摘要：
Objectives: Human infection caused by an uncommon Salmonella enterica subsp. diarizonae (hereafter S. diarizonae) is rising. However, knowledge concerning S. diarizonae is still limited. This study aimed to investigate the genomic features of S. diarizonae S499 isolated from a child patient with gastroenteritis symptom in China. Methods: The antimicrobial susceptibility of S. diarizonae S499 was determined by microdilution broth assay. Whole genome was sequenced using Illumina HiSeq X-10 and PacBio RS II platforms and was de novo assembled using Unicycler and SPAdes. Conjugation experiment was performed by a broth mating method. Results: S. diarizonae S499 was a multi-drug resistant (MDR) isolate and showed resistance to all cephalosporin drugs tested. Six plasmids (pS0499A, pS0499B, pS0499C, pS0499D, pS0499E and pS0499F) were identified. A rare gene cassette IS26-bla(CTX-M-55)-wbuc-Delta bla(TEM-1) -IS26-intI1 was repeatedly inserted into pS0499A three times in one locus and reversely inserted into plasmid pS0499D. That enhanced cephalosporin resistance. To the best of our knowledge, this finding has not been reported previously. Both pS0499A and pS0499B contained multiple resistance genes and could transfer to recipient strain E. coli EC600. Conclusion: This article reported the genome features of S. diarizonae S499, which contained four resistant plasmids including a novel plasmid pS0499A with a novel gene cassette rearrangement. These data could contribute to a better understanding of the antimicrobial resistance mechanisms and transmission dynamics of S. diarizonae. (c) 2022 The Author(s). Published by Elsevier Ltd on behalf of International Society for Antimicrobial Chemotherapy. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
期刊：
International Journal of Food Properties,2022年25(1):1203-1214 ISSN：1094-2912
通讯作者：
Shoulei Yan
作者机构：
[Li, Jie; Yan, Shoulei; Liu, Yanzhao; Wang, Jie] Huazhong Agr Univ, Coll Food Sci & Technol, Wuhan 430070, Peoples R China.;[Li, Jie; Yan, Shoulei] Yangtze River Econ Belt Engn Res Ctr Green Dev Bu, Minist Educ, Wuhan, Peoples R China.;[Li, Jie; Yan, Shoulei] Hubei Aquat Vegetable Preservat Proc Engn Technol, Wuhan, Peoples R China.;[Diao, Ying] Wuhan Polytech Univ, Coll Life Sci & Technol, Wuhan, Peoples R China.;[Hu, Zhongli] Wuhan Univ, Coll Life Sci, Hubei Lotus Rhizome Engn Technol Res Ctr, Wuhan, Peoples R China.
通讯机构：
[Shoulei Yan] C;College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China<&wdkj&>Yangtze River Economic Belt Engineering Research Center for Green Development of Bulk Aquatic Bioproducts Industry of Ministry of Education, Wuhan, China<&wdkj&>Hubei Aquatic Vegetable Preservation Processing Engineering Technology Research Center, Wuhan, China
关键词：
Lotus rhizome starch;Adulteration;Ferrous sulfate;polyphenol;Liquid chromatography mass spectrometry;Colorimetric card
摘要：
Lotus rhizome starch (LRS) is an important traditional processed product of lotus rhizome in China due to its good taste and high nutritional value. However, adulterated lotus rhizome starches are prevalent on the market. In this study, a novel method for the rapid detection of adulteration in LRS based on color reaction between ferrous sulfate (FS) and (-)-gallocatechin (GC) in LRS was studied. The polyphenols in LRS were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS) as GC and (+)-catechin, and GC could show a bluish color with FS solution. LRS showed a bluish color when FS solution (0.01 mol/L) was used as the chromogenic agent, which was not observed for other ten kinds of starches tested in this work. There was an obvious color gradient when FS solution was used to react with adulterated LRS at different proportions, and the change in blue color was obvious when maize starch or cassava starch was adulterated in LRS at the ratio of 6:4. Then, a semi-quantitative method was used to make a color card according to the color gradient, which was then successfully used to detect the adulteration of four kinds of commercial LRS. The results indicate that the proposed method may facilitate the simple, effective and efficient detection of adulteration in LRS.
摘要：
Pseudonocardia species are emerging as important microorganisms of global concern with unique and increasingly significant ecological roles and represent a prominent source of bioactive natural products, but genetic engineering of these organisms for biotechnological applications is greatly hindered due to the limitation of efficient genetic manipulation tools. In this regard, we report here the establishment of an efficient genetic manipulation system for a newly isolated strain, Pseudonocardia alni Shahu, based on plasmid conjugal transfer from Escherichia coli to Pseudonocardia. Conjugants were yielded upon determining the optimal ratio between the donor and recipient cells, and designed genome modifications were efficiently accomplished, including exogenous gene integration based on an integrative plasmid and chromosomal stretch removal by homologous recombination using a suicidal non-replicating vector. Collectively, this work has made the P. alni Shahu accessible for genetic engineering, and provided an important reference for developing genetic manipulation methods in other rare actinomycetes.
摘要：
Cardiovascular complications of patients with type 2 diabetes mellitus (T2DM) threaten the health and life of numerous individuals. Recently, growth factor receptor-binding protein 10 (GRB10) was found to play a pivotal role in vascular complications of T2DM, which participates in the regulation of lipid metabolism of T2DM patients. The genetic variation of GRB10 rs1800504 is closely related to the risk of coronary heart disease in patients with T2DM. The development of GRB10 as a key mediator in the association of lipid metabolism with cardiovascular complications in T2DM is detailed in and may provide new potential concerns for the study of cardiovascular complications in T2DM patients.
关键词：
Auto-induction;Escherichia coli expression;glycoside hydrolase family 5;mannanase;mannooligosaccharides;Pichia pastoris expression.
摘要：
Background: Mannans are the main components of hemicellulose in nature and serve as the major storage polysaccharide in legume seeds. To mine new mannanase genes and identify their functional characteristics are an important basis for mannan biotechnological applications.<&wdkj&>Objective: In this study, a putative mannanase gene (ManBs31) from the genome of the marine bacterium Alteromonadaceae Bs31 was characterized.<&wdkj&>Methods: Amino acid sequence analysis and protein structural modeling were used to reveal the molecular features of ManBs31. The catalytic domain of ManBs31 was recombinantly produced using Escherichia coli and Pichia pastoris expression systems. The biochemical properties of the enzymes were determined by reducing sugar assay and thin-layer chromatography.<&wdkj&>Results: Sequence analysis revealed that ManBs31 was a multidomain protein, consisting of a catalytic domain belonging to glycoside hydrolase family 5 (GH5) and two cellulose-binding domains. Recombinant ManBs31-GH5 exhibited the maximum hydrolytic performance at 70 ºC and pH 6. It showed the best hydrolysis capacity toward konjac glucomannan (specific enzyme activity up to 1070.84 U/mg) and poor hydrolysis ability toward galactomannan with high side-chain modifications (with a specific activity of 344.97 U/mg and 93.84 U/mg to locust bean gum and ivory nut mannan, respectively). The hydrolysis products of ManBs31-GH5 were mannooligosaccharides, and no monosaccharide was generated. Structural analysis suggested that ManBs31-GH5 had a noncanonical +2 subsite compared with other GH5 mannanases.<&wdkj&>Conclusion: ManBs31 was a novel thermophilic endo-mannanase and it provided a new alternative for the biodegradation of mannans, especially for preparation of probiotic mannooligosaccharides.
通讯机构：
[Xiaonan Liu; Huifeng Jiang] K;Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China<&wdkj&>University of Chinese Academy of Sciences, Beijing, 100049, China<&wdkj&>National Center of Technology Innovation for Synthetic Biology, Tianjin, 300308, China
关键词：
Scutellarin;Yarrowia lipolytica;Metabolic engineering;Combinatorial gene overexpression;Peer review under responsibility of KeAi Communications Co.,Ltd
摘要：
Scutellarin related drugs have superior therapeutic effects on cerebrovascular and cardiovascular diseases.Here,an optimal biosynthetic pathway for scutellarin was constructed in Yarrowia lipolytica platform due to its excellent metabolic potential.By integrating multi-copies of core genes from different species,the production of scutellarin was increased from 15.11 mg/L to 94.79 mg/L and the ratio of scutellarin to the main by-product was improved about 110-fold in flask condition.Finally,the production of scutellarin was improved 23-fold and reached to 346 mg/L in fed-batch bioreactor,which was the highest reported titer for de novo production of scutellarin in microbes.Our results represent a solid basis for further production of natural products on unconventional yeasts and have a potential of industrial implementation.
摘要：
Trapa bispinosa Roxb. is a traditional Chinese food which is well known for its medicinal properties. The shell of Trapa bispinosa has anticancer activity, maybe due to its high content of polyphenols. There are few studies on the chemical composition of Trapa bispinosa shells, then we isolated the active components from Trapa bispinosa shell and clarified the mechanism of its anticancer activity. One monomer compound was separated from the ethanol extract of the Trapa bispinosa shell by fractional extraction, silica gel, Sephadex LH-20 gel column chromatography and liquid phase separation. The structure, identified by NMR was 1,2,3,6-tetra-O-galloyl-β-D-glucose. The results of the CCK-8 assay showed that 1,2,3,6-tetra-O-galloyl-β-D-glucose could significantly inhibit the proliferation of gastric cancer SGC7901 cells, and the effect was close to that of 5-fluorouracil. Here, 1,2,3,6-tetra-O-galloyl-β-D-glucose could affect the cell cycle of SGC7901 cells. At the dose of 200 μg/mL and an incubation time of 48 h, SGC7901 cells remained in the G1 phase, apoptosis occurred, the intracellular calcium ion concentration increased and the mitochondrial membrane potential decreased. Transcriptome sequencing analysis showed that the differentially expressed genes were mainly enriched in the P53 signalling pathway associated with apoptosis. The results of qPCR and Western blot showed that 1,2,3,6-tetra-O-galloyl-β-D-glucose could induce apoptosis of SGC7901 cells by up-regulating the expression levels of P21, PUMA, PERP and IGF-BP3 genes, down-regulating the CyclinD gene, increasing the expression levels of cytochrome C, caspase-3, caspase-9 protein and decreasing that of the protein BCL-2.
作者机构：
[Chen, Hongbiao; Li, Yun; Cao, Hui; Zhao, Xiuju] Wuhan Polytech Univ, Sch Biol & Pharmaceut Engn, NDHN, Team Neonatal & Infant Dev Hlth & Nutr, Wuhan 430023, Peoples R China.;[Li, Yun; Yi, Ping] Kindstar Global Precis Med Inst, Wuhan 430223, Peoples R China.;[Wang, Qi] Wuhan Polytech Univ, Sch Food Engn, Wuhan 430023, Peoples R China.
通讯机构：
[Ping Yi; Xiuju Zhao] A;Authors to whom correspondence should be addressed.<&wdkj&>Team of Neonatal & Infant Development, Health and Nutrition, NDHN, School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China<&wdkj&>Authors to whom correspondence should be addressed.<&wdkj&>Kindstar Global Precision Medicine Institute, Wuhan 430223, China
摘要：
The number of metabolic syndromes (MetS) is increasing, and a fish phospholipid diet can reduce the risk of MetS. In this study, the changes in lipid metabolism of colon contents were analyzed by extensive lipidomics in mice with metabolic syndrome by fish phospholipid diet, and mice were randomly divided into experimental groups with different diet types by establishing a MetS model. After 14 weeks, the mice were sacrificed and the serum and colon contents were collected. Ultra-high liquid phase tandem mass spectrometry was used for broadly targeted lipidomic analysis, and the qualitative and quantitative detection of lipid metabolism changes in the colonic contents of mice. Under the intervention of fish phospholipids, MetS mice were significantly inhibited, serum total cholesterol (TC) and triglycerides (TG) decreased, serum high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) levels were improved, fasting blood glucose and insulin levels decreased, and inflammatory factors decreased. Through screening, it was found that thirty-three lipid metabolites may be key metabolites and five have significantly changed metabolic pathways. Modularizing lipid metabolites, it is possible to understand the extent to which different types and concentrations of fish phospholipids affect metabolic syndrome. Therefore, our study may provide new therapeutic clues for improving MetS.
作者：
Ye, Shijie;Yin, Dongjie;Sun, Xiaoyan;Chen, Qinyi;Min, Ting;...
期刊：
Molecules,2022年27(23):8374- ISSN：1420-3049
通讯作者：
Limei Wang
作者机构：
[Wang, Hongxun; Chen, Qinyi; Ye, Shijie; Sun, Xiaoyan; Yin, Dongjie; Wang, Limei] Wuhan Polytech Univ, Coll Life Sci & Technol, Wuhan 430023, Peoples R China.;[Min, Ting] Wuhan Polytech Univ, Coll Food Sci & Engn, Wuhan 430023, Peoples R China.
通讯机构：
[Limei Wang] C;College of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, China<&wdkj&>Author to whom correspondence should be addressed.
摘要：
Trapa bispinosa Roxb. is an economical crop for medicine and food. Its roots, stems, leaves, and pulp have medicinal applications, and its shell is rich in active ingredients and is considered to have a high medicinal value. One of the main functional components of the Trapa bispinosa Roxb. shell is 1-galloyl-beta-D-glucose (beta G), which can be used in medical treatment and is also an essential substrate for synthesizing the anticancer drug beta-penta-o-Galloyl-glucosen (PGG). Furthermore, gallate 1-beta-glucosyltransferase (EC 2.4.1.136) has been found to catalyze gallic acid (GA) and uridine diphosphate glucose (UDPG) to synthesize beta G. In our previous study, significant differences in beta G content were observed in different tissues of Trapa bispinosa Roxb. In this study, Trapa bispinosa Roxb. was used to clone 1500 bp of the UGGT gene, which was named TbUGGT, to encode 499 amino acids. According to the specificity of the endogenous expression of foreign genes in Escherichia coli, the adaptation codon of the cloned original genes was optimized for improved expression. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbUGGT with squalene synthases from other plants. The TbUGGT gene was constructed into a PET-28a expression vector and then transferred into Escherichia coli Transsetta (DE3) for expression. The recombinant protein had a molecular weight of 55 kDa and was detected using SDS-PAGE. The proteins were purified using multiple fermentation cultures to simulate the intracellular environment, and a substrate was added for in vitro reaction. After the enzymatic reaction, the levels of beta G in the product were analyzed using HPLC and LC-MS, indicating the catalytic activity of TbUGGT. The cloning and functional analysis of TbUGGT may lay the foundation for further study on the complete synthesis of beta G in E. coli.
通讯机构：
[Li, R.; Zhou, M.] C;College of Biological and Pharmaceutical Engineering, China;College of Food Science and Engineering, 6 Garden South Road, 3-2-502, China
摘要：
Proteus mirabilis is an opportunistic human pathogen. In this study, a novel SXT/R391 integrative and conjugative element (ICE), named ICEPmiChnS012, was identified in the multidrug-resistant P. mirabilis strain S012 that was isolated from retail chicken in China. Whole genome sequencing revealed that ICEPmiChnS012 carried 22 resistance genes including aac(6′)-Ib-cr, fosA3, blaOXA-1, blaCTX-M-65, and blaHMS-1. ICEPmiChnS012 harbored 10 copies of IS26 and IS26-mediated genetic new rearrangements caused variations in HS4 region. To our knowledge, an unusual gene cassette array dfrA1-ereA1-aadA2 was found in P. mirabilis in this study for the first time. And this is the first report of identification of aph3-VI and blaHMS-1 in VRIII region in P. mirabilis. The conjugation experiments proved that ICEPmiChnS012 could be transferred to Escherichia coli EC600 through conjugation. These findings demonstrated that ICEPmiChnS012 was a special ICE that carried the largest number of antimicrobial resistance genes in the family of SXT/R391 ICEs. This element could serve as an important vehicle for the dissemination of antibiotic resistance genes and should receive great concern.
摘要：
Oxidative stress and inflammation in kidney are the main causes for hyperuricemic nephropathy (HN). Baicalin and baicalein, two flavonoids, have anti-inflammatory and anti-oxidative effects and they are interconvertible in the body. In this study, both baicalin and baicalein were administered by intragastric administration (i.g.) or intraperitoneal injection (i.p.) at the dose of 50 mg kg(-1), once a day for 15 consecutive days to HN mice, a model established by i.g. of yeast extract combined with i.p. of potassium oxonate. In HN mice, baicalin and baicalein reduced serum uric acid (SUA) levels and protected kidneys by anti-inflammatory and anti-oxidative effects. Mechanistically, the effect of baicalin and baicalein on reducing SUA levels might due to their inhibitory effect on xanthine oxidase (XO) activity in vivo and in vitro. Furthermore, the mechanisms of baicalin and baicalein against HN were analyzed with network pharmacology and molecular docking technology. The network pharmacology indicated that the protective effects of baicalin and baicalein against HN were mainly related to their down-regulating effects on TLRs, NF-κB, MAPK, PI3K/AKT and NOD-like receptor signaling pathways. Molecular docking indicated high binding affinity of baicalin/baicalein to targets such as AKT1 and MAPK1. In summary, baicalin and baicalein are promising drug candidates for the treatment of HN by inhibiting XO activity, reducing inflammation and cell apoptosis through down-regulating TLRs/NLRP3/NF-κB, MAPK, PI3K/AKT/NF-κB pathways.
摘要：
Lotus root (Nelumbo nucifera G.) is a high economic value crop in the world. In this study, the storage characteristics (color, sensory, texture, and fatty acids) of lotus root ("Elian No.5") were evaluated at different harvest periods (September 2018, October 2018, November 2018, December 2018, and January 2019). Moreover, the storage characteristics were evaluated after the shortterm and long-term storage of lotus root at 4 degrees C and 20 degrees C. The hardness of lotus root significantly decreased at both temperatures (4 degrees C and 20 degrees C) during the first 3 days of storage. In contrast, the decrease in hardness delayed at 4 degrees C (beyond 3 days of storage). Further, genes related to hardness at different storage temperatures were identified using the RNA-seq and qRT-PCR. The results of this study provide a reference for lotus root storage and a basis for the molecular breeding of longterm-storable lotus root.
