An efficient peptide ligase engineered from a bamboo asparaginyl endopeptidase.

In recent years, a few asparaginyl endopeptidases (AEPs) from certain higher plants have been identified as efficient peptide ligases with wide applications in protein labeling and cyclic peptide synthesis. Recently, we developed a NanoLuc Binary Technology (NanoBiT)-based peptide ligase activity assay to identify more AEP-type peptide ligases. Herein, we screened 61 bamboo species from 16 genera using this assay and detected AEP-type peptide ligase activity in the crude extract of all tested bamboo leaves. From a popular bamboo species, Bambusa multiplex, we identified a full-length AEP-type peptide ligase candidate (BmAEP1) via transcriptomic sequencing. After its zymogen was overexpressed in Escherichia coli and self-activated in vitro, BmAEP1 displayed high peptide ligase activity, but with considerable hydrolytic activity. After site-directed mutagenesis of its ligase activity determinants, the mutant zymogen of [G238V]BmAEP1 was normally overexpressed in E. coli, but failed to activate itself. To resolve this problem, we developed a novel protease-assisted activation approach in which trypsin was used to cleave the mutant zymogen and was then conveniently removed via ion-exchange chromatography. After the noncovalently bound cap domain was dissociated from the catalytic core domain under acidic conditions, the recombinant [G238V]BmAEP1 displayed high peptide ligase activity with much lower hydrolytic activity and could efficiently catalyze inter-molecular protein ligation and intramolecular peptide cyclization. Thus, the engineered bamboo-derived peptide ligase represents a novel tool for protein labeling and cyclic peptide synthesis.

Bamboo vs. crops: an integrated emergy and economic evaluation of using bamboo to replace crops in south Sichuan Province, China.

Based on long-term monitoring conducted in Chang-ning county, a pilot site of the 'Grain for Green Program' (GFGP), an integrated emergy and economic method was applied to evaluate the dynamic ecological-economic performance of 3 kinds of bamboo systems planted on sloping farmland. The results confirmed the positive effects of all 3 kinds of bamboo systems on water conservation and soil erosion control. The benefits gained progressively increased during the first 8 years after conversion, going from 4639 to 16127 EMyuan/ha/yr on average. All three bamboo plantations were much more sustainable than common agricultural crops planted on sloping land (CP) on both the short and long-term scales with their Emergy Sustainability Index (ESI) and Emergy Index for Sustainable Development (EISD), respectively, being 14.07-325.71 and 80.35-265.80 times that of CP. However, all 3 bamboo plantations had a Net Economic Benefit (NEB) less than that of CP during the first 8 years after conversion. Even with the government-mandated ecological compensation applied, the annual NEBECs of the Bambusa rigida (BR) and Phyllostachys pubescense (PP) plantations were, respectively, 3922.03 and 7422.77 yuan/ha/yr lower than the NEB of CP. Emergy-based evaluation of ecosystem services provides an objective reference for applying ecological compensation in strategy-making, but it cannot wholly solve the economic viability problem faced by all bamboo plantations. Inter-planting annual herbs or edible fungus, such as Dictyophora echinovolvata, within bamboo forests, especially in young bamboo plantations, might be a direction for optimizing bamboo cultivation that would improve its economic viability.

Diversity and antimicrobial activity of culturable endophytic fungi isolated from moso bamboo seeds.

Bamboos, regarded as therapeutic agents in ethnomedicine, have been used to inhibit inflammation and enhance natural immunity for a long time in Asia, and there are many bamboo associated fungi with medical and edible value. In the present study, a total of 350 fungal strains were isolated from the uncommon moso bamboo (Phyllostachys edulis) seeds for the first time. The molecular diversity of these endophytic fungi was investigated and bioactive compound producers were screened for the first time. All the fungal endophytes were categorized into 69 morphotypes according to culturable characteristics and their internal transcriber spacer (ITS) regions were analyzed by BLAST search with the NCBI database. The fungal isolates showed high diversity and were divided in Ascomycota (98.0%) and Basidiomycota (2.0%), including at least 19 genera in nine orders. Four particular genera were considered to be newly recorded bambusicolous fungi, including Leptosphaerulina, Simplicillium, Sebacina and an unknown genus in Basidiomycetes. Furthermore, inhibitory effects against clinical pathogens and phytopathogens were screened preliminarily and strains B09 (Cladosporium sp.), B34 (Curvularia sp.), B35 (undefined genus 1), B38 (Penicillium sp.) and zzz816 (Shiraia sp.) displayed broad-spectrum activity against clinical bacteria and yeasts by the agar diffusion method. The crude extracts of isolates B09, B34, B35, B38 and zzz816 under submerged fermentation, also demonstrated various levels of bioactivities against bambusicolous pathogenic fungi. This study is the first report on the antimicrobial activity of endophytic fungi associated with moso bamboo seeds, and the results show that they could be exploited as a potential source of bioactive compounds and plant defense activators. In addition, it is the first time that strains of Shiraia sp. have been isolated and cultured from moso bamboo seeds, and one of them (zzz816) could produce hypocrellin A at high yield, which is significantly different from the other strains published.

[Soil heavy metals concentration in different type plantations in Harbin City].

In order to understand the soil heavy metals characteristics in different type plantations in Harbin City, 0-10 cm and 10-30 cm soil samples were collected from eight mono-cultured plantations (Larix gmelinii, Pinus sylvestris var. mongolica, Pinus tabulaeformis var. mukdensis, Phellodendron amurense, Juglans mandshurica, Fraxinus mandshurica, Betula platyphylla, and Quercus mongolica) and one mixed plantation (P. sylvestris var. mongolica + F. mandshurica + Pinus koraiensis + P. amurense + B. platyphylla) established in the Urban Forestry Demonstration Research Base of Northeast Forestry University in the 1950s to analyze the concentrations of Cu, Zn, Cd, Ni, Pb, and As, and the comprehensive pollution index was used to assess the soil heavy metals pollution in the plantations, based on the background values of black soil in the City. There existed significant differences in the heavy metals concentration in the same soil layers among different type plantations. In the test plantations, the concentrations of soil heavy metals except As and Ni tended to decrease with soil depth. In the same soil layers, Zn had the highest concentration (62.29-126.35 mg x kg(-1)), while Cd had the lowest one (0.06-0.47 mg x kg(-1)). The abundance and accumulation tendency of test heavy metals in the plantations was in the order of Zn>Pb>Cu(Ni)> Ni(Cu)>As>Cd, and Cd>Pb>Zn>Cu>Ni>As, respectively. There existed positive correlations among soil Cu, Zn, Pb, and Cd concentrations in the plantations except F. mandshurica, and the concentrations of these four heavy metals in the plantations except F. mandshurica and Q. mongolica were positively correlated with soil organic matter, N, P, and K. The concentrations of soil Cd, Pb, Zn, and Cu were higher than the background values, suggesting the anthropogenic inputs; while those of Ni and As were similar to or lower than the background values, suggesting the origin from soil materials. The soil comprehensive pollution index of the plantations was decreased in the order of F. mandshurica > P. amurense > mixed plantation > J. mandshurica > P. sylvestris var. mongolica > P. tabulaeformis var. mukdensis > L. gmelinii >B. platyphylla > Q. mongolica.

[Effects of different type urban forest plantations on soil fertility].

Aimed to study the effects of different urban forest plantations on soil fertility, soil samples were collected from eight mono-cultured plantations (Larix gmelinii, Pinus sylvestris var. mongolica, Pinus tabulaeformis var. mukdensis, Phellodendron amurense, Juglans mandshurica, Fraxinus mandshurica, Betula platyphylla, and Quercus mongolica) and one mixed plantation (P. sylvestris var. mongolica + F. mandshurica + Picea koraiensis + P. amurense + B. platyphylla) established in Northeast Forestry University's Urban Forestry Demonstration Research Base in the 1950s, with two sites of neighboring farmland and abandoned farmland as the control. The soils in broadleaved forest plantations except Q. mongolica were near neutral, those in mixed plantation, L. gmelinii, P. sylvestris var. mongolica, and P. tabulaeformis var. mukdensis were slightly acidic, and that in Q. mongolica was acidic. The contents of soil organic matter, total N and P, available P and K, and hydrolysable N tended to decrease with soil depth. There existed significant differences in the chemical indices of the same soil layers among different plantations. The soil fertility was decreased in the order of F. mandshurica > P. amurense > mixed plantation > J. mandshurica > B. platyphylla > abandoned farmland > farmland > P. sylvestris var. mongolica > L. gmelinii > Q. mongolica > P. tabulaeformis var. mukdensis, suggesting that the soil fertility in broadleaved forest plantations except Q. mongolica and in mixed plantation increased, while that in needle-leaved forest plantations tended to decrease.