A two-step method preparation of semaglutide through solid-phase synthesis and inclusion body expression.

Semaglutide is currently the most promising antidiabetic drug, especially for the treatment of type 2 diabetes mellitus, due to its excellent efficacy in glycemic control and weight loss. However, the production of semaglutide remains high cost, and high yield, low cost, and high purity still remains a challenge. Herein, we reported a convenient and high-yield strategy for the preparation of semaglutide through fragmented condensation coupling, involving solid-phase peptide synthesis of tetrapeptide and on-column refolding and on-column enzyme cleavage based inclusion body expression of Lys26Arg34GLP-1 (11-37) with fused protein tags in an X-Y-D4K-G pattern. The optimized N-terminal protein tag significantly boosts inclusion body expression level, while on-column refolding and on-column enzyme cleavage avoid precipitation, enhancing efficiency and yield together with one-step purification. The successful preparation of semaglutide is expected to achieve large-scale industrial production with low cost, high yield and high purity.

Design and experimental study of a polarization imaging optical system for oil spills on sea surfaces.

The marine environment is complex and changeable. To meet the urgent needs of accurate detection and identification of oil spills, a visible/infrared dual-band common-aperture polarization imaging optical system based on a defocused plane polarization detector is designed. The optical system is evaluated by ZEMAX simulation software, and we carried out the polarization imaging oil species discrimination experiment based on the split focal plane polarization detector, which proved that for some oil species that cannot be distinguished by intensity information, the polarization information can be distinguished. It verifies the feasibility of polarization detection in the discrimination of marine oil spills, which is of great practical significance in the field of marine oil spill detection.

Spraying high concentrations of chelated zinc enhances zinc biofortification in wheat grain.

BACKGROUND: Foliar application of highly concentrated ZnSO4 fertilizer improves Zn biofortification in wheat grains. However, excess ZnSO4 ·7H2 O concentration (≥5 g kg-1 , w v-1 ) has been associated with leaf burn and yield loss, necessitating Zn sources with a high threshold concentration. The aim of this study, based on a 2 year field experiment conducted on wheat cultivated in acidic and alkaline soil, was to identify a suitable Zn formulation with a high Zn concentration or efficient adjuvant to achieve optimal Zn biofortification levels without compromising agronomic performance. RESULTS: There was a continued increase in the Zn concentration in wheat grains and a decrease in grain yield with an increase in the concentration of the Zn foliar sprays in both soil types examined. Wheats treated with chelated Zn foliar sprays - Zn glycine chelate (ZnGly) and Zn-ethylenediaminetetraacetic acid (ZnEDTA) - had less foliar injury than those treated with unchelated Zn fertilizers. Furthermore, irrespective of wheat cultivars and soil types, ZnEDTA applied to wheat at a concentration of 10 g kg-1 achieved the highest grain Zn concentration without negatively affecting the wheat performance. Adjuvant type and concentration caused no significant variation in grain Zn concentration. CONCLUSION: Overall, without foliar burn, wheat treated with 10 g kg-1 ZnEDTA foliar spray had the best performance with regard to grain Zn concentration and grain yield, which could have considerable implications for Zn biofortification of wheat grain. © 2021 Society of Chemical Industry.

Various green manure-fertilizer combinations affect the soil microbial community and function in immature red soil.

Green manure application is a common practice to improve soil fertility in China. However, the impact of different green manure-fertilizer combinations on the soil microbial communities in the low-fertility immature red soil in southern China remains unclear. In this study, we conducted a pot experiment using two common green manure crops, ryegrass (Lolium perenne L.) and Chinese milk vetch (Astragalus sinicus L.), along with a fallow treatment. We also considered three combined fertilizer management strategies, including mineral, humic acid, and organic manure fertilizers. We evaluated the soil microbial biomass, activity, communities, functional prediction and their correlation with soil properties during green manure growth and incorporation periods, to assess the potential alterations caused by different green manure and fertilizer combinations. Our findings indicate that green manure application, particularly in combination with organic fertilizers, increased the alpha diversity of the soil bacterial community, while the opposite trend was observed in the fungal community. The application of green manure altered the soil microbial communities during both growth and incorporation periods, especially the taxa that participate in carbon, nitrogen and sulfur cycles. Notably, ryegrass significantly increased the relative abundance of bacterial phylum Firmicutes and fungal phylum Ascomycota, whereas Chinese milk vetch significantly stimulated the bacterial phylum Acidobacteria and fungal phylum Glomeromycota. Compared with fallow treatments, green manure application significantly increased the soil pH by 4.1%-12.4%, and microbial biomass carbon by 29.8%-72.9%, regardless of the types of combined fertilizer. Additionally, the application of green manure resulted in a 35.6%-142.6% increase in urease activity and a 65.9%-172.9% increase in ß-glucosidase activity compared to fallow treatments, while led to a 22.5%-55.6% decrease in catalase activity. Further analysis revealed that the changes in both bacterial and fungal communities positively correlated with soil pH, soil organic matter, total nitrogen and alkali hydrolyzed nitrogen contents. Moreover, the relationship between the soil microbial community and soil enzyme activities was regulated by the specific green manure species. In conclusion, our results provide insight into the effects of different green manure-fertilizer combinations on soil microorganisms and their underlying mechanisms in improving soil fertility in the low-fertility immature red soil.

Cloning, Identification, and Functional Analysis of the Chalcone Isomerase Gene from Astragalus sinicus.

Astragalus sinicus is an important winter-growing cover crop. It is widely utilized, not only as a cover crop for its benefits in fertilizing the soil but also as a landscape ground cover plant. Anthocyanins are involved in the pigmentation of plants in leaves and flowers, which is a crucial characteristic trait for A. sinicus. The formation of anthocyanins depends significantly on the enzyme chalcone isomerase (CHI). However, research on the CHI gene of A. sinicus remains unexplored. The rapid amplification of cDNA ends (RACE) approach was used in this research to clone the CHI sequence from A. sinicus (AsiCHI). The expression profiles of the AsiCHI gene in multiple tissues of A. sinicus were subsequently examined by qRT-PCR (Quantitative Real-Time PCR). Furthermore, the function of the AsiCHI was identified by the performance of ectopic expression in Arabidopsis (Arabidopsis thaliana). The outcomes revealed that the full-length cDNA of the AsiCHI gene (GeneBank: OQ870547) measured 972 bp in length and included an open reading frame of 660 bp. The encoded protein contains 219 amino acids with a molecular weight of 24.14 kDa and a theoretical isoelectric point of 5.11. In addition, the remarkable similarity between the AsiCHI protein and the CHI proteins of other Astragalus species was demonstrated by the sequence alignment and phylogenetic analysis. Moreover, the highest expression level of AsiCHI was observed in leaves and showed a positive correlation with anthocyanin content. The functional analysis further revealed that the overexpression of AsiCHI enhanced the anthocyanidin accumulation in the transgenic lines. This study provided a better understanding of AsiCHI and elucidated its role in anthocyanin production.

Chemical composition of essential oils of Litsea cubeba harvested from its distribution areas in China.

Litsea cubeba (Lour.) Pers. is a promising industrial crop with fruits rich in essential oils. The chemical composition of essential oils of L. cubeba (EOLC) were determined for fruits harvested from eight regions in China. The overall essential oil content, obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS), ranged from 3.04% to 4.56%. In total, 59 compounds were identified, the dominant components being monoterpenes (94.4-98.4%), represented mainly by neral and geranial (78.7-87.4%). D-Limonene was unexpectedly a lesser constituent (0.7-5.3%) in fruits, which differed from previous reports (6.0-14.6%). Several components were only detected in certain regions and compounds such as o-cymene and eremophilene have never before been reported in EOLC. These results demonstrate significant regional variation in the chemical composition of EOLC. This investigation provides important information with regard to the bioactivity, breeding work and industrial applications of L. cubeba.

Substrate control of sulphur utilisation and microbial stoichiometry in soil: Results of 13C, 15N, 14C, and 35S quad labelling.

Global plant sulphur (S) deficiency is increasing because of a reduction in sulphate-based fertiliser application combined with continuous S withdrawal during harvest. Here, we applied 13C, 15N, 14C, and 35S quad labelling of the S-containing amino acids cysteine (Cys) and methionine (Met) to understand S cycling and microbial S transformations in the soil. The soil microorganisms absorbed the applied Cys and Met within minutes and released SO42- within hours. The SO42- was reutilised by the MB within days. The initial microbial utilisation and SO42- release were determined by amino acid structure. Met released 2.5-fold less SO42- than Cys. The microbial biomass retained comparatively more C and S from Met than Cys. The microorganisms decomposed Cys to pyruvate and H2S whereas they converted Met to α-ketobutyrate and S-CH3. The microbial stoichiometries of C, N, and S derived from Cys and Met were balanced after 4 d by Cys-derived SO42- uptake and Met-derived CO2 release. The microbial C:N:S ratio dynamics showed rapid C utilisation and loss, stable N levels, and S accumulation. Thus, short-term organic S utilisation by soil microorganisms is determined by amino acid structure whilst long-term organic S utilisation by soil microorganisms is determined by microbially controlled stoichiometry.

[Dynamics and Runoff Losses of Nitrogen in Paddy Field Surface Water Under Combined Application of Biochar and Slow/Controlled-Release Fertilizer].

Field experiments were conducted to investigate the effects of four fertilization treatments, that is, controlled-release fertilizer (CRF), biochar combined with controlled-release fertilizer (BC+ CRF), biochar combined with stabilized fertilizer (BC+ SF), and biochar combined with controlled-release fertilizer and stabilized fertilizer (BC+ CRF+ SF), on the dynamics of the pH and nitrogen mass concentration and runoff losses of nitrogen in paddy field surface water in the Taihu Lake Basin. The results show that the average pH of the surface water decreases by 3.16%-4.48% for BC+ CRF+ SF and is in the range of 5.64-8.15. The average total nitrogen (TN) mass concentration of surface water ranges from 19.05 to 25.23 mg·L-1. A significant decrease of 4.75%-6.58% in the TN mass concentration of surface water was observed for BC+ CRF+ SF. The average ammonium (NH4+-N) and nitrate (NO3--N) mass concentration of the surface water vary from 0.01-17.26 mg·L-1 and from 0.24-3.11 mg·L-1, respectively. Inorganic nitrogen is dominated by NH4+-N in surface water. Compared with individual CRF, other treatments significantly reduce the NH4+-N mass concentration of surface water by 35.89%-48.78% and the NO3--N mass concentration of surface water by 20.54%-37.01%. The BC+ SF combination shows a significant reduction in the NH4+-N and NO3--N mass concentration of the surface water, which greatly lowers the risk of inorganic nitrogen loss via runoff. The runoff losses of TN, NH4+-N, and NO3--N are in the range of 16.24-18.09, 1.76-2.22 and 0.76-1.38 kg·hm-2, respectively. Compared with the individual CRF, the runoff loss of TN, NH4+-N, and NO3--N of other treatments is reduced. The BC+ CRF+ SF combination shows a significant reduction in the runoff losses of nitrogen, which greatly lowers the risk of nitrogen nonpoint source pollution from paddy fields.