Acepleiadylene (APD), a nonbenzenoid nonalternant isomer of pyrene, exhibits different electronic properties from pyrene, but has been rarely studied since its first synthesis in 1956, probably due to the difficulties in synthesis and further derivatization. In this work, we revisited this long-known compound and developed a new two-step synthetic route to efficiently access APD on the gram scale. Theoretical and experimental characterizations elucidated the unique properties of APD as compared with its benzenoid isomer pyrene, particularly revealing its dipolar structure with a narrow optical gap. The functionalization of APD was demonstrated for the first time, providing doubly brominated APD as a key precursor for further π-extension. As a proof of concept, a π-extended APD and a cyclotrimer nanographene (C 48 H 24 ) were constructed, opening up new avenues to nonbenzenoid nanographenes with low HOMO−LUMO gaps.
Background
Cellulosic biomass has great potential as a renewable biofuel resource. Robust, high-performance enzymes are needed to effectively utilize this valuable resource. In this study, metatranscriptomics was used to explore the carbohydrate-active enzymes (CAZymes), especially glycoside hydrolases (GHs), present in the rumen microbiome of Hu sheep. Select CAZymes were experimentally verified and characterized after cloning and expression in
E. coli
.
Results
The metatranscriptomes of six Hu sheep rumen microbiomes yielded 42.3 Gbp of quality-checked sequence data that represented in total 2,380,783 unigenes after de novo assembling using Trinity and clustered with CD-HIT-EST. Annotation using the CAZy database revealed that 2.65% of the unigenes encoded GHs, which were assigned to 111 different CAZymes families.
Firmicutes
(18.7%) and
Bacteroidetes
(13.8%) were the major phyla to which the unigenes were taxonomically assigned. In total, 14,489 unigenes were annotated to 15 cellulase-containing GH families, with GH3, GH5 and GH9 being the predominant. From these putative cellulase-encoding unigenes, 4225 open reading frames (ORFs) were predicted to contain 2151 potential cellulase catalytic modules. Additionally, 147 ORFs were found to encode proteins that contain carbohydrate-binding modules (CBMs). Heterogeneous expression of 30 candidate cDNAs from the GH5 family in
E. coli
BL21 showed that 17 of the tested proteins had endoglucanase activity, while 7 exhibited exoglucanase activity. Interestingly, two of the GH5 proteins (Cel5A-h28 and Cel5A-h11) showed high specific activity against carboxymethylcellulose (CMC) and
p
-nitrophenyl-β-
d
-cellobioside (pNPC) (222.2 and 142.8 U/mg), respectively. The optimal pH value for activity of Cel5A-h11 and Cel5A-h28 was 6.0 for both enzymes, and optimal temperatures were 40 and 50 °C, respectively. Both enzymes retained over 70 and 60%, respectively, of their original activities after incubation at 40 °C for 60 min. However, their activities were rapidly diminished upon exposure to higher temperatures. Cel5A-h11 and Cel5A-h28 retained more than 80 and 60% of their maximal enzymatic activities after incubation for 16 h in buffered solutions in the pH range from 4.0 to 9.0.
Conclusion
The metatranscriptomic results revealed that the rumen microbiome of Hu sheep encoded a repertoire of new enzymes capable of cellulose degradation and metatranscriptomics was an effective method to discover novel cellulases for biotechnological applications.
Electronic supplementary material
The online version of this article (10.1186/s13068-019-1498-4) contains supplementary material, which is available to authorized users.
Reversible dilation of cerebral macrovascular changes could be a new feature of MELAS and a presumed reason for fluctuant CBF. It would shed new light on the mitochondrial angiopathy.
This study demonstrates the conventional and diffusional MRI features of MELAS, suggesting a model of acute stroke-like lesions in which the cortex manifest with cytotoxic edema and the subcortical area with vasogenic edema.
A healthy gut is very important for young animal development. The rumen of ruminants expands in size with the colonization of microbiota by 2 months of age. This process is promoted by alfalfa intervention. To elucidate the mechanism of this promotion, we performed transcriptomic analyses using a cohort of 23 lambs to evaluate the effects of starter diets plus alfalfa on the development of the rumen wall from the pre- to the postweaning period. The quantitative PCR analyses were used to validate selected genes that were differentially expressed in the transcriptome mapping. We found that several metabolic processes associated with rumen tissue development were affected by solid feed intake, with genes linked to the calcium signaling transduction pathway and the metabolism of pteridine-containing compounds and homocysteine metabolic process being upregulated in the group with alfalfa intervention. The results suggest that the pteridine-containing compounds and calcium signaling are targets for precise regulation of rumen development.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.