Will a zygomycete help solve our energy woes?

I found the headline today, “Biofuels: Fungus Use Improves Corn-to-ethanol Process” and I was curious to find out what fungus they were talking about in the article. It turns out that researchers at Iowa State University found that Rhizopus microsporus is able to grow off part of the leftovers of ethanol production called thin stillage. The reason this is so exciting is explained below:


(Rhizopus sporangium, picture taking during PMB 110L @ UC Berkeley)

The fuel is recovered by distillation, but there are about six gallons of leftovers for every gallon of fuel that’s produced. Those leftovers, known as stillage, contain solids and other organic material. Most of the solids are removed by centrifugation and dried into distillers dried grains that are sold as livestock feed, primarily for cattle.
The remaining liquid, known as thin stillage, still contains some solids, a variety of organic compounds from corn and fermentation as well as enzymes. Because the compounds and solids can interfere with ethanol production, only about 50 percent of thin stillage can be recycled back into ethanol production. The rest is evaporated and blended with distillers dried grains to produce distillers dried grains with solubles.
The researchers added a fungus, Rhizopus microsporus, to the thin stillage and found it would feed and grow. The fungus removes about 80 percent of the organic material and all of the solids in the thin stillage, allowing the water and enzymes in the thin stillage to be recycled back into production.
The fungus can also be harvested. It’s a food-grade organism that’s rich in protein, certain essential amino acids and other nutrients. It can be dried and sold as a livestock feed supplement. Or it can be blended with distillers dried grains to boost its value as a livestock feed and make it more suitable for feeding hogs and chickens.

The idea of being more efficient by saving water and producing nutritious animal feed that can produce healthier animals that produce more meat is very interesting and worthwhile. But the article never mentions that many Rhizopus species are considered pathogens and R. microsporus when paired with Burkholderia rhizoxinia, a endosymbiont that produces rhizoxin, essentially becomes the pathogen responsible for rice seedling blight. Rhizopus also can cause serious mycoses in humans (The non squeamish can search for rhizopus mycoses on google).

I am curious if this Rhizopus has any endosymbionts that could be helping it grow on stillage or what other fungi that may not be potential pathogens might be out there that could also grow on the thin stillage.

Microbial Ecology in Science

Science has a section dedicated to Microbial Ecology including a review describing microbial biogeography studying communities on the basis of trait rather than taxonomic diversity. Certainly this interlinks with metagenomic approaches well, something I’ve been thinking about more after visiting some of the folks at Montana State Thermal Biology Institute and all the increasingly massive datasets like what CAMERA provides.

Chlamy genome investigations

Chlamy coverThis month’s Genetics has a series of articles exploring the genome (published last year & freely available at Science) of the green algae Chlamydomonas reinhardtii. These manuscripts are primarily genome analyses making for a very bioinformatics focused issue of Genetics. Some of the highlights include:

Invasion of not so tasty truffles.

(Truffle picture from BBC.com)

The BBC (link) has an interesting article about a  Chinese Black truffle being found as an invasive species in Italy. The Italian’s and European truffle aficionados are worried that the Chinese Black Truffle will outcompete the Perigord Black truffle, which is supposed to be very tasty and the second most expensive truffle by weight, behind only the Piedmont White Truffle.

The scientific journal article (link) the BBC cites is present in the new phytologist and was authored by a lab from the “Dipartimento di Biologia Vegetale dell’Università di Torino. Looks like the Chinese truffle species could be a good invasive species model and also economically important.

Truffles are interesting its amazing people would pay so much for a mushroom, sadly I can’t say if one tastes better than the other since I have not had the chance to try of the truffles mentioned above.

Trichoderma reesei genome paper published

TrichodermaThe Trichoderma reesei genome paper was recently published in Nature Biotechnology from Diego Martinez at LANL with collaborators at JGI, LBNL, and others. This fungus was chosen for sequencing because it was found on canvas tents eating the cotton material suggesting it may be a good candidate for degrading cellulose plant material as part of cellulosic ethanol or other biofuels production.  The fungus also has starring roles in industrial processes like making stonewashed jeans due to its prodigious cellulase production.

The most surprising findings from the paper include the fact that there are so few members of some of the enzyme families even though this fungus is able to generate enzymes with so much cellulase activity. The authors found that there is not a significantly larger number of glucoside hydrolases which is a collection of carbohydrate degrading enzymes great for making simple sugars out of complex ones. In fact, several plant pathogens compared (Fusarium graminearum and Magnaporthe grisea) and the sake fermenting Aspergillus oryzae all have more members of this family than does.  T. reesei has almost the least (36) copies of a cellulose binding domain (CBM) of any of the filamentous ascomycete fungi.  They used the CAZyme database (carbohydrate active enzymes) database which has done a fantastic job building up profiles of different enzymes involved in carhohydrate degradation binding, and modifications.

Whether T. reesei is really the best cellulose degrading fungus is definitely an open question.  That it works well in the industrial culture that it has been utilized in is important, but there may be other species of fungi with improved cellulase activity and who may in fact have many more copies of cellulases.  So it will be good to add other fungi to the mix with quantitative information about degradation to try and glean what are the most important combination of enzymes and activities.

One technical note.  The comparison of copy number differences employed in the paper is a simple enough Chi-Squared, work that I’ve done with Matt Hahn and others include a gene family size comparison approach that also taked into account phylogenetic distances and assumes a birth-death process of gene family size change.  It would be great to apply the copy number differences through this or other approaches that just evaluate gene trees for these domains to see where the differences are significant and if they can be polarized to a particular branch of the tree.

So will this genome sequence lead to cheaper, better biofuel production? Certainly it provides an important toolkit to start systematically testing individual cellulase enzymes. It’s hard to say how fast this will make an impact, but the work of JBEI and a host of other research groups and biotech companies are going to be able to systematically test out the utility of these individual enzymes.

There is also evolutionary work by other groups on the evolution of these Hypocreales fungi trying to better define when biotrophic and heterotrophic transitions occurred to sample fungi with different lifestyles that might have different cellulase enyzmes that may not have been observed. Defining the relationships of these fungi and when and how many times transitions to lifestyles occurred to choose the most diverse fungi may be an important part of discovering novel enzymes.

Also see

Martinez, D., Berka, R.M., Henrissat, B., Saloheimo, M., Arvas, M., Baker, S.E., Chapman, J., Chertkov, O., Coutinho, P.M., Cullen, D., Danchin, E.G., Grigoriev, I.V., Harris, P., Jackson, M., Kubicek, C.P., Han, C.S., Ho, I., Larrondo, L.F., de Leon, A.L., Magnuson, J.K., Merino, S., Misra, M., Nelson, B., Putnam, N., Robbertse, B., Salamov, A.A., Schmoll, M., Terry, A., Thayer, N., Westerholm-Parvinen, A., Schoch, C.L., Yao, J., Barbote, R., Nelson, M.A., Detter, C., Bruce, D., Kuske, C.R., Xie, G., Richardson, P., Rokhsar, D.S., Lucas, S.M., Rubin, E.M., Dunn-Coleman, N., Ward, M., Brettin, T.S. (2008). Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina). Nature Biotechnology DOI: 10.1038/nbt1403

Will mushrooms save the world?

Paul Stamets thinks so and he’s done work to make this happen.  The founder of FungiPerfecti and author many books on mushroom cultivation spoke at a TED talk recently that is worth taking a look. 

We also wrote about how Paul has contributed (and donated in some cases) Pleurotus spawn as part of Dioxin cleanup in Ft Bragg, CA and cleaning up the SF Bay with hair and mushrooms

See Paul Stamets’ TED talk.

Podospora genome published

P.anserinaThe genome of Podospora anserina S mat+ strain was sequenced by Genoscope and CNRS and published recently in Genome Biology. The genome sequence data has been available for several years, but it is great to see a publication describing the findings.  The 10X genome assembly with ~10,000 genes provides an important dataset for comparisons among filamentous Sordariomycete fungi. The authors primarily focused on comparative genomics of Podospora to Neurospora crassa, the next closest model filamentous species.  Within the Sordariomycetes there are now a very interesting collection of closely related species which can be useful for applying synteny and phylogenomics approaches.

The analyses in the manuscript focused on these differences between Neurospora and Podospora identifying some key differences in carbon utilization contrasting the coprophillic (Podospora) and plant saprophyte (Neurospora).  There are several observations of gene family expansions in the Podospora genome which could be interpreted as additional enzyme capacity to break down carbon sources that are present in dung.

The genome of Neurospora has be shaped by the action of the genome defense mechanisms like RIP that has been on interpretation of the reduced number of large gene families and paucity of transposons. The authors report a surprising finding that in their analysis that despite sharing orthologs of genes that are involved in several genome defense, they in fact find fewer repetitive sequences in Podospora while it still fails to have good evidence of RIP.

Overall, these data suggest that P. anserina has experienced a fairly complex history of transposition and duplications, although it has not accumulated as many repeats as N. crassaP. anserina possesses all the orthologues of N. crassa factors necessary for gene silencing, including RIP, meiotic MSUD and also vegetative quelling, a post transcriptional gene silencing mechanism akin to RNA interference

I think this data and observations interleaves nicely with the work our group is exploring on evolution of genome of several Neurospora species which have different mating systems. The fact that the gene components that play a role in MSUD and a RIP are found in Podpospora but yet the degree of RIP and the lack of any observed meiotic silencing suggests some interesting occurrences on the Neurospora branch to be explored.  The potentially different degrees of RIP efficiency and types of mating systems (heterothallic and pseudohomothallic) among the Neurospora spp may also provide a link to understanding how RIP evolved and its role on N. crassa evolution.

Senescence in Podospora

Another aspect of Podopsora biology that isn’t touched on, is the use of the fungus as a model for senescence.  The fungus exhibits maternal senescence which involves targeted changes in the mitochondria that leads to cell death.  The evolutionary and molecular basis for this process has been of interest to many research groups and the genome sequence can provide an additional toolkit for identifying the factors involved in the apoptosis process in this filamentous fungi. Whether it will help find a real link for aging research in other eukaryotes remains to be seen, but it is a good model system for some aspects of how aging and damage to mtDNA are linked.

Espagne, E., Lespinet, O., Malagnac, F., Da Silva, C., Jaillon, O., Porcel, B.M., Couloux, A., Aury, J., et al (2008). The genome sequence of the model ascomycete fungus Podospora anserina. Genome Biology, 9(5), R77. DOI: 10.1186/gb-2008-9-5-r77

Fungal remediation of contaminated war zones

Depleted uranium (DU) from spent ammunition used in the conflicts in Iraq and the Balkans poses a health risk to the inhabitants of those regions. This paper in Current Biology from Marina Fomina et al shows that several species of fungi including one from the mycorrhizal genus Rhizopogon (a favorite subject of study for our neighbors in the Bruns lab) are capable of sequestering DU into a less mobile mineral form.

doi:10.1016/j.cub.2008.03.011

Also picked up by the BBC.

Deep EST sequencing = RNA-Seq

The transcriptional landscape of yeast has been (further) defined with Solexa sequencing in a method deemed “RNA-Seq”, but what I would call “deep EST sequencing”.  This approach for transcriptional profiling by sequencing alone is sure to be used by many labs looking for lower and more complete ways to describe and quantitate the full population of transcripts in an organism.  

Nagalakshmi, U., Wang, Z., Waern, K., Shou, C., Raha, D., Gerstein, M., Snyder, M. (2008). The Transcriptional Landscape of the Yeast Genome Defined by RNA Sequencing. Science DOI: 10.1126/science.1158441