptides contained in these sources still wasn't satisfactory enough for the team. Onto our next option we looked at meat waste, although initially turned off my the fact it would be meat by-products including the likes of blood, muscle tendons and so on, meat proved to ultimately be a rich source of proteins known to contain bioactive peptides. Once settled upon an idea our aim was established which was to recover antihypertensive bioactive peptides from meat waste for generation of natural ACE inhibitors fro treatment of hypertension.

 

 

Memo 3: Website development

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In our third memo received, instructions were given on how to build a website or some channel to digitally record our project and encouraging advice on the development of our ePortfolio. While it is difficult to describe what exact actions we took, I think the final product is more than enough evidence of our compliance with the memo. Francium  Gaunt,  the Chief  Manufacturing  Scientist in the Spanish plant of Nutriplus suggested several engines to document our project which included GoogleSite, WIX, Mahara, Webnode and Carbonmade. Having taking a brief visit to each, the team agreed a website  would be the most ideal approach for creating an eportfolio. I can only speak for myself, but developing a website is a lot more challenging than it seems. Thankfully Amy has a vast array of technological skills and previous experience in creating websites and so was able to create a basic structure for the not as-technologically advanced group members such as myself to navigate around. Wordpress was one website builder in consideration as well as the recommended WIX. After exploring the extent of free elements and the extent to which we could edit our website on a free contract the team settled on choosing WIX as there was alot more choice in terms of what we could add to our website to improve our e-portfolio. Initially, it was difficult to understand how to insert things into the website and it took time to get the layout together to make it look presentable as well as visually capturing and easy to navigate around. Amy looked up youtube videos and looked at reviews of WIX to aid understanding of how to use WIX properly which she then communicated this to the other group members in a meeting so they could understand how to use it aswell. It took time but after a while we each got the hang of it.

 

Aside from the website we will also document our bioprocess on USB keys, google drive as well as through images, the use of Android & Apple devices through video files.

 

 

 

Memo 4: Defining the cell line

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In our fourth memo, our chief manufacturing scientist in our Spanish plant and the head of R&D had some questions in regard to our chosen cell line. The questions presented to us and their corresponding answers are as follows:

 

 

Q1: What cell line (or equivalent) will you need to produce your product on a small scale?

 

Originally we were thinking about performing a microbial fermentation step and were thinking of  using Aspergiullus niger, a fungal cell for the production of the enzyme glucoamylase to break the cell walls of food waste which according to literature tends to give a greater peptide yield. However when we switched our fodd waste source from plant based to meat by products this was no longer applicable. Having been made aware bacterial cells are ideal due to faster replication rates & simplicity we chose an enzyme which derives from bacterial cells. We also figured directly using a protease enzyme for hydrolysis rather than performing microbial fermentation before hand for the production of the enzyme from the cell would quicken up our process and simplify things. Our chosen cell line includes two principle enzymes: SAN and Pronase. Pronase is the enzyme active in enzyme hydrolysis which originates from streptomyces griseus. After a brief discussion with the chief manufacturing scientist he made a very good point. After the protein hydrolysis the resulting DNA would cause the mixture in the vessel to become possibly to viscous for adequate mixing. With this in mind we explored DNA enzymes for breaking down resulting DNA and we came across SAN (Salt Activate Nuclease) which we chose as an additional cell line which will ensure to get rid of any unwanted DNA in order to stop the solution from getting too viscous.

 

Q2: Are there any alternatives to your selection in Q1?

 

There are many alternatives to both of these enzymes, although by clicking the button below for more detailed information will further justify our choices. Briefly, there were many different proteases we could of chosen from which are commonly utilized for protein hydrolysis. These include the likes of crude enzyme extract, alkalase, collagenase, pepsin, pronase & trypsin. We opted for Pronase because it originates from a bacterial cell strain known as streptomyces griseus, which ultimately means that it is readily available and has similar characteristics to its parent cell. Pronase is also at its optimum at a temperature of 40 degrees celsius, meaning that not too much power will be used to heat the bioreactor. It is also a non-specific enzyme meaning it would virtually break down all proteins which is beneficial for our process since meat contains an abundance of different proteins. After narrowing down possible choices, our alternative was an enzyme known as crude enzyme extract which is widely used throughout industry. Both of these enzymes were non -specific and had similar optimum temperatures and pH values, therefore the defining factor leading to our enzyme choice was the origin of the enzyme itself as crude enzyme extract originates from a mammalian source. We chose SAN as it is a nuclease enzyme activated by high salt concentrations which is compatible with our substrate since it is also relatively high in salt. Alternatives included enzymes such as DNA polymerase, ligases and DNase yet for the optimum conditions of these enzymes they required an abundant amount of chemical buffers - which we aim to stay away from. Therefore, SAN was chosen also due to its compatability with pronase as they share optimum conditions (temp: 40C, pH 8) meaning they can operate at the same time in the bioreactor.

 

Q3: Have you considered the advantages/disadvantages of the cell line you chose in Q1? What about costs?

 

The advantages and disadvantages of each enzyme were poignant in regard to our cell line choice, so in fear of repeating myself, I feel this question is already answered in Q2. Further information is available below.

 

 

 

 

 

 

 

Q4:What scale would you like to run your prototype manufacturing trial? What type of bioreactor do you need?

 

As good quality meat waste can be difficult to obtain when contacts in the meat industry are lacking, we chose to begin with a 20 litre bioreactor. It is also best to begin on a small scale when trial & erroring a process from a cost perspective. If the bioprocess and resulting products prove successful only them will we begin to upscale.

 

We will be implementing a borosilicate bioreactor with a stainless steel lid. Although stainless steel is the most abundant material used in industry for construction of bioreactors and although it confers similar advantages to glas in terms of ease of clean, chemically resistant material and reusable nature, we went with a glass bioreactor mainly due to its transparent property. Our process will be continuous and so it is prone to contamination as it is an open system. With the transparency of glass, this will allow the team to have greater control over the process as it will enable visual inspections from outside the vessel for any contaminants. Borosilicate glass was specifically chosen because unlike ordinary glass it has the capability of withstanding high temperatures upto a maximum of 515oF (melting point: 550C) and is also highly resistant to expansion and thermal shock wherefore helping compensate for the disadvantages of using glass which would be its susceptibility to breakages and thermal expansion. While it may be expensive, the advantages of using the borosilicate bioreactor heavily outweighs the disadvantages.

 

 

Q5: What ancillaries are required for your bioreactor and why?

 

The ancillaries required for our bioreactor for our bioprocess include:

• Oxygen - for assisting mixing and for our cells.

• Carbon dioxide - for gas supply into the vessel

• Glycerol - To serve as a coolant for our cooling jacket for the vessel.

• Steam - for the steam-in-place cycle for preparation of our bioreactor.

• Sterile water - for injection and for CIP cycle. 

• CIP 100 & CIP 200 - for detergent washes during our clean-in-place cycle for preparing the bioreactor.

 

While cost may be the downfall in this project, ancillaries are essential to ensure a high quality end product.

 

 

 

 

 

 

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Memo 5: Educating our staff

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

After the development of our process through the help of peer - reviewed publications, a protocol was established for our staff to follow. See below for further details of our presentation to our staff.