Efficient Expression, Purification, and Characterization of a Novel FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus in Pichia pastoris

( Yu Feng Yang ),( Lei Huang ),( Ju Fang Wang ),( Xiao Ning Wang ),( Zhi Nan Xu ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.11

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and 55°C. In addition, it displayed very high thermal stability, with a half-life of 82 min at 60°C. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

A Comparison of Developmental Trajectories of Prepulse Inhibition between Male and Female Rats

Yang Yang,Yun'ai Su,Chunmei Guo,Yu Feng,Jitao Li,Tianmei Si 대한정신약물학회 2010 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.8 No.3

Objective: Various studies have demonstrated that childhood and adolescence are critical periods of brain development characterized by both dramatic changes and sexual dimorphism in anatomy, physiology and cognition. Prepulse inhibition (PPI) is an operational model of sensorimotor gating process, but data concerning the interactions of sex and normal ontogeny of PPI is of a particular paucity. Methods: We led a longitudinal study to compare the PPI between male and female Sprague-Dawley rats during postnatal day 23 to 43. Results: We found that the PPI of both male and female rats increased during this period. We also found that the PPI of male and female rats had different developmental trajectories when the prepulse intensity was 20 dB [A] above background, a result similar with a previous study done in humans. Conclusion: The finding confirms the face validity of PPI as a model of information processing across species and supports its utilization for investigation of potential aetiological factors implicated in mental disorders.

Mitochondrial citrate accumulation drives alveolar epithelial cell necroptosis in lipopolysaccharide-induced acute lung injury

Yang Hui-Hui,Jiang Hui-Ling,Tao Jia-Hao,Zhang Chen-Yu,Xiong Jian-Bing,Yang Jin-Tong,Liu Yu-Biao,Zhong Wen-Jing,Guan Xin-Xin,Duan Jia-Xi,Zhang Yan-Feng,Liu Shao-Kun,Jiang Jian-Xin,Zhou Yong,Guan Cha-Xi 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Necroptosis is the major cause of death in alveolar epithelial cells (AECs) during acute lung injury (ALI). Here, we report a previously unrecognized mechanism for necroptosis. We found an accumulation of mitochondrial citrate (citratemt) in lipopolysaccharide (LPS)-treated AECs because of the downregulation of Idh3α and citrate carrier (CIC, also known as Slc25a1). shRNA- or inhibitor–mediated inhibition of Idh3α and Slc25a1 induced citratemt accumulation and necroptosis in vitro. Mice with AEC-specific Idh3α and Slc25a1 deficiency exhibited exacerbated lung injury and AEC necroptosis. Interestingly, the overexpression of Idh3α and Slc25a1 decreased citratemt levels and rescued AECs from necroptosis. Mechanistically, citratemt accumulation induced mitochondrial fission and excessive mitophagy in AECs. Furthermore, citratemt directly interacted with FUN14 domain-containing protein 1 (FUNDC1) and promoted the interaction of FUNDC1 with dynamin-related protein 1 (DRP1), leading to excessive mitophagy-mediated necroptosis and thereby initiating and promoting ALI. Importantly, necroptosis induced by citratemt accumulation was inhibited in FUNDC1-knockout AECs. We show that citratemt accumulation is a novel target for protection against ALI involving necroptosis.

Prognostic Role of Nucleophosmin in Colorectal Carcinomas

Yang, Yu-Feng,Zhang, Xi-Ying,Yang, Mei,He, Ze-Hua,Peng, Ning-Fu,Xie, Shu-Rui,Xie, Yan-Fang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.5

Aim: Recent research suggests that nucleophosmin (NPM) may be a prognostic marker in colorectal carcinomas (CRC). We here tested its use to predict the survival of CRC patients. Methods: We investigated NPM expression by immunohistochemistry in histologically normal to malignant colorectal tissues and evaluated its association with clinicopathological variables. Overall and disease-free survival after tumor removal were calculated by the Kaplan-Meier method, and differences in survival curves were analyzed by the log-rank test. The Cox proportional hazards model was used for multivariate analysis of prognostic factors. Results: NPM expression was found significantly upregulated in CRC compared to adjacent colorectal tissue, villous adenoma, tubular adenoma and normal colorectal mucosa (p<0.05 for all). NPM expression was statistically linked to cancer embolus, lymph node metastasis, differentiation grade, and recurrence of CRC. Overall and disease-free survival of NPM-negative CRC patients tended to be better than those for patients with NPM-positive lesions (log-rank statistic, p<0.05 for all). Multivariate analysis indicated NPM expression as an independent prognostic indicator for CRC patients (p<0.05 ). Conclusion: Our results suggest that NPM expression can predict the survival of CRC patients. Prognosis of CRC is determined by not only many known prognostic factors but also by NPM expression.

DEVELOPMENT OF CULTIVATAED SEAWEED Gracilaria AND ITS CONTRIBUTION TO ENVIRONMENTAL IMPROVEMENTS IN CHINESE COASTAL WATERS

Yu Feng Yang,XiuGeng Fei,Ik Kyo Chung 한국수산과학회 양식분과 2015 한국수산과학회 양식분과 학술대회 Vol.2015 No.5

New Permeable Structure for Controlling Debris Flows in the Wenjiagou Gully

Yu-feng Liang,Chuan Liang,Hongwei Zhou,Feng-rong Yang,Miao Huo,Jia-wen Zhou 대한토목학회 2018 KSCE JOURNAL OF CIVIL ENGINEERING Vol.22 No.11

Aimed to control debris flows more effectively, we attempt to develop a new structure called a “sloping roof grill dam”, which consists of guidance walls, separation grills and lateral deposit areas. A series of flume tests were performed to verify the function of this new structure, focusing on the trapping efficiency (β ), deceleration efficiency (μ) and coarse-fine sediment separation extents (η) under different conditions of structure and debris flows. Experimental results show that both the trapping and deceleration efficiency increase with a decrease in permeability, whereas the separation extent shown the opposite effect. Multiple regression analysis indicates that β and η are linearly dependent on the permeability, deceleration efficiency and magnitudes of debris flow. In addition, an increase in beam gap can lead to an increase in the impact area downstream for the debris flow after the control of the structure. Finally, a two-level installation of the structure is tested to determine whether there is an enhancement of the debris control, which indicated that although the trapping effect of the two-level structure is considerably greater than that of a single-level structure (2.11 times at maximum), the separation extent of the two-level structure is not distinct and may be even lower.

Prevalence and Age, Gender and Geographical Area Distribution of Esophageal Squamous Cell Carcinomas in North China from 1985 to 2006

Feng, Xiao-Shan,Yang, Yan-Tong,Gao, She-Gan,Ru, Yi,Wang, Gong-Ping,Zhou, Bo,Wang, Yu-Feng,Zhang, Peng-Fei,Li, Pu-Yu,Liu, Yong-Xuan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.5

Objective: To establish the prevalence and distribution profile of esophageal squamous cell carcinomas (ESCCs) over a 22-yr period in North China. Methods: Using endoscopy for primary diagnosis and histological analysis for the further confirmation, a total of 74,854 ESCC patients aged 20-89 between January 1985 and December 2006 were investigated to analyze the epidemiological profile including prevalence rates, distribution of age-of-onset, gender and geographical area of ESCC in Luoyang, the highest incidence area of North China. Results: A total of 4092 cases of ESCC were finally diagnosed among 74,854 patients who had their first endoscopies. The prevalence among males was higher than that among females (p<0.01), resulting in an overall male:female OR of 1.2 (95%CI, 1.2-1.3). The prevalence in rural areas was higher than in urban areas (p<0.01), resulting in an overall rural:urban OR of 2.6 (95%CI, 2.4-2.9). The rural:urban ORs and the 95% CI increased continuously from 2.6, 2.3-3.0 to 2.7, 2.2-3.3, respectively, for 4 consecutive periods during the 22-yr study period. Moreover, the median age of onset among females was higher than that among males (p<0.01). For both sexes and in both areas, the prevalence rates declined and the median age of onset rose for 4 consecutive periods in the 22-yrs time frame (p<0.01). Conculsions: These data reveal the epidemiological profile of ESCC in the area of North China, and suggest that urban areas and rural people account for a growing proportion of the ESCC patients although the prevalence of ESCC significantly declined and the median age-of-onset postponed over the 22-yrs period. Moreover, the prevalence status of ESCC in rural areas also underlines the need for public health initiatives aimed at reducing risk factors of this fatal disease.

Isoindolin-1-ones from the stems of Nicotiana tabacum and their antiviral activities

Guang-Yu Yang,Jia-Meng Dai,Zhen-Jie Li,Jin Wang,Feng-Xian Yang,Xin Liu,Jing Li,Qian Gao,Xue-Mei Li,Yin-Ke Li,Wei-Guang Wang,Min Zhou,Qiu-Fen Hu 대한약학회 2022 Archives of Pharmacal Research Vol.45 No.8

In previous studies, several isoindolin-1-oneanalogs that exhibited signifi cant anti-tobacco mosaic virus(anti-TMV) activities were isolated from Nicotiana tabacum . Since gene-editing mutants provide a new sample for thediscovery of active metabolites, we focused on the stems ofYN-18–23 (a mutant N. tabacum for gene editing with thealkaloid metabolic pathway cultivated by Yunnan TobaccoCompany), which led to the isolation of four new ( 1–4 )and four known ( 5–8 ) isoindolin-1-ones. To the best of ourknowledge, nicindole C ( 3 ) is the fi rst subclass of isoindolin-1-one bearing a pentacyclic ketone, while nicindole D ( 4 )is the fi rst example of isoindolin-1-one bearing a methylpyridin-2-(1 H )-one moiety. Compounds 1–4 were testedfor their anti-TMV activities, and the results revealed thatcompounds 1 , 3 , and 4 exhibited high anti-TMV activities atconcentrations of 20 μM with inhibition rates of 48.6, 42.8,and 71.5%, respectively. These rates are higher than the inhibitionrate of the positive control (33.2%). The mechanisticstudy of compound 4 , which had the highest anti-TMV activityrevealed that increased potentiation of defense-related enzyme activities and downregulation of expression of theNtHsp70 protein may induce resistance in tobacco againstthe viral pathogen TMV. Molecular docking studies alsorevealed that the isoindolin-1-one substructure is fundamentalfor anti-TMV activity. The methyl-pyridin-2-(1 H )-onemoiety in compound 4 and the 2-oxopropyl groups in compounds1 and 3 at the N -2 position may increase inhibitoryactivities. This study of the structure–activity relationshipis helpful for fi nding new anti-TMV activity inhibitors. Tostudy whether the isoindolin-1-ones have broader antiviralactivities, compounds 1–4 were also tested for their antirotavirusactivities. Compound 4 exhibited high anti-rotavirusactivity with a therapeutic index (TI) value of 20.7. This TI value is close to that of the positive control (20.2).

Intense red photoluminescence and mechanoluminescence from Mn2+-activated SrZnSO with a layered structure

Zhou, Yu,Yang, Yun-Ling,Fan, Yu-Ting,Yang, Woochul,Zhang, Wei-Bin,Hu, Jian-Feng,Zhang, Zhi-Jun,Zhao, Jing-Tai The Royal Society of Chemistry 2019 Journal of Materials Chemistry C Vol.7 No.26

<P>A series of novel red emitting Mn<SUP>2+</SUP>-activated SrZnSO phosphors were successfully synthesized by solid-state reaction at high temperature. The photoluminescence (PL) and mechanoluminescence (ML) properties of these Mn<SUP>2+</SUP>-activated SrZnSO phosphors with different Mn<SUP>2+</SUP> concentrations were investigated. With increasing the concentration of Mn<SUP>2+</SUP> from <I>x</I> = 0 to 0.04, the unit cell volume increased from 153.82 to 154.19 Å<SUP>3</SUP> while the optical band gap decreased from 3.74 to 3.43 eV. The site occupation of Mn<SUP>2+</SUP> in the host lattice was demonstrated by Rietveld refinement, the electron paramagnetic resonance (EPR) spectrum, and the spectroscopic properties. A broad band emission peak at 603 nm of SrZn1−xMnxSO (0.001 ≤ <I>x</I> ≤ 0.04) with an excitation wavelength of 318 nm was attributed to electronic transitions of Mn<SUP>2+</SUP> from the <SUP>4</SUP>T1(<SUP>4</SUP>G) level to the <SUP>6</SUP>A1(<SUP>6</SUP>S) level. The lifetime of SrZn1−xMnxSO (0.001 ≤ <I>x</I> ≤ 0.04) decreased monotonously from 2.97 to 0.82 ms with increasing Mn<SUP>2+</SUP> concentration. In particular, intense emission of red light from SrZn1−xMnxSO (0.001 ≤ <I>x</I> ≤ 0.04) under compressive load could be observed even with the naked eye, indicating that SrZn1−xMnxSO could be used for stress sensors or stress imaging. There was a linear correlation between the ML intensity and external load in SrZn1−xMnxSO, and the ML intensity could be recovered under UV light irradiation. Considering its advantages of non-destruction, reproducibility, and high ML intensity, SrZn1−xMnxSO might be useful for non-destructive detection of stress.</P>

Two Compound Heterozygous Were Identified in SLC26A4 Gene in Two Chinese Families With Enlarged Vestibular Aqueduct

Yongbo Yu,Yang Yang,Jie Lu,Yaqiong Jin,Yeran Yang,Enyu Hong,Jin Shi,Feng Chen,Shujing Han,Ping Chu,Yongli Guo,Xin Ni 대한이비인후과학회 2019 Clinical and Experimental Otorhinolaryngology Vol.12 No.1

Objectives. To investigate the genetic causes of hearing loss with enlarged vestibular aqueduct (EVA) in two children from unrelated two Chinese families. Methods. Sanger sequencing of all coding exons in SLC26A4 (encoding Pendrin protein) was performed on the two patients, their sibling and parents respectively. To predict and visualize the potential functional outcome of the novel variant, model building, structure analysis, and in silico analysis were further conducted. Results. The results showed that the proband from family I harbored a compound heterozygote of SLC26A4 c.1174A>T (p.N392Y) mutation and c.1181delTCT (p.F394del) variant in exon 10, potentially altering Pendrin protein structure. In family II, the proband was identified in compound heterozygosity with a known mutation of c.919-2A>G in the splice site of intron 7 and a novel mutation of c.1023insC in exon 9, which results in a frameshift and translational termination, consequently leading to truncated Pendrin protein. Sequence homology analysis indicated that all the mutations localized at high conservation sites, which emphasized the significance of these mutations on Pendrin spatial organization and function. Conclusion. In summary, this study revealed two compound heterozygous mutations (c.1174A>T/c.1181delTCT; c.919-2A>G/c.1023insC) in Pendrin protein, which might account for the deafness of the two probands clinically diagnosed with EVA. Thus this study contributes to improve understanding of the causes of hearing loss associated with EVA and develop a more scientific screening strategy for deafness.