Selection of High Laccase-Producing Coriolopsis gallica Strain T906: Mutation Breeding, Strain Characterization, and Features of the Extracellular Laccases

( Xiaoli Xu ),( Lei Feng ),( Zhenya Han ),( Sishi Luo ),( Ai`min Wu ),( Jun Xie ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.9

Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro Nnitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30°C and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

Carbon-based electrode fabrication and performance of bioelectrochemical anaerobic digestion for sewage sludge

Qing Feng,Zhengkai An,Rusong Zhao,Xiaoli Wang,Jun Li,송영채 한국마린엔지니어링학회 2019 한국마린엔지니어링학회지 Vol.43 No.10

Bioelectrochemical anaerobic digestion (BEAD) is a technology which exploits the potential of microorganism by supplying a small amount of electric energy in an anaerobic digester. The electrode material is one of the most important parts of a bioelectrochemical system. For the anodes and cathodes of BEAD process, graphite fiber fabric (GFF), a carbon based material with good biocompatibility and non-corrosive property, was selected and its conductivity was modified by depositing carbon nanotube (CNT) via electrophoresis deposition (EPD) method. In the BEAD process, the organic matter in the reactor is oxidized on the surface of anode forming proton, carbon dioxide and electron. The electron produced is transferred to the cathode due to the potential difference between anode and cathode, followed by methane production on the surface of cathode resulting from the combination of carbon dioxide, electron and proton. The biochemical reactions at the electrodes can be controlled by the electrode potentials. When the potential between anode and cathode was maintained at 0.3 V using an external power supply, the BEAD reactor showed remarkable performance after the start-up period of 40 days. The performance of BEAD reactor was stable in terms of pH (7.2~7.5), alkalinity (4,500~5,200mg/L as CaCO3), methane content in biogas (77.3%) and volatile fatty acid levels (VFA < 250mg HAc/L). During steady state, the specific methane production rate and VS reduction were stabilized at 412 mL CH4/L.d and 72.5%, respectively, which were much higher than the conventional anaerobic digestion. The application of bioelectrochemical technology to anaerobic digestion provides a chance to overcome the disadvantages of conventional anaerobic digestion.

Rapid Detection of Campylobacter jejuni Using Fluorescent Microspheres as Label for Immunochromatographic Strip Test

Di Xu,Xiaoli Wu,Bo Li,Peng Li,Xing Ming,Tingtao Chen,Hua Wei,Feng Xu 한국식품과학회 2013 Food Science and Biotechnology Vol.22 No.2

Campylobacter jejuni is a worldwide foodborne pathogen recognized as a leading cause of human gastrointestinal enteritis. A rapid, sensitive, and specific method is required to monitor food and water in cases of contamination by this pathogen. This report presents a novel immunochromatographic test (ICT) using fluorescent microspheres labeled with polyclonal antibodies of C. jejuni as the capture reagent dispensed onto the conjugate pad. Polyclonal antibodies against the outer membrane protein PEB1 of C. jejuni were used as the detective reagent at the test line, whereas the goat anti-rabbit IgG was used on the control line. PEB1 was obtained by gene cloning and expression to prepare its antibody. In this study, a simple and rapid ICT is reported for detecting C. jejuni for the first time with a detection limit of 106 CFU/mL.

Proteomic analyses on the browning of shade-dried Thompson seedless grape

Liu Fengjuan,Huang Wenshu,Feng Zuoshan,Tao Yongxia,Fan Yingying,He Weizhong,Li XiaoLi,Fang Xiaotong,Wang Cheng,Bai Yujia 한국응용생명화학회 2021 Applied Biological Chemistry (Appl Biol Chem) Vol.64 No.3

China is one of the main producers in the worldwide raisin market. Most China’s raisins are produced in Xinjiang where the Thompson seedless grape ( Vitis vinifera L.cv.Thompson seedless) is the main variety of green raisin. However, the browning of Thompson seedless grape during drying has been well-acknowledged as the primary factor affecting the development of the raisin industry. Data independent acquisition (DIA)-based protein profiling was performed on fresh and shade-dried Thompson seedless grapes. As a result, 5431 proteins were identified, among which the amounts of 739 proteins in fresh grape were found to be significantly different with those in dried grape. The functional annotation based on the Blast2GO showed that the ‘organic substance metabolic process’, ‘regulation of molecular function’, ‘enzyme regulator activity’, and ‘isomerase activity’ related proteins became very active during browning. Further analyses revealed that the browning-related proteins, which with significant different amounts in fresh and in dried grapes, are primarily involved in the phenylpropanoid biosynthesis, tyrosine metabolism, phenylalanine metabolism, oxidative phosphorylation metabolism, plutathione metabolism, peroxisome pathway, and fatty acid degradation. And five random differential proteins were verified with parallel reaction monitoring (PRM). The PRM results were in agreement with the DIA data. The main browning-related proteins of Thompson seedless grape were identified in this study. Their properties were tested, and their roles in the browning mechanism were indicated. This will lay base to a better understanding on the enzymatic browning of Thompson seedless grape, and it will also provide guidance for controlling the quality of Thompson seedless grapes in industry.

IRE1α protects against osteoarthritis by regulating progranulin-dependent XBP1 splicing and collagen homeostasis

Liang Li,Zhang Fengmei,Feng Naibo,Kuang Biao,Fan Mengtian,Chen Cheng,Pan Yiming,Zhou Pengfei,Geng Nana,Li Xingyue,Xian Menglin,Deng Lin,Li Xiaoli,Kuang Liang,Luo Fengtao,Tan Qiaoyan,Xie Yangli,Guo Fen 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Osteoarthritis (OA) is a full-joint, multifactorial, degenerative and inflammatory disease that seriously affects the quality of life of patients due to its disabling and pain-causing properties. ER stress has been reported to be closely related to the progression of OA. The inositol-requiring enzyme 1α/X-box-binding protein-1 spliced (IRE1α/XBP1s) pathway, which is highly expressed in the chondrocytes of OA patients, promotes the degradation and refolding of abnormal proteins during ER stress and maintains the stability of the ER environment of chondrocytes, but its function and the underlying mechanisms of how it contributes to the progression of OA remain unclear. This study investigates the role of IRE1α/ERN1 in OA. Specific deficiency of ERN1 in chondrocytes spontaneously resulted in OA-like cartilage destruction and accelerated OA progression in a surgically induced arthritis model. Local delivery of AdERN1 relieved degradation of the cartilage matrix and prevented OA development in an ACLT-mediated model. Mechanistically, progranulin (PGRN), an intracellular chaperone, binds to IRE1α, promoting its phosphorylation and splicing of XBP1u to generate XBP1s. XBP1s protects articular cartilage through TNF-α/ERK1/2 signaling and further maintains collagen homeostasis by regulating type II collagen expression. The chondroprotective effect of IRE1α/ERN1 is dependent on PGRN and XBP1s splicing. ERN1 deficiency accelerated cartilage degeneration in OA by reducing PGRN expression and XBP1s splicing, subsequently decreasing collagen II expression and triggering collagen structural abnormalities and an imbalance in collagen homeostasis. This study provides new insights into OA pathogenesis and the UPR and suggests that IRE1α/ERN1 may serve as a potential target for the treatment of joint degenerative diseases, including OA.

Using tyrosinase as a tri-modality reporter gene to monitor transplanted stem cells in acute myocardial infarction

Mei Liu,Yichun Wang,Mengting Li,Hongyan Feng,Qingyao Liu,Chunxia Qin,Yongxue Zhang,Xiaoli Lan 생화학분자생물학회 2018 Experimental and molecular medicine Vol.50 No.-

The study aimed to investigate the feasibility of noninvasive monitoring of bone marrow mesenchymal stem cells (MSCs) transduced with the tyrosinase reporter gene for acute myocardial infarction (AMI) with photoacoustic imaging (PAI), magnetic resonance imaging (MRI), and positron emission tomography (PET) in vitro and in vivo. MSCs were transduced with a lentivirus carrying a tyrosinase reporter gene. After transduction, the rate of 18F-5-fluoro-N-(2- [diethylamino]ethyl)picolinamide (18F-5-FPN) uptake was measured. PAI and MRI of stable cell lines expressing tyrosinase (TYR-MSCs) were performed in vitro. An AMI model was induced and verified. TYR-MSCs and MSCs were injected into the margins of the infarcted areas, and PAI, MRI, and PET images were acquired 1, 7, 14, 21, and 28 days after cell injection. Sham-operated models without injection were used as the control group. TYR-MSCs showed noticeably higher uptake of 18F-5-FPN and stronger signals in T1-weighted MRI and PAI than non-transduced MSCs. In vivo studies revealed prominent signals in the injected area of the infarcted myocardium on PAI/MRI/PET images, whereas no signal could be seen in rats injected with non-transduced MSCs or sham-operated rats. The uptake values of 18F-5-FPN in vivo showed a slight decrease over 28 days, whereas MRI and PAI signal intensity decreased dramatically. MSCs stably transduced with the tyrosinase reporter gene could be monitored in vivo in myocardial infarction models by PET, MRI, and PAI, providing a feasible and reliable method for checking the viability, location, and dwell time of transplanted stem cells.

Anatomical Observations of Adventitious Bud Regeneration from Leaf Explants of Ziziphus jujube Mill. ‘Huizao’

Chunhua Ma,Xia Ye,Yanhui Chen,Jiancan Feng,Xiaoli Shang,Jidong Li,Yingxia Wu,Jianbin Hu 한국원예학회 2012 Horticulture, Environment, and Biotechnology Vol.53 No.4

The histological process of adventitious shoot regeneration from the leaf explants of Zizyphus jujuba ‘Huizao’was reported in this study. This is the first report on the detailed histological process of direct shoot regeneration from leaf explants in Z. jujube. Shoot regeneration was obtained from woody plant medium (WPM) supplemented with 2.27 μM thidiazuron (TDZ), 1.07 μM α-naphthalene-acetic acid (NAA) and 2.94 μM silver nitrate (AgNO3) for 10days in the dark followed by 3 weeks at a 14 hours photoperiod. The adventitious buds mostly formed from leaf veins and petioles, and the further histological studies revealed that there were multiple vascular bundles around leaf veins and the adaxial side of explants, and the adventitious buds directly originated from the parenchymatous cells around the vascular bundles without the intervening callus phase. After 3 days of culture, the parenchymatous cells started dividing and meristemoids formed thereafter. The meristematic cells continued division and subsequently gave rise to bud primordia. Well-developed shoot buds through direct organogenesis was achieved after 20 days of culture.

Interference of periostin attenuates pathological changes, proinflammatory markers and renal fibrosis in diabetic kidney injury

Duan Xiaoting,Chen Cheng,Liu Xiaoli,Wang Taoxia,Feng Shuning,Li Jianwei,Li Guiying 한국유전학회 2023 Genes & Genomics Vol.45 No.11

Background Diabetic nephropathy (DN) is a prevalent complication of diabetes, in which inflammation and fibrosis are the significant pathogenesis. Periostin is a matricellular protein that functions on stabilizing the extracellular matrix by binding to integrins during development. This study aimed to explored the role of periostin in DN. Methods The animal and cell models of DN were constructed in streptozocin (STZ)-induced mice and high glucose-challenged human mesangial cells (HMCs). The role of periostin in pathological changes, inflammation and fibrosis in DN was investigated through biochemical detection, HE and Masson staining and scores, western blot, enzyme‑linked immunosorbent assay (ELISA) and real-time quantitative PCR (RT-qPCR) assays. Results Knockdown of periostin counteracted the STZ-induced the ratio of kidney weight and body weight, and the concentrations of urine albumin excretion (UAE), serum creatinine (Scr), urine albumin/creatinine ratio (UACR) and blood urea nitrogen (BUN) in mice. Moreover, silencing of periostin alleviated the pathological manifestations and reduced the concentrations of IL-6, TNF-α and IL-1β in mice kidney tissues and sera. Also, downregulation of periostin decreased the relative protein expression of fibronectin, collagen IV and α-SMA in kidney tissues. Meanwhile, interference of periostin attenuated the levels of pro-inflammation factors and the expressions of fibrosis markers in HG-induced HMCs. Conclusion Interference of periostin resisted DN via attenuating the pro-inflammatory cytokines release and renal fibrosis in diabetic kidney injury. Our study establishes a basis for its further study and underlying application in clinical practice in diagnosing and treating diabetic kidney injury or other relevant diseases.

Identification of MicroRNAs as potential biomarkers for detecting ischemic stroke

Li Kexin,Shen Li,Zheng Pingping,Wang Yanjun,Wang Lijuan,Meng Xiaoli,Lv Yaogai,Xue Zhiqiang,Guo Xin,Zhang Anning,Pan Pan,Bi Chunli,Chen Yang,Feng Tianyu,Li Bo,Jin Lina,Yao Yan 한국유전학회 2022 Genes & Genomics Vol.44 No.1

Background: Increasing epidemic of ischemic stroke (IS) makes it urgent to understand the pathogenesis and regulatory mechanism, previous studies have described microRNAs (miRNAs) is part of the brain's response to ischemia. Objective: The aim of this study was to screen potential biomarkers for the prediction and novel treatment of IS. Methods: Differentially expressed miRNAs were screened from three newly diagnosed IS patients and three controls by RNA sequencing technology. Furthermore, target prediction databases were then used to analysis the target genes of different expressed miRNAs, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database were used to identify the functions and the main biochemical and signal pathways of differentially expressed target genes. Results: Our results revealed that 27 miRNAs were differentially expressed in IS, among which, hsa-miR-659-5p was the most highly increased and was first found to be associated with IS. In addition, KEGG pathway analyses showed that differentially expressed miRNAs were mainly significantly enriched in lysosome pathway, cytokine-cytokine receptor interaction pathway, spliceosome pathway, base excision repair pathway. Conclusions: miRNAs were involved in IS pathogenesis, and hsa-miR-659-5p, hsa-miR-151a-3p and hsa-miR-29c-5p as the three highest |log2FoldChange| regulation in this study, which may be the biomarkers of IS and need further study.

Anterior Gradient 3 Promotes Breast Cancer Development and Chemotherapy Response

Qiao Xu,Ying Shao,Jinman Zhang,Huikun Zhang,Yawen Zhao,Xiaoli Liu,Zhifang Guo,Wei Chong,Feng Gu,Yongjie Ma 대한암학회 2020 Cancer Research and Treatment Vol.52 No.1

Purpose Anterior gradient 3 (AGR3) belongs to human anterior gradient (AGR) family. The function of AGR3 on cancer remains unknown. This research aimed to investigate if AGR3 had prognostic values in invasive ductal carcinoma (IDC) of breast cancer and could promote tumor progression. Materials and Methods AGR3 expression was detected in breast benign lesions, ductal carcinoma in situ and IDC by immunohistochemistry analysis. AGR3’s correlations with clinicopathological features and prognosis of IDC patients were analyzed. By cell function experiments, collagen gel droplet-embedded culture drug sensitivity test and cytotoxic analysis, AGR3’s impacts on proliferation, invasion ability, and chemotherapeutic drug sensitivity of breast cancer cells were also detected. Results AGR3 was up-regulated in luminal subtype of histological grade I-II of IDC patients and positively correlated with high risks of recurrence and distant metastasis. AGR3 high expression could lead to bone or liver metastasis and predict poor prognosis of luminal B. In cell lines, AGR3 could promote proliferation and invasion ability of breast cancer cells which were consistent with clinical analysis. Besides, AGR3 could indicate poor prognosis of breast cancer patients treated with taxane but a favorable prognosis with 5-fluoropyrimidines. And breast cancer cells with AGR3 high expression were resistant to taxane but sensitive to 5-fluoropyrimidines. Conclusion AGR3 might be a potential prognostic indicator in luminal B subtype of IDC patients of histological grade I-II. And patients with AGR3 high expression should be treated with chemotherapy regimens consisting of 5-fluoropyrimidines but no taxane.