Science Buddies: "Ask an Expert"

Dear Expert: I am a high school student and am interested in the Science Buddies project: Turn Plants into Biofuel with the Power of Enzymes by Sandra Slutz, PhD. I have researched the project and read numerous on-line resources, I have all the necessary equipment for fermentation and distillation and I also have access to a university chemistry lab. My starting point for my Science Fair Experiment would ideally be at the very beginning: breaking down plant fiber- waste wood products/woody material such as sawdust/wood chips. I am hoping that the Science Buddies “Power of Enzymes” project can help me complete my Science Fair Experiment
The “Power of Enzymes” project requires the Bio-Rad Biofuel Enzyme Kit, and I have read all instructions & manuals in the enzyme kit (Instruction Manual, Biofuel Enzyme Kit, Biotechnology Explorer, Rev B-A)
The kit says: that there is 3 steps in the production of ethanol, and my main problem is in this 1st step or pretreatment phase: “The Removal of non-cellulose biomolecules such as lignin”. The kit talks about step 1. The pretreatment phase and how the cellulose must be isolated from plant materials (lignins) by using heat treatment, physical crushing, acid or base treatment… but nowhere in the Power of Enzymes Kit do I see instructions using heat or acid/base treatments, to remove the lignins, which is necessary before the enzymatic hydrolysis. Am I misinterpreting the information?
Dear Expert, in your knowledgeable opinion, “can a student start this project at the very beginning? Does the kit remove the lignin from waste wood materials? The kit mentions that …once the lignin is removed, the cellulose is more exposed and can be broken down into glucose, in three steps, by three different types of enzymes. This kit provides an in-depth instruction about cellobiase, one of the enzymes used for the production of ethanol from cellulose, but I need to understand the pretreatment phase and the removal of lignin.
I have almost 7 months to complete my Science Fair Experiment/ project and ideally I would like to start with waste wood (paper, cardboard, wood, and other fibrous plant material) and I need to remove the lignin. Can you advise me how to do this? Thank you Joanne & Rebecca

Hi,
The kit you have does not remove lignin from waste wood materials. The kit is designed as an experiment to explore the effect of cellobiase on step 2, so it is starting past the step 1 pretreatment step.

If you wanted to do an experiment that starts from the beginning, you would need to devise a pre-treatment process. I think this could be done and you might want to do some research on methods of pre-treatment that could be used for the experiment. Some examples might be to cut the material up using scissors, or grind it up using a coffee grinder, and/or boil it in water.
Below is a link to a lab epxeriment that sounds similar to what you are interested in, and might be helpful:
https://www.glbrc.org/education/classro ... ss-ethanol

Good luck with your project, and let us know if you have more questions.

Thank you so much for your valuable information. The link you provided was very helpful. Thanks to you and Science Buddies, I'm off to a great start!

Hello again. In the fall I requested info on how to get me started with converting cellulose biomass to cellulosic ethanol. I was very successful with my project... Although I was not able to obtain the cellulase enzyme since I'm a private individual (not a big energy company) and also because I live in Canada (here it's not available). I instead extracted my own cellobiase proteins (enzymes) from King Oyster and Shiitake Mushrooms. I did this at my local university who allowed me access to the biochem lab even though I'm only in grade 11. After isolating the enzyme I used it to degrade and hydrolyze several types of cellulose biomass (sawdust, cardboard, sunflower stalk) that I pretreated by grinding (mechanical) as well as heat pretreatment. I did this in one test tube. One test tube for each biomas, and did a one-step simultaneous saccrinifacation and fermentation process. My results were excellent, and I ran numerous trials for each biomass. Cardboard produced the best result at 165% ethanol. My whole project started simply because I saw the "Flex-Fuel" green leaf auto symbol on the trunk of my mom's car.... but from that I went to recently winning my regional science fair here in Nova Scotia. I am now preparing for the National Canada Wide Science Fair (CWSF) in Montreal, Quebec, Canada. I've been asked the question..."what does the Industrial Cellulase sell for?" I honestly can not find the answer to that question even though I've written upwards of 10 letters to various corporations. Can Science Buddies help me please?

Hi JoanneFord,

Wow! It sounds like you've done some fantastic work here. Congratulations on your successes.

I did some checking with companies that I've purchased chemicals from in the past. You can get a laboratory grade bottle of cellulase from Carolina Biologicals for $34 (US) for a 25 gram bottle. Here's a link:

http://www.carolina.com/catalog/detail. ... AkRC8P8HAQ

The price might be lower if a company were purchasing tonnes of cellulase at a time. However, $1.36 US/gram is a value you could reference, based on that bottle that Carolina sells.

Post back if you have other questions! And best of luck in your coming competitions!