Researchers identify cocktail of enzymes from termites and symbionts that break down cellulose; benefits for cellulosic biofuel production
Researchers at Purdue University led by Mike Scharf, the O. Wayne Rollins/Orkin Chair in Molecular Physiology and Urban Entomology, have identified a cocktail of enzymes from the guts of termites that may be better at getting around the barriers that inhibit fuel production from woody biomass.
The findings, published in the early online version of the journal PLoS One, are the first to measure the sugar output from enzymes created by the termites themselves and the output from symbionts, small protozoa that live in termite guts and aid in digestion of woody material.
While the idea that gut symbionts are the sole contributors to termite lignocellulose digestion has remained popular and compelling, in recent years host contributions to the digestion process have become increasingly apparent. However, the degree to which host and symbiont, and host enzymes, collaborate in lignocellulose digestion remain poorly understood. Also, how digestive enzymes specifically collaborate (i.e., in additive or synergistic ways) is largely unknown. In the present study we undertook translational-genomic studies to gain unprecedented insights into digestion by the lower termite Reticulitermes flavipes and its symbiotic gut flora. We used a combination of native gut tissue preparations and recombinant enzymes derived from the host gut transcriptome to identify synergistic collaborations between host and symbiont, and also among enzymes produced exclusively by the host termite.
Our findings provide important new evidence of synergistic collaboration among enzymes in the release of fermentable monosaccharides from wood lignocellulose. These monosaccharides (glucose and pentoses) are highly relevant to second-generation bioethanol production. We also show that, although significant digestion capabilities occur in host termite tissues, catalytic tradeoffs exist that apparently favor mutualism with symbiotic lignocellulose-digesting microbes.—Scharf et al.
Scharf and his research partners separated the termite guts, testing portions that did and did not contain symbionts on sawdust to measure the sugars created.
Once the enzymes were identified, Scharf and his team worked with Chesapeake Perl, a protein production company in Maryland, to create synthetic versions. The genes responsible for creating the enzymes were inserted into a virus and fed to caterpillars, which then produce large amounts of the enzymes. Tests showed that the synthetic versions of the host termite enzymes also were very effective at releasing sugar from the biomass.
They found that the three synthetic enzymes function on different parts of the biomass. Two enzymes are responsible for the release of glucose and pentose, two different sugars. The other enzyme breaks down lignin, the rigid compound that makes up plant cell walls.
We’ve found a cocktail of enzymes that create sugars from wood. We were also able to see for the first time that the host and the symbionts can synergistically produce these sugars.—Mike Scharf
Lignin is one of the most significant barriers that blocks the access to sugars contained in biomass. Scharf said it’s possible that the enzymes derived from termites and their symbionts, as well as synthetic versions, could be more effective at removing that lignin barrier.
Despite early evidence suggesting that termite lignocellulose digestion occurs as a collaboration between host and symbionts, the idea that this process is mediated exclusively by hindgut symbionts has proliferated and remained popular. Our findings provide important new evidence showing that digestion capabilities of the host termites are significant, and also that host and symbiont synergistically contribute to release of the fermentable monosaccharide glucose from lignocellulose in the termite gut. These findings provide unique and novel glimpses into termite digestion and host-symbiont mutualism, and they are highly relevant from basic and applied perspectives. Whether host termites acquired their intrinsic capabilities of lignocellulose digestion after adapting to a eusocial lifestyle and/or successful establishment of symbiosis is a tantalizing question that will remain unresolved until further molecular evolutionary analysis of their symbiosis.
From the basic perspective of termite social evolution, by revealing catalytic and nutritional tradeoffs, these findings provide new support of previously untestable hypotheses on termite and cockroach social living and host-symbiont mutualism. From an applied perspective, by revealing synergistic enzyme combinations and synergistic outputs, our findings contribute important new information to assist in the development of novel biocatalyst technologies and strategies for producing bioethanol from 2nd generation (non-food) feedstocks. In particular, it will now be possible to test a wide range of recombinant symbiont cellulases and hemicellulases in combination with the host enzymes reported here as synergistic biocatalyst cocktails for use in industrial biomass-to-bioethanol operations.—Scharf et al.
Scharf said his laboratory and collaborators would next work on identifying the symbiont enzymes that could be combined with termite enzymes to release the greatest amount of sugars from woody material. Combining those enzymes would increase the amount of biofuel that should be available from biomass.
The US Department of Energy and Chesapeake Perl funded the research.
Scharf ME, Karl ZJ, Sethi A, Boucias DG (2011) Multiple Levels of Synergistic Collaboration in Termite Lignocellulose Digestion. PLoS ONE 6(7): e21709. doi: 10.1371/journal.pone.0021709