Comparison of structural features and antioxidant activity of polysaccharides from natural and cultured Cordyceps sinensis

Junqiao Wang,Shaoping Nie,Lijiao Kan,Haihong Chen,Steve W. Cui,Aled O. Phillips,Glyn O. Phillips,Mingyong Xie 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.1

Four polysaccharides (named as P1, P2, and P3 from three natural Cordyceps sinensis and P4 from cultured C. sinensis) were obtained by hot-water extraction and ethanol precipitation and their structural characteristics as well as antioxidant potentials were compared. Results revealed that the backbone of P1, P2, and P3 comprised α-1,4-glucose, with a branching point mainly at position 6 and terminating at glucose. On the other hand, the structure of P4 was highly complex, mainly comprising glucose, galactose, and mannose, with 1,4-glucose and 1,4-galactose as the main chain. For in vitro antioxidant assays, all the four polysaccharides showed similar scavenging capacity against DPPH and hydroxyl radicals, whereas P1 had a relatively low ferric reducing ability, possibly related to a combination of factors such as the phenolic compounds and amino acids that conjugated in polysaccharides.

An Effective Method for Deproteinization of Bioactive Polysaccharides Extracted from Lingzhi (Ganoderma atrum)

Yi Chen,Mingyong Xie,Wenjuan Li,Hui Zhang,Shaoping Nie,Yuanxing Wang,Chang Li 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.1

Deproteinization procedure is a fundamental step for analyzing polysaccharide from natural plants. In this study, in the course of refining bioactive polysaccharides from lingzhi (Ganoderma atrum), an effective deproteinization method using lead acetate solution was established by comparing with other available methods. The percentages of deproteinization, polysaccharide loss, and its antioxidant activities loss were used as the index to evaluate and optimize the precipitation experimental conditions. The results showed that the modified method, precipitation with the addition of 0.4-0.52%(w/v) lead acetate, was superior to the others, as evidenced by the highest deproteinization efficiency (88%), as well as the lowest polysaccharide loss (17%). And notably its antioxidant activity also remained good (loss 15%). It provides a simple prefractionation step for the analysis of polysaccharide from natural plants. Polysaccharide isolated by this method is in the native state. Our method may offer a rapid method for removing protein from plant polysaccharides in large scale.

An efficient protocol for Agrobacterium-mediated transformation of the biofuel plant Jatropha curcas by optimizing kanamycin concentration and duration of delayed selection

Qiantang Fu,Chaoqiong Li,Mingyong Tang,Yan-Bin Tao,Bang-Zhen Pan,Lu Zhang,Longjian Niu,Huiying He,Xiulan Wang,Zeng-Fu Xu 한국식물생명공학회 2015 Plant biotechnology reports Vol.9 No.6

Jatropha curcas is considered a potential biodiesel feedstock crop. Currently, the value of J. curcas is limited because its seed yield is generally low. Transgenic modification is a promising approach to improve the seed yield of J. curcas. Although Agrobacterium-mediated genetic transformation of J. curcas has been pursued for several years, the transformation efficiency remains unsatisfying. Therefore, a highly efficient and simple Agrobacterium-mediated genetic transformation method for J. curcas should be developed. We examined and optimized several key factors that affect genetic transformation of J. curcas in this study. The results showed that the EHA105 strain was superior to the other three Agrobacterium tumefaciens strains for infecting J. curcas cotyledons, and the supplementation of 100 mM acetosyringone slightly increased the transient transformation frequency. Use of the appropriate inoculation method, optimal kanamycin concentration and appropriate duration of delayed selection also improved the efficiency of stable genetic transformation of J. curcas. The percentage of b-glucuronidase positive J. curcas shoots reached as high as 56.0 %, and 1.70 transformants per explant were obtained with this protocol. Furthermore, we optimized the root-inducing medium to achieve a rooting rate of 84.9 %. Stable integration of the T-DNA into the genomes of putative transgenic lines was confirmed by PCR and Southern blot analysis. Using this improved protocol, a large number of transgenic J. curcas plantlets can be routinely obtained within approximately 4 months. The detailed information provided here for each step of J. curcas transformation should enable successful implementation of this transgenic technology in other laboratories.

Pim-1 Protects Retinal Ganglion Cells by Enhancing Their Regenerative Ability Following Optic Nerve Crush

Shoumei Zhang,Li Shuai,Dong Wang,Tingting Huang,Shengsheng Yang,Mingyong Miao,Fang Liu,Jiajun Xu 한국뇌신경과학회 2020 Experimental Neurobiology Vol.29 No.3

Provirus integration site Moloney murine leukemia virus (Pim-1) is a proto-oncogene reported to be associated with cell proliferation, differentiation and survival. This study was to explore the neuroprotective role of Pim-1 in a rat model subjected to optic nerve crush (ONC), and discuss its related molecules in improving the intrinsic regeneration ability of retinal ganglion cells (RGCs). Immunofluorescence staining showed that AAV2- Pim-1 infected 71% RGCs and some amacrine cells in the retina. Real-time PCR and Western blotting showed that retina infection with AAV2- Pim-1 up-regulated the Pim-1 mRNA and protein expressions compared with AAV2-GFP group. Hematoxylin-Eosin (HE) staining, γ-synuclein immunohistochemistry, Cholera toxin B (CTB) tracing and TUNEL showed that RGCs transduction with AAV2-Pim-1 prior to ONC promoted the survival of damaged RGCs and decreased cell apoptosis. RITC anterograde labeling showed that Pim-1 overexpression increased axon regeneration and promoted the recovery of visual function by pupillary light reflex and flash visual evoked potential. Western blotting showed that Pim- 1 overexpression up-regulated the expression of Stat3, p-Stat3, Akt1, p-Akt1, Akt2 and p-Akt2, as well as βIII-tubulin, GAP-43 and 4E-BP1, and downregulated the expression of SOCS1 and SOCS3, Cleaved caspase 3, Bad and Bax. These results demonstrate that Pim-1 exerted a neuroprotective effect by promoting nerve regeneration and functional recovery of RGCs. In addition, it enhanced the intrinsic regeneration capacity of RGCs after ONC by activating Stat3, Akt1 and Akt2 pathways, and inhibiting the mitochondrial apoptosis pathways. These findings suggest that Pim-1 may prove to be a potential therapeutic target for the clinical treatment of optic nerve injury

The Differences between Luminal Microbiota and Mucosal Microbiota in Mice

Minna Wu,Puze Li,Jianmin Li,Yunying An,Mingyong Wang,Genshen Zhong 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.2

The differences between luminal microbiota (LM) and mucosal microbiota (MAM) were little known, especially in duodenum. In this study, LM and MAM in colon and duodenum of mice were investigated through 16S rRNA high-throughput sequencing. The lowest bacterial diversity and evenness were observed in duodenal LM (D_LM), followed by duodenal MAM (D_MAM). Meanwhile, the bacterial diversity and evenness were obviously increased in D_MAM than these in D_LM, while no significant difference was observed between colonic MAM (C_MAM) and colonic LM (C_LM). PCoA analysis also showed that bacterial communities of LM and MAM in duodenum were completely separated, while these in colon overlapped partly. The ratio of Firmicutes to Bacteroidetes (F/B) in D_MAM was significantly higher than that in D_LM. Lactobacillus was largely enriched and was the characteristic bacteria in D_LM. The characteristic bacteria in D_MAM were Turicibacter, Parasutterella, Marvinbryantia and Bifidobacterium, while in C_LM they were Ruminiclostridium_6, Ruminiclostridium_9, Ruminococcaceae_UCG_007 and Lachnospiraceae_UCG_010, and in C_MAM they were Lachnospiraceae_NK4A136, Mucispirillum, Alistipes, Ruminiclostridium and Odoribacter. The networks showed that more interactions existed in colonic microbiota (24 nodes and 74 edges) than in duodenal microbiota (17 nodes and 29 edges). The 16S rDNA function prediction results indicated that bigger differences of function exist between LM and MAM in duodenum than these in colon. In conclusion, microbiota from intestinal luminal content and mucosa were different both in colon and in duodenum, and bacteria in colon interacted with each other much more closely than those in duodenum.

Interplay between the Gut Microbiome and Metabolism in Ulcerative Colitis Mice Treated with the Dietary Ingredient Phloretin

( Jie Ren ),( Puze Li ),( Dong Yan ),( Min Li ),( Jinsong Qi ),( Mingyong Wang ),( Genshen Zhong ),( Minna Wu ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.10

A growing number of healthy dietary ingredients in fruits and vegetables have been shown to exhibit diverse biological activities. Phloretin, a dihydrochalcone flavonoid that is abundant in apples and pears, has anti-inflammatory effects on ulcerative colitis (UC) mice. The gut microbiota and metabolism are closely related to each other due to the existence of the food-gut axis in the human colon. To investigate the interplay of faecal metabolites and the microbiota in UC mice after phloretin treatment, phloretin (60 mg/kg) was administered by gavage to ameliorate dextran sulfate sodium (DSS)-induced UC in mice. Gut microbes and faecal metabolite profiles were detected by high-throughput sequencing and liquid chromatography mass spectrometry (LC-MS) analysis, respectively. The correlations between gut microbes and their metabolites were evaluated by Spearman correlation coefficients. The results indicated that phloretin reshaped the disturbed faecal metabolite profile in UC mice and improved the metabolic pathways by balancing the composition of faecal metabolites such as norepinephrine, mesalazine, tyrosine, 5-acetyl-2,4- dimethyloxazole, and 6-acetyl-2,3-dihydro-2-(hydroxymethyl)-4(1H)-pyridinone. Correlation analysis identified the relations between the gut microbes and their metabolites. Proteus was negatively related to many faecal metabolites, such as norepinephrine, L-tyrosine, laccarin, dopamine glucuronide, and 5-acetyl-2,4-dimethyloxazole. The abundance of unidentified Bacteriodales_S24-7_group was positively related to ecgonine, 15-KETE and 6-acetyl-2,3-dihydro-2- (hydroxymethyl)-4(1H)-pyridinone. The abundance of Christensenellaceae_R-7_group was negatively related to the levels of 15-KETE and netilmicin. Stenotrophomonas and 15-KETE were negatively related, while Intestinimonas and alanyl-serine were positively related. In conclusion, phloretin treatment had positive impacts on faecal metabolites in UC mice, and the changes in faecal metabolites were closely related to the gut microbiota.