Batch and Continuous-Flow Electrochemical Geminal Difluorination of Indeno[1,2-c]furans.

An electrochemical gem-difluorination of indeno[1,2-c]furans using commercially available and easy-to-use triethylamine trihydrofluoride as both the electrolyte and fluorinating agent was developed. Remarkably, different reaction pathways of indeno[1,2-c]furans, i.e., paired electrolysis and net oxidation, are operative in a batch reactor and a continuous-flow microreactor to afford the corresponding gem-difluorinated indanones and indenones, respectively.

Emerging organophosphite and organophosphate esters in takeaway food and the implications for human exposure.

Takeaway food has become a prominent component of the diet in urban areas of China, especially for young people. Although dietary intake is a major pathway to contaminants for human exposure, studies on emerging organophosphite antioxidants (OPAs) and organophosphate esters (OPEs) in food are scarce. Here, we investigated four OPAs and 19 OPEs in takeaway foods (n = 99) and paired takeaway food packaging (n = 50) in China. AO168=O (mean: 14.9 ng/g ww), TPPO (mean: 1.05 ng/g ww), and TCIPP (mean: 0.579 ng/g ww) were dominant in the takeaway food. Some OPEs had significant correlations in takeaway food. Emerging OPAs and OPEs in takeaway food varied significantly depending on the packaging materials and food types. AO168 and AO168=O were widespread in the paired takeaway food packaging. The migration efficiencies of emerging OPAs and OPEs were low in takeaway food packaged in aluminum foil. Although the actual contamination of emerging OPAs and OPEs in takeaway food significantly differed from those of in food simulants migrated from paired takeaway food packaging, the results imply that food itself and takeaway food packaging are potential contamination sources of emerging OPAs and OPEs in takeaway food. The average estimated dietary intakes of emerging OPAs and OPEs were 465 ng/kg body weight (bw)/day and 91.9 ng/kg bw/day, respectively. The exposure risk of emerging OPAs and OPEs through takeaway food intake is low in China.

Solibacillus ferritrahens sp. nov., a novel siderophore-producing bacterium isolated from Wumeng Mountain National Nature Reserve in Yunnan Province.

A gram-stain-positive, aerobic, rod-shaped bacterial strain capable of producing siderophores, named YIM B08730T, was isolated from a soil sample collected from Wumeng Mountain National Nature Reserve, Zhaotong City, Yunnan Province. Growth occurred at 10-45 °C (optimum, 35-40 â„ƒ), pH 7.0-9.0 (optimum, 7.0) and in the presence of 0-5 % (w/v) NaCl (optimum, 0-1 %, w/v). A comparative analysis of the 16S rRNA gene sequence (1558 bp) of strain YIM B08730T showed the highest similarity to Solibacillus isronensis JCM 13838T (96.2 %), followed by Solibacillus silvestris DSM 12223T (96.0 %) and Solibacillus kalamii ISSFR-015T (95.4 %). The main polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and one unidentified lipid. The main respiratory quinone of strain YIM B08730T was menaquinone 7 (MK-7). The major fatty acids were iso-C15:0 and C16:1ω7c alcohol. The digital DNA-DNA hybridization and average nucleotide identity values between strain YIM B08730T and the reference strain S. isronensis JCM 13838T were 24.8 % and 81.2 %, respectively. The G + C content of the genomic DNA was 37.1 mol%. The genome of the novel strain contained genes associated with the production of siderophores, and it also revealed other functional gene clusters involved in plant growth promotion and soil bioremediation. Based on these phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM B08730T is considered to be a novel species of the genus Solibacillus, for which the name Solibacillus ferritrahens sp. nov. is proposed. The type strain is YIM B08730T (= NBRC 116268T = CGMCC 1.60169T).

Whole genome sequencing provides evidence for Bacillus velezensis SH-1471 as a beneficial rhizosphere bacterium in plants.

Bacillus is widely used in agriculture due to its diverse biological activities. We isolated a Bacillus velezensis SH-1471 from the rhizosphere soil of healthy tobacco, which has broad-spectrum antagonistic activity against a variety of plant pathogenic fungi such as Fusarium oxysporum, and can be colonized in the rhizosphere of a variety of plants. This study will further explore its mechanism by combining biological and molecular biology methods. SH-1471 contains a ring chromosome of 4,181,346 bp with a mean G + C content of 46.18%. We identified 14 homologous genes related to biosynthesis of resistant secondary metabolite, and three clusters encoded potential new antibacterial substances. It also contains a large number of genes from colonizing bacteria and genes related to plant bacterial interactions. It also contains genes related to environmental stress, as well as genes related to drug resistance. We also found that there are many metabolites in the strain that can inhibit the growth of pathogens. In addition, our indoor pot test found that SH-1471 has a good control effect on tomato wilt, and could significantly improve plant height, stem circumference, root length, root weight, and fresh weight and dry weight of the aboveground part of tomato seedlings. Therefore, SH-1471 is a potential biological control strain with important application value. The results of this study will help to further study the mechanism of SH-1471 in biological control of plant diseases and promote its application.

Reacting Molecular Oxygen with Butanone under Visible Light Irradiation: A General Aerobic Oxidation of Alkenes, Sulfides, Phosphines, and Silanes.

Here, we present a novel oxidation technique by reacting molecular oxygen with butanone under visible light irradiation. This method enables the mild oxidation of various functionalized compounds, including olefins, sulfides, phosphines, and silanes. Preliminary mechanistic experiments and theoretical calculations suggest that visible light triggers molecular oxygen to produce singlet oxygen in butanone. This singlet oxygen then reacts with butanone, producing an active oxidizing species.

[Construction of a 10rolGLP-1-expressing glucose-lowing Saccharomyces cerevisiae by CRISPR/Cas9 technique].

To develop a novel glucose-lowering biomedicine with potential benefits in the treatment of type 2 diabetes, we used the 10rolGLP-1 gene previously constructed in our laboratory and the CRISPR/Cas9 genome editing technique to create an engineered Saccharomyces cerevisiae strain. The gRNA expression vector pYES2-gRNA, the donor vector pNK1-L-PGK-10rolGLP-1-R and the Cas9 expression vector pGADT7-Cas9 were constructed and co-transformed into S. cerevisiae INVSc1 strain, with the PGK-10rolGLP-1 expressing unit specifically knocked in through homologous recombination. Finally, an S. cerevisiae strain highly expressing the 10rolGLP-1 with glucose-lowering activity was obtained. SDS-PAGE and Western blotting results confirmed that two recombinant strains of S. cerevisiae stably expressed the 10rolGLP-1 and exhibited the desired glucose-lowering property when orally administered to mice. Hypoglycemic experiment results showed that the recombinant hypoglycemic S. cerevisiae strain offered a highly hypoglycemic effect on the diabetic mouse model, and the blood glucose decline was adagio, which can avoid the dangerous consequences caused by rapid decline in blood glucose. Moreover, the body weight and other symptoms such as polyuria also improved significantly, indicating that the orally hypoglycemic S. cerevisiae strain that we constructed may develop into an effective, safe, economic, practical and ideal functional food for type 2 diabetes mellitus treatment.

[Health Benefit Evaluation for PM2.5 as Well as O3-8h Pollution Control in Chengdu, China from 2016 to 2020].

Both particulate matter with aerodynamics of less than 2.5 (PM2.5) and ozone are the two main air pollutants in China, which seriously endanger human health. To estimate the adverse impacts of PM2.5 and ozone on human health during the implementation of air pollution prevention and control actions in Chengdu, both the generalized additive model and the nonlinear distribution lag model of epidemiology were adopted to explore the exposure-response relationship coefficients ß of daily ozone 8h maximum concentration average (O3-8h), as well as that of PM2.5 on disease deaths in Chengdu from 2014 to 2016. On this basis, the environmental risk model and environmental value assessment model were both adopted to evaluate the health effects and health benefits in Chengdu from 2016 to 2020, respectively, with the assumption that PM2.5 and O3-8h concentration were reduced to specified air pollution control limits (35 µg·m-3 and 70 µg·m-3, respectively). The results showed 1 the annual concentration of PM2.5 presented gradually decreasing trends in Chengdu from 2016 to 2020. Specifically, ρ(PM2.5) from 63 µg·m-3 in 2016 decreased to 40.92 µg·m-3 in 2020. The average annual decline rate was approximately 9.8%. In contrast, the annual concentration of O3-8h from 155 µg·m-3 in 2016 increased to 169 µg·m-3 in 2020, and the increasing rate was approximately 2.4%. 2 Both PM2.5 and O3-8h had lag effects on three types of disease deaths. Under the maximum lag effect, the corresponding exposure-response relationship coefficients ß of PM2.5were 0.0003600, 0.0005001, and 0.0009237 for all-cause, cardiovascular, and respiratory premature deaths, respectively, whereas the corresponding ß of O3-8h were 0.0003103, 0.0006726, and 0.0007002, respectively. 3 If ρ(PM2.5) was reduced to the national secondary standard limit (35 µg·m-3), the corresponding number of health beneficiaries and economic benefits declined yearly. Specifically, the health beneficiary number of all-cause, cardiovascular, and respiratory disease deaths were reduced from 1128, 416, and 328 in 2016 to 229, 96, and 54 in 2020, respectively. There were a total number of 3314 avoidable premature deaths for all-cause diseases during the five years, resulting in a total health economic benefit of 7.66 billion yuan. 4 If we assume that ρ(O3-8h) was reduced to the concentration limit specified by the World Health Organization (70 µg·m-3), the corresponding number of health beneficiaries and economic benefits were increasing yearly. Specifically, the health beneficiaries' numbers of all-cause, cardiovascular, and respiratory disease deaths rose from 1919, 779, and 606 in 2016 to 2429, 1157, and 635 in 2020, respectively. The annual average growth rates of avoidable all-cause and cardiovascular mortality were 6.85% and 10.72%, respectively, which was higher than the annual average rise rate of ρ(O3-8h). There were 10790 total avoidable deaths from all-cause diseases during the five years, resulting in a total health economic benefit of 26.62 billion yuan. These findings indicate that PM2.5 pollution in Chengdu had been well controlled, whereas O3 pollution had become more severe and had become another key air pollutant threatening human health. Therefore, the synchronous control of PM2.5 and ozone should be implemented in the future.

Differential Responses of Bacterial and Fungal Communities to Siderophore Supplementation in Soil Affected by Tobacco Bacterial Wilt (Ralstonia solanacearum).

Siderophores secreted by microorganisms can promote ecological efficiency and could be used to regulate the unbalanced microbial community structure. The influence of the siderophore activity of Trichoderma yunnanense strain 2-14F2 and Beauveria pseudobassiana strain (2-8F2) on the physiological/biochemical functions and community structure of soil microbes affected by tobacco bacterial wilt (TBW) was studied. DNS Colorimetry and Biolog-eco plates were used to quantify the impacts of strain siderophores on soil enzyme activities and microbial metabolism. Based on Illumina MiSeq high-throughput sequencing, the soil 16S rDNA and ITS sequences were amplified to dissect the response characteristics of alpha/beta diversity and the structure/composition of a soil microbial community toward siderophores. The KEGG database was used to perform the PICRUSt functional prediction of the microbial community. We found that siderophores of 2-14F2 and 2-8F2, at certain concentrations, significantly increased the activities of sucrase (S-SC) and urease (S-UE) in the TBW soil and enhanced the average well color development (AWCD, carbon source utilization capacity) of the microbial community. The metabolic capacity of the diseased soil to amino acids, carbohydrates, polymers, aromatics, and carboxylic acids also increased significantly. The response of the bacterial community to siderophore active metabolites was more significant in alpha diversity, while the beta diversity of the fungal community responded more positively to siderophores. The relative abundance of Actinobacteria, Chloroflexi, and Acidobacteria increased and was accompanied by reductions in Proteobacteria and Firmicutes. LEfSe analysis showed that Pseudonocardiaceae, Gemmatimonas, Castellaniella, Chloridiumand and Acrophialophora altered the most under different concentrations of siderophore active metabolites. The PICRUSt functional prediction results showed that siderophore increased the abundance of the redox-related enzymes of the microbial community in TBW soil. The BugBase phenotypic prediction results showed that the siderophore activity could decrease the abundance of pathogenic bacteria. The study concludes that siderophore activity could decrease the abundance of pathogenic bacteria and regulate the composition of the microbial community in TBW soil. The activities of sucrase (S-SC) and urease (S-UE) in TBW soil were significantly increased. Overall, the siderophore regulation of community structures is a sustainable management strategy for soil ecosystems.

Chitinolyticbacter albus sp. Nov., A Novel Chitin-Degrading Bacterium Isolated from Ancient Wood Rhizosphere Soil.

In this study, a novel aerobic mesophilic bacterial strain with capable of degrading chitin, designated YIM B06366T, was isolated and classified. The rod-shaped, Gram-stain-negative, on-spore-forming bacterium originated from rhizosphere soil sample collected in Kunming City, Yunnan Province, southwest PR China. Strain YIM B06366T exhibited growth at temperatures between 20 and 35 °C (optimum, 30 °C) and at pH 6.0-8.0 (optimum, pH 6.0). The analysis of 16S rRNA gene sequence similarity revealed that strain YIM B06366T was most closely related to type strain Chitinolyticbacter meiyuanensis SYBC-H1T (98.9%). Phylogenetic analysis based on genome data indicated that strain YIM B06366T should be assigned to the genus Chitinolyticbacter. The Average Nucleotide Identity (ANI) and digital DNA-DNA Hybridization (dDDH) values between strain YIM B06366T and the reference strain Chitinolyticbacter meiyuanensis SYBC-H1T were 84.4% and 27.7%, respectively. The major fatty acids included Summed Feature 3 (C16:1 ω6c/C16:1 ω7c), Summed Feature 8 (C18:1 ω6c/C18:1 ω7c), and C16:0. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, aminophospholipids, and two unidentified phospholipids. The predominant menaquinone was Q-8, and the genomic DNA G + C content was 64.1%. Considering the polyphasic taxonomic evidence, strain YIM B06366T is proposed as a novel species within the genus Chitinolyticbacter, named Chitinolyticbacter albus sp. nov. (type strain YIM B06366T = KCTC 92434T = CCTCC AB 2022163T).

Pseudomonas subflava sp. nov., a new Gram-negative bacterium isolated from Guishan in Yunnan province, south-west China.

A new Gram-negative, rod-shaped, flagellated bacterium was isolated from soil in the Guishan, Xinping County, Yuxi City, Yunnan Province, China, and named YIM B01952T. Growth occurred at 10-40 °C (optimum, 30 °C), pH 6.0-9.0 (optimum, pH 7.5) and with up to ≤ 5.0% (w/v) NaCl on Tryptic Soy Broth Agar (TSA) plates. Phylogenetic analysis based on the 16S rRNA gene and draft-genome sequence showed that strain YIM B01952T belonged to the genus Pseudomonas, and was closely related to the type strain of Pseudomonas alcaligenes (sequence similarity was 98.8%). The digital DNA-DNA hybridization (dDDH) value between strain YIM B01952T and the parallel strain P. alcaligenes ATCC 14909T was 49.0% based on the draft genome sequence. The predominant menaquinone was Q-9. The major fatty acids were summed feature 8 (C18:1 ω6c and/or C18:1 ω7c), summed feature 3 (C16:1 ω6c and/or C16:1 ω7c) and C16:0. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol. The genome size of strain YIM B01952T was 4.341 Mb, comprising 4156 predicted genes with a DNA G + C content of 66.4 mol%. In addition, we detected that strain YIM B01952T had some traditional functional genes (plant growth promotion and multidrug resistance), unique genes through genome comparison and analysis with similar strains. Based on genetic analyses and biochemical characterization, the strain YIM B01952T was identified as a novel species in the genus Pseudomonas, for which the name Pseudomonas subflava sp. nov. is proposed. The type strain is YIM B01952T (=CCTCC AB 2021498T = KCTC 92073T).

Metal Coordinated Polymer as Three-Dimensional Network Binder for High Sulfur Loading Cathode of Lithium-Sulfur Battery.

The construction of high sulfur (S) loading cathode is one of the critical parameters to obtain lithium-sulfur (Li-S) batteries with high energy density, but the slow redox reaction rate of high S loading cathode limits the development process. In this paper, a metal coordinated polymer-based three-dimensional network binder, which can improve the reaction rate and stability of S electrode. Compared with traditional linear polymer binders, the metal coordinated polymer binder can not only increase the load amount of S through the three-dimensional cross-linking, but also promote the interconversion reactions between S and lithium sulfide (Li2 S), avoiding the passivation of electrode and improving the stability of the positive electrode. At an S load of 4-5 mg cm-2 and an E/S ratio of 5.5 µL mg-1 , the discharged voltage in the second platform is 2.04 V and the initial capacity is 938 mA h g-1 with metal coordinated polymer binder. Moreover, the capacity retention rate approaches 87% after 100 cycles. In comparison, the discharged voltage in the second platform is lost and the initial capacity is 347 mA h g-1 with PVDF binder. It demonstrates the advanced properties of metal-coordinated polymer binders for improving the performance of Li-S batteries.

Brevibacillus daliensis sp. nov., Isolated From Soil in Machangqing Nature Reserve.

A new aerobic bacterial strain, designated strain YIM B02290T, was isolated from the soil of Machangqing, Dali city, Yunnan Province, China. Cells were Gram-stain-positive, sporogenous, rod-shaped, and motile with peritrichous flagella. Strain YIM B02290T showed the highest 16S rRNA gene sequence similarity with Brevibacillus laterosporus (97.6%) and Brevibacillus halotolerans (97.6%). The ANI and dDDH values between strain YIM B02290T and the two reference strains Brevibacillus laterosporus LAM00312T and Brevibacillus halotolerans DSM 25T are 72.6% and 72.2%, 20.2% and 19.5% based on the draft genome sequence, respectively. The major cellular fatty acids contain anteiso-C15: 0 and iso-C15: 0. The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid. The predominant menaquinone was identified as menaquinone-7. The main polar lipids of strain YIM B02290T were diphosphatidylglycerol, phosphatidylglycerol, phospholipid, phosphatidyl monomethylethanolamine. The genomic DNA G + C content was 40.6 mol%. All results showed that strain YIM B02290T represents a novel species of the genus Brevibacillus, for which the name Brevibacillus daliensis sp. nov. is proposed. The type strain is YIM B02290T (= CCTCC AB 2021094T = CGMCC 1.18802T = KCTC 43376T).

Enhanced Separation Performance of Hierarchically Porous Membranes Fabricated via the Combination of Crystallization Template and Foaming.

In this work, poly (vinylidene fluoride) (PVDF) hierarchically porous membranes (HPMs) with isolated large pores and continuous narrow nano-pores have been fabricated from its blend with poly (methyl methacrylate) (PMMA) based on the combination of crystallization template with chemical or supercritical CO2 foaming. On the one hand, the decomposition of azodicarbonamide (ADC, chemical foaming agent) or the release of CO2 can produce isolated large pores. On the other hand, PMMA is expelled during the isothermal crystallization of PVDF in their miscible blend, yielding narrow nano-pores upon etching with a selective solvent. In the case of supercritical CO2, the attained PVDF HPMs fail to improve separation performance because of the compact wall of isolated-large-pore and consequent poor connectivity of hierarchical pores. In the case of ADC, the optimal HPM exhibits much higher flux (up to 20 times) without any loss of selectivity compared with the reference only with nano-pores. The enhanced permeability can be attributed to the shorter diffusion length and lower diffusion barrier from isolated large pores, while the comparable selectivity is determined by narrow nano-pores in THE matrix.

Cohnella mopanensis sp. nov., a new Gram-negative bacterium isolated from soil in Xinping Mountain National Forest Park.

A Gram-stain-negative, aerobic, rod-shaped bacteria strain, named YIM B01951T, was isolated from a forest soil sample collected from Mopan Mountain National Forest Park, Xinping City, Yunnan Province, southwest PR China (101°58'06" N, 23°03'02" E). Growth occurred at 15-40 °C (optimum, 30 °C), pH 5.0-8.0 (optimum, pH 6.5) and with up to ≤ 3.0% (w/v) NaCl on Nutrient Agar plates. The results of 16S rRNA gene sequence similarity analysis showed that strain YIM B01951T was closely related to the type strain of Cohnella arctica M9-62T (96.5%) and Cohnella lupini RLAHU4BT (96.3%). YIM B01951T contains anteiso-C15:0 and iso-C16:0 as the major cellular fatty acids; the main polar lipids are diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), lysylphosphatidylglycerol (PGL) and five aminophospholipids (APL). The MK-7 is the major respiratory quinone and the DNA G + C content is 49.2 mol%. Based on these phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM B01951T is considered to be a novel species of the genus Cohnella, and named Cohnella mopanensis sp. nov. The type strain is YIM B01951T (= NBRC 115331T = KCTC 43370T).

Exendin-4 may improve type 2 diabetes by modulating the epigenetic modifications of pancreatic histone H3 in STZ-induced diabetic C57BL/6 J mice.

Type 2 diabetes (T2D) is a complicated systemic disease that might be improved by exendin-4, although the epigenetic role remains unclear. In the current study, C57BL/6 J mice were used to generate a T2D model, followed by treatment with exendin-4 (10 µg/kg). Histone H3K9 and H3K23 acetylation, H3K4 mono-methylation, and H3K9 di-methylation were explored by western blot analysis of pancreatic histone extracts. Real-time polymerase chain reaction (PCR) was used to examine the expression levels of pancreatic beta cell development-related genes, and chromatin immunoprecipitation (ChIP) was applied to analyze H3 and H3K9 acetylation, H3K4 mono-methylation, and H3K9 di-methylation in the promoter region of the pancreatic and duodenal homeobox 1 (Pdx1) gene. The results showed that total H3K9 di-methylation and H3K9 and H3K23 acetylation increased in pancreatic tissues of diabetic mice, whereas H3K4 mono-methylation was reduced. All of these changes could be abrogated by treatment with exendin-4. Our data indicated that T2D progression might be improved by exendin-4 treatment through the reversal of global pancreatic histone H3K9 and H3K23 acetylation, H3K4 mono-methylation, and H3K9 di-methylation. A better understanding of these epigenetic alterations may, therefore, lead to novel therapeutic strategies for T2D.

Propioniciclava soli sp. nov., isolated from forest soil, Yunnan, China, and reclassification of the genus Brevilactibacter into the genus Propioniciclava, and Brevilactibacter sinopodophylli, Brevilactibacter flavus, and Brevilactibacter coleopterorum as Propioniciclava sinopodophylli comb. nov., Propioniciclava flava comb. nov., and Propioniciclava coleopterorum comb. nov., respectively.

A Gram-stain-positive, coccus-shaped, facultatively anaerobic, non-motile bacterial strain, designated YIM S02567T, was isolated from a forest soil sample collected from Gejiu City, Yunnan Province, southwest PR China. Growth was observed at 10-45 °C, at pH 6.0-9.5, in the presence of up to 4.0% (w/v) NaCl on R2A medium. The results of 16S rRNA gene sequence similarity analysis showed that strain YIM S02567T was most closely related to the type strain of Brevilactibacter sinopodophylli (95.4%) and Propioniciclava tarda (94.7%), and phylogenetic analysis based on genome data showed that strain YIM S02567T should be assigned to the genus Propioniciclava. The cell-wall diamino acid was meso-diaminopimelic acid. The major cellular fatty acids were identified as anteiso-C15:0 and C16:0, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and two unidentified glycolipids. The predominant menaquinone was MK-9(H4). The genomic DNA G + C content was 71.2 mol%. Based on the polyphasic taxonomic evidence, strain YIM S02567T is assigned to a novel member of the genus Propioniciclava, for which the name Propioniciclava soli sp. nov., (type strain YIM S02567T = CCTCC AB 2020128T = CGMCC 1.18504T = KCTC 49478T) is proposed. Furthermore, we propose the reclassification of Brevilactibacter as Propioniciclava gen. nov.

Viridibacillus soli sp. nov., isolated from forest soil in Ailaoshan National Nature Reserve.

A Gram stain-positive, rod-shaped, and subterminal endospore-forming bacterium, designated strain YIM B01967T, was isolated from a forest soil sample collected in Ailaoshan National Nature Reserve, Yuxi City, Xinpin county, Yunnan province, China. Strain YIM B01967T showed the highest 16S rRNA gene sequence similarity with Viridibacillus arvi (99.1%) and Viridibacillus arenosi (98.9%). Based on the phylogenetic and 16S rRNA gene sequence results, strain YIM B01967T was affiliated to the genus Viridibacillus. The growth of YIM B01967T was observed at 15-35 °C (optimum, 28 °C), pH 7.0-9.0 (optimum, pH 7.5) and in the presence of 0-2% (w/v) NaCl (optimum in 2% NaCl). The cell wall sugars include ribose, glucose, arabinose, galactose, and mannose. The quinone system consisted of the major compound MK-8 and moderate amounts of MK-7. The major fatty acids (> 10%) included iso-C15:0, anteiso-C15:0, C16:1 ω10c. The major polar lipids profile included DPG, PME. The cell wall peptidoglycan was most likely of the type A4α with an L-Lys-D-Asp interpeptide bridge. The genomic DNA G + C content of strain YIM B01967T was 36.3 mol%. The ANI and digital DNA-DNA hybridization (dDDH) values between strain YIM B01967T and Viridibacillus arvi DSM 16317 T, Viridibacillus arenosi DSM 16319 T were 61.0% and 32.1%, 60.0% and 33.1% based on the draft genome sequence. The results support the conclusion that strain YIM B01967T represents a novel species of the genus Viridibacillus, for which the name Viridibacillus soli sp. nov., is proposed. The type strain is YIM B01967T (= KCTC 43249 T = CGMCC 1.18436 T).

Optimized biosynthesis of santalenes and santalols in Saccharomyces cerevisiae.

Santalenes and santalols from Santalum album are the main components of the valuable spice sandalwood essential oil, which also has excellent pharmacological activities such as antibacterial, anti-inflammatory, and antitumor. Firstly, we constructed biosynthesis pathways of santalenes by synthetic biology strategy. The assembled biosynthetic cassettes were integrated into the multiple copy loci of δ gene in S. cerevisiae BY4742 with assistance of pDi-CRISPR, and 94.6 mg/L santalenes was obtained by shake flask fermentation of engineered yeast. Secondly, a selected optimized P450-CPR redox system was integrated into the chromosome of the santalenes-producing strain with a single copy, and 24.6 mg/L santalols were obtained. Finally, the yields of santalenes and santalols were increased to 164.7 and 68.8 mg/L, respectively, by downregulating ERG9 gene. This is the first report on the de novo synthesis of santalols by P450-CPR chimera in S. cerevisiae. Meanwhile, the optimized chimeric CYP736A167opt-46tATR1opt exhibits higher activity to oxidize santalenes into santalols. It would provide a feasible solution for the optimal biosynthesis of santalols. KEY POINTS: • First-time de novo synthesis of santalols by P450-CPR chimera in S. cerevisiae. • Truncated 46tATR1 has higher activity than that of CPR2. • Yields of santalenes and santalols were increased by downregulating ERG9 gene.

Functional Characterization of a New Bifunctional Terpene Synthase LpNES1 from a Medicinal Plant Laggera pter odonta.

Laggera pterodonta, known in China as 'Choulingdan' for its stimulous odor, has long been used as traditional herbal medicine. The essential oil of L. pterodonta, which exhibits various pharmacological activities, is a rich resource of monoterpenes and sesquiterpenes. To date, however, the terpene synthases responsible for their production remain unknown. In present study, a new terpene synthase gene (LpNES1) was identified from L. pterodonta, transcript level of which was significantly upregulated in response to methyl jasmonate treatment. Recombinant LpNES1 could synthesize (E)-nerolidol and minor ß-farnesene from farnesyl diphosphate and linalool from geranyl diphosphate in vitro. Whereas, only sesquiterpenes including (E)-nerolidol and minor ß-farnesene were released when LpNES1 was reconstituted in yeast, even coexpressed with a geranyl diphosphate synthase (ERG20WW). Combined with subcellular localization experiment, the result indicated that the cytosol-targeted LpNES1 was responsible for (E)-nerolidol biosynthesis exclusively in L. pterodonta. Additionally, the expression level of LpNES1 gene was more prominent in floral buds than that in other tissues. LpNES1 characterized in present study not only lays the molecular foundation for sesquiterpene biosynthesis of L. pterodonta, but provides a key element for further biosynthesis of bioactive compound in microbes.

Aestuariimicrobium ganziense sp. nov., a new Gram-positive bacterium isolated from soil in the Ganzi Tibetan autonomous prefecture, China.

A novel Gram-stain positive, oval-shaped, and non-flagellated bacterium, designated YIM S02566T, was isolated from alpine soil in Shadui Towns, Ganzi County, Ganzi Tibetan Autonomous Prefecture, Sichuan Province, PR China. Growth occurred at 23-35 °C (optimum, 30 °C) in the presence of 0.5-4% (w/v) NaCl (optimum, 1%) and at pH 7.0-8.0 (optimum, pH 7.0). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain YIM S02566T was most closely related to the genus Aestuariimicrobium, with Aestuariimicrobium kwangyangense R27T and Aestuariimicrobium soli D6T as its closest relative (sequence similarities were 96.3% and 95.4%, respectively). YIM S02566T contained LL-diaminopimelic acid in the cell wall. MK-9(H4) was the predominant menaquinone. The major fatty acid patterns were anteiso-C15:0 (60.0%). The major polar lipid was DPG. The genome size of strain YIM S02566T was 3.1 Mb, comprising 3078 predicted genes with a DNA G + C content of 69.0 mol%. Based on these genotypic, chemotaxonomic and phenotypic evidences, strain YIM S02566T was identified as a novel species in the genus Aestuariimicrobium, for which the name Aestuariimicrobium ganziense sp. nov. is proposed. The type strain is YIM S02566T (= CGMCC 1.18751 T = KCTC 49,477 T).