Consumption of methane by aerobic and anaerobic microbes governs the atmospheric level of this powerful greenhouse gas. Whereas a biochemical understanding of aerobic methanotrophy is well developed, a mechanistic understanding of anaerobic methanotrophy has been prevented by the unavailability of pure cultures. Here we report a biochemical investigation of Methanosarcina acetivorans, a methane-producing species capable of anaerobic methanotrophic growth dependent on reduction of Fe(III). Our findings support a pathway anchored by Fe(III)-dependent mechanisms for energy conservation driving endergonic reactions that are key to methanotrophic growth. The pathway is remarkably similar to pathways hypothesized for uncultured anaerobic methanotrophic archaea. The results contribute to an improved understanding of the methane cycle that is paramount to understanding human interventions influencing Earth’s climate. Finally, the pathway enables advanced development and optimization of biotechnologies converting methane to value-added products through metabolic engineering of M. acetivorans.

A large number of mammals, including humans, have intricate outer ear shapes that diffract incoming sound in a direction-and frequency-specific manner. Through this physical process, the outer ear shapes encode sound-source information into the sensory signals from each ear. Our results show that horseshoe bats could dynamically control these diffraction processes through fast nonrigid ear deformations. The bats' ear shapes can alter between extreme configurations in about 100 ms and thereby change their acoustic properties in ways that would suit different acoustic sensing tasks.

For a better understanding terpenoid volatile production in Camellia sinensis, global terpenoid synthase gene (TPS) transcription analysis was conducted based on transcriptomic data combined with terpenoid metabolic profiling under different abiotic stress conditions. Totally 80 TPS-like genes were identified. twenty-three CsTPS genes possessed a complete coding sequence and most likely were functional. The remaining 57 in the currently available database lack essential gene structure or full-length transcripts. Distinct tempo-spatial expression patterns of CsTPS genes were found in tea plants. 17 genes were substantially expressed in all the tested organs with a few exceptions. The other 17 were predominantly expressed in leaves whereas additional eight were primarily expressed in flowers. Under the treatments of cold acclimation, salt and polyethylene glycol, CsTPS67,-69 and-71 were all suppressed and the inhibited expression of many others were found in multiple stress treatments. However, methyl jasmonate resulted in the enhanced expression of the majority of CsTPS genes. These transcription data were largely validated using qPCR. Moreover, volatile terpenoid profiling with leaves, flowers and stress-treated plants revealed a general association between the abundances of mono-and sesqui-terpenoids and some CsTPS genes. These results provide vital information for future studies on CsTPS regulation of terpenoid biosynthesis. Plant terpenoids (isoprene-C5, monoterpenes-C10, sesquiterpenes-C15, diterpenes-C20, and polyterpenoids-C5xn) possess diverse functions in plant growth and development 1-7. They play significant ecological roles in the interactions between plants and stress conditions. Generally, terpenoid molecules smaller than diterpenoids are volatile and well known for their airborne signaling function, particularly against herbivore attack 8,9. High volatility of monoterpenes and sesquiterpenes enhances the flavor and aroma of crop products 10 such as tea, which is a popular beverage well known for its fragrance and aroma 11. Tea volatile terpenoids not only are defense components against insects 12 or high solar radiation 13 , but are also essential odorants of tea products with a direct influence on flavor and quality 14-16. Aroma from volatile terpenoids is one of the main sensory properties affecting tea flavor quality 17. For instance, monoterpene alcohols such as linalool and geraniol, two of the most abundant and odor active terpenoids in tea 15 , impart pleasant floral scent to green tea and black tea 17. Terpene synthases possess a characteristic catalytic function that generates multiple terpenoid products with one substrate 18 , thus collectively contributing to numerous and different structures of plant terpenoids in addition to other modifying enzymes such as uridine diphosphate (UDP)-glucosyl transferases 19,20 and P450s 21. TPSs are responsible for converting the precursors of geranyl diphosphate (GPP), isoprenyl diphosphate (IPP), farnesyl diphosphate (FPP) and geranylgeranyl diphosphat...

Reduction of the disulfide of coenzyme M and coenzyme B (CoMS–SCoB) by heterodisulfide reductases (HdrED and HdrABC) is the final step in all methanogenic pathways. Flavin-based electron bifurcation (FBEB) by soluble HdrABC homologs play additional roles in driving essential endergonic reactions at the expense of the exergonic reduction of CoMS–SCoM. In the first step of the CO2 reduction pathway, HdrABC complexed with hydrogenase or formate dehydrogenase generates reduced ferredoxin (Fdx2-) for the endergonic reduction of CO2 coupled to the exergonic reduction of CoMS–SCoB dependent on FBEB of electrons from H2 or formate. Roles for HdrABC:hydrogenase complexes are also proposed for pathways wherein the methyl group of methanol is reduced to methane with electrons from H2. The HdrABC complexes catalyze FBEB-dependent oxidation of H2 for the endergonic reduction of Fdx driven by the exergonic reduction of CoMS–SCoB. The Fdx2- supplies electrons for reduction of the methyl group to methane. In H2- independent pathways, three-fourths of the methyl groups are oxidized producing Fdx2- and reduced coenzyme F420 (F420H2). The F420H2 donates electrons for reduction of the remaining methyl groups to methane requiring transfer of electrons from Fdx2- to F420. HdrA1B1C1 is proposed to catalyze FBEB-dependent oxidation of Fdx2- for the endergonic reduction of F420 driven by the exergonic reduction of CoMS–SCoB. In H2- independent acetotrophic pathways, the methyl group of acetate is reduced to methane with electrons derived from oxidation of the carbonyl group mediated by Fdx. Electron transport involves a membrane-bound complex (Rnf) that oxidizes Fdx2- and generates a Na+ gradient driving ATP synthesis. It is postulated that F420 is reduced by Rnf requiring HdrA2B2C2 catalyzing FBEB-dependent oxidation of F420H2 for the endergonic reduction of Fdx driven by the exergonic reduction of CoMS–SCoB. The Fdx2- is recycled by Rnf and HdrA2B2C2 thereby conserving energy. The HdrA2B2C2 is also proposed to play a role in Fe(III)-dependent reverse methanogenesis. A flavin-based electron confurcating (FBEC) HdrABC complex is proposed for nitrate-dependent reverse methanogenesis in which the oxidation of CoM-SH/CoB-SH and Fdx2- is coupled to reduction of F420. The F420H2 donates electrons to a membrane complex that generates a proton gradient driving ATP synthesis.

Heterodisulfide reductases (Hdr) of the HdrABC class are ancient enzymes and a component of the anaerobic core belonging to the prokaryotic common ancestor. The ancient origin is consistent with the widespread occurrence of genes encoding putative HdrABC homologs in metabolically diverse prokaryotes predicting diverse physiological functions; however, only one HdrABC has been characterized and that was from a narrow metabolic group of obligate CO2-reducing methanogenic anaerobes (methanogens) from the domain Archaea. Here we report the biochemical characterization of an HdrABC homolog (HdrA2B2C2) from the acetate-utilizing methanogen Methanosarcina acetivorans with unusual properties structurally and functionally distinct from the only other HdrABC characterized. Homologs of the HdrA2B2C2 archetype are present in phylogenetically and metabolically diverse species from the domains Bacteria and Archaea. The expression of the individual HdrA2, HdrB2, and HdrB2C2 enzymes in Escherichia coli, and reconstitution of an active HdrA2B2C2 complex, revealed an intersubunit electron transport pathway dependent on ferredoxin or coenzyme F420 (F420H2) as an electron donor. Remarkably, HdrA2B2C2 couples the previously unknown endergonic oxidation of F420H2 and reduction of ferredoxin with the exergonic oxidation of F420H2 and reduction of the heterodisulfide of coenzyme M and coenzyme B (CoMS-SCoB). The unique electron bifurcation predicts a role for HdrA2B2C2 in Fe(III)-dependent anaerobic methane oxidation (ANME) by M. acetivorans and uncultured species from ANME environments. HdrA2B2C2, ubiquitous in acetotrophic methanogens, was shown to participate in electron transfer during acetotrophic growth of M. acetivorans and proposed to be essential for growth in the environment when acetate is limiting.