Today Ryan and I had the privilege of shadowing each other. He did not take Bio 115, but he did catch on quickly when I showed him and explained to him what I was doing. I went into his lab absolutely clueless. I knew he was doing something with biodiesel, but was not sure what he was actually doing. He is currently comparing yields of biodiesel based on what method he used to see which one is most efficient. I observed him immobilize an enzyme onto celite. He told me that this not only increases enzymatic activity, but it is also reusable during the activity. The enzyme attaches to the celite and is filtered out through vacuum filtration leaving it for Ryan to reuse again.
Ryan previously made a sodium phosphate buffer to help dissolve (He uses 1 L for this process).
Using liquid nitrogen, Ryan performs lyophilizing (a process known as flash freezing). It is mainly a dehydration process that Ryan uses in order to allow the frozen water to sublime and go from a solid to a gas. While I was there he was in the process of freezing, but I did not stay long enough to see him add heat to it, thus converting his product.
Ryan also uses a Roto-back vacuum for his enzyme. This works as a sublimation in which all water is sucked out leaving his enzyme as a dry powder (Celite, enzyme, and salt remain). The enzyme at this stage is attached to the celite.
Ryan then uses the immobilized enzyme and incubates it with 95% ethanol and soybean oil. He sets it out on a watchglass to evaporate which he said is mainly a timing issue. Then he boils out all the ethanol (alcohol boils at a lower temperature compared to oils). He spoke of the importance of using boileezers (that should bring back some organic chemistry memories). Once this has been done, Ryan can then perform vacuum filtration in which oils are sucked in. Then he boils the enzyme and watches for the temperature to change. Once the temperature rises above 100 degree Celsius, he knows it just contains oil. Ryan also explained how weird it was that if you boil it, it turns brown…but if you evaporate it, it stays a clear color. This is just one unique physical property.
After spending some time in the inorganic lab today I am not so sure I would enjoy it. I could understand what Ryan was doing, but I was very uninterested in it mainly because I was not real familiar with his study. He was patient with me as I asked him many questions and he explained every technique thoroughly which was helpful in fully grasping what he was doing. I kind of realized how “timing is everything” for his research as well. I now have a fuller understanding for what he is doing and how beneficial this type of research is to the science community.
Cara,
ReplyDeleteIn what ways do you feel like Ryan approaches his research in the same ways that you do, and what differences do you see? I'd like to read a little more reflection on other similarities and differences of your projects based on your observations and discussions.
-kc
A lot of his research is based on time as well as mine. He works on one for 10-15 minutes and then while it is in the Roto-back he performs another activity...whereas mine might be waiting for new virgin flies after they have been cleared. He uses a lot of equipment, most of which I am unfmailiar with, in his testing whereas mine is not as equipment based. We both focus a lot on measurements which is extremely important in order to keep everything standard and consistent.
ReplyDeleteCara,
ReplyDeleteI guess Ryan had a hot topic to shadow! I too shadowed him when he did the techinique to described above. What did you think of the filtration system? To me it looked like something you would see in a museum to me. I was also interested in the fact that his enzyme comes in a powdered form, I had the idea going into the shadowing that it would be in a liquid form. Do you think you could become more interested in his doing the same kind of research if you knew more about the research background?
Casey
Casey,
ReplyDeleteHmmm? A museum? I was not thinking that...but sure! The filtration system was unique, but I feel that I would not be patient enough with it. I was intrigued by the powder form of the enzyme too and perhaps Ryan will share more info. on that in his presentation. I do think that if I knew more about his research I would enjoy it...just not the waiting part!
Cara,
ReplyDeleteDid you get chance to ask Ryan why he actually used 95% alcohol and soyabean oil when he incubated his immobilized enzyme? I am more anxious to know what a vegetable oil is doing here rather than an alcohol
Rupesh,
ReplyDeleteWell to answer that question for Cara because we never really talked about that during the incubation phase is when the triglycerides from the vegetable oil are going through transesterification and trading the methyl ester chains(biodiesel)that are on the triglycerides for the alcohol and that is how the biodiesel is formed, the enzyme that is present during this is just being used as the catalyst for this event to happen
Ryan,
ReplyDeleteI never asked you this...but are you doing your experiment multiple times with each different method and vegetable oil to make sure that the trials are actually creditable?