Optimized Transformation of Streptomyces sp. ATCC 39366 Producing Leptomycin by Electroporation

Yong-Qiang Fan,Hong-Jian Liu,Li Yan,Yu-Shi Luan,Hai-Meng Zhou,Jun-Mo Yang,Shang-Jun Yin,Yu-Long Wang 한국미생물학회 2013 The journal of microbiology Vol.51 No.3

Streptomyces sp. ATCC 39366 produces leptomycin derivatives. Leptomycin B, a potent and specific inhibitor against the export of nuclear proteins, is the main product; however, the introduction of DNA into this strain is almost impossible, which has impeded its further use. We developed a Streptomyces sp. ATCC 39366 transformation protocol to introduce foreign DNA via electroporation. Various conditions were examined, including treatments of the cell wall with weakening agents, electroporation parameters, and DNA content. We found that only plasmid DNA isolated from a dam- ET12567 strain resulted in successful transformation. The mycelium growing in a yeast-peptone-dextrose medium supplemented with 1% glycine at 28°C on a rotary shaker (220 rpm) was more dispersed than those without supplementation and prone to electroporation. The maximum transformation efficiency of 8×102 CFU/μg plasmid DNA was obtained at a field strength of 13 kV/cm with a time constant of 13 ms (25-μF capacitor; parallel resistance, 600 Ω) using 1-mm electrocuvettes. The results of the transformations of two other Streptomyces species indicated that the optimized conditions established in this study might only be applicable to Streptomyces sp. ATCC 39366. However, this is the first report of successful transformation of Streptomyces sp. ATCC 39366, and will facilitate the construction of a gene knockout mutant in Streptomyces sp. ATCC 39366 to produce series of new leptomycin derivatives.

Inactivation of recombinant human brain-type creatine kinase during denaturation by guanidine hydrochloride in a macromolecular crowding system.

Fan, Yong-Qiang,Liu, Hong-Jian,Li, Chang,Luan, Yu-Shi,Yang, Jun-Mo,Wang, Yu-Long Humana Press 2013 Applied biochemistry and biotechnology Vol.169 No.1

<P>In this study, we quantitatively examined the effects of the macromolecular crowding agents, polyethylene glycol 2000 (PEG 2000) and dextran 70, on guanidine hydrochloride (GdnHCl)-induced denaturation of recombinant human brain-type creatine kinase (rHBCK). Our results showed that both PEG 2000 and dextran 70 had a protective effect on the inactivation of rHBCK induced by 0.5 M GdnHCl at 25 C. The presence of 200 g/L PEG 2000 resulted in the retention of 35.33 % of rHBCK activity after 4 h of inactivation, while no rHBCK activity was observed after denaturation in the absence of macromolecular crowding agents. The presence of PEG 2000 and dextran 70 at a concentration of 100 g/L could decelerate the k (2) value of the slow track to 21 and 33 %, respectively, in comparison to values obtained in the absence of crowding agents. Interestingly, inactivation of rHBCK in the presence of 200 g/L PEG 2000 followed first-order monophasic kinetics, with an apparent rate constant of 8??10(-5)?s(-1). The intrinsic fluorescence results showed that PEG 2000 was better than dextran 70 at stabilizing rHBCK conformation. In addition, the results of the phase diagram indicate that more intermediates may be captured when rHBCK is denatured in a macromolecular crowding system. Mixed crowding agents did not produce better results than single crowding agents, but the protective effects of PEG 2000 on the inactivation and unfolding of rHBCK tended to increase as the ratio of PEG 2000 increased in the mixed crowding agent solution. Though it is not clear which crowding agents more accurately simulated the intracellular environment, this study could lead to a better understanding of protein unfolding in the intracellular environment.</P>

Prebiotic Potential of Xylooligosaccharides Derived from Corn Cobs and Their In Vitro Antioxidant Activity When Combined with Lactobacillus

( Xiuhua Yu ),( Jianyuan Yin ),( Lin Li ),( Chang Luan ),( Jian Zhang ),( Chunfang Zhao ),( Shengyu Li ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.7

In the present work, the in vitro prebiotic activity of xylooligosaccharides (XOS) derived from corn cobs combined with Lactobacillus plantarum, a probiotic microorganism, was determined.These probiotics exhibited different growth characteristics depending on strain specificity. L. plantarum S2 cells were denser and their growth rates were higher when cultured on XOS. Acetate was found to be the major short-chain fatty acid produced as the end-product of fermentation, and its amount varied from 1.50 to 1.78 mg/ml. The antimicrobial activity of XOS combined with L. plantarum S2 was determined against gastrointestinal pathogens. The results showed that XOS proved to be an effective substrate, enhancing antimicrobial activity for L. plantarum S2. In vivo evaluation of the influence of XOS and L. plantarum S2, used both alone and together, on the intestinal microbiota in a mouse model showed that XOS combined with L. plantarum S2 could increase the viable lactobacilli and bifidobacteria in mice feces and decrease the viable Enterococcus, Enterobacter, and Clostridia spp. Furthermore, in the in vitro antioxidant assay, XOS combined with L. plantarum S2 possessed significant 2,2-diphenyl-1- picrylhydrazyl, 2,2’-azino-bis, and superoxide anion radical-scavenging activities, and the combinations showed better antioxidant activity than either XOS or L. plantarum S2 alone.

Positive Association of Human SHC3 Gene with Schizophrenia in a Northeast Chinese Han Population

Ye Lv,Yang Sun,Guan-Yu Wang,Jian Yin,Cheng-Jie Li,Yi-Yang Luo,Zhi-Lin Luan 대한신경정신의학회 2020 PSYCHIATRY INVESTIGATION Vol.17 No.9

Objective Schizophrenia is one of the most devastating neuropsychiatric disorders. Genetic epidemiological studies have confirmed that schizophrenia is a genetic disease. Genes promoting neurodevelopment may be potential candidates for schizophrenia. As an adaptor linking a number of tyrosine kinase receptors in multiple intracellular signaling cascades, Src homology 2 domain containing transforming protein 3 (SHC3) is a member of the Shc-like adaptor protein family, and expressed predominantly in the mature neurons of the central nervous system (CNS). In the present study, we aimed to investigate the association of SHC3 and schizophrenia. Methods An independent case-control association study was performed in a sample including 710 schizophrenia patients and 1314 healthy controls from a Northeast Chinese Han population. Results The allelic and genotypic association analyses showed that four SNPs in SHC3 significantly associated with schizophrenia(rs2316280, rs4877041, rs944485 and rs7021743). The haplotype composing of these four SNPs also showed significantly individual and global association with schizophrenia. Conclusion Our present results suggest SHC3 as a susceptibility gene for schizophrenia.

Modified Ferric Hydroxamate Spectrophotometry for Assaying Glycolic Acid from the Hydrolysis of Glycolonitrile by Rhodococcus sp. CCZU10-1

Yu-Cai He,You-Yan Liu,Cui-Luan Ma,Jian-He Xu 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.5

We successfully modified a ferric hydroxamate spectrophotometry method for assaying glycolic acid. Comparable to the high-performance liquid chromatography (HPLC)-based method, ferric hydroxamate spectrophotometry can be used to accurately monitor the time course of glycolonitrile bioconversion. Glycolic acid was assayed simply and rapidly at room temperature (25 ~35℃). Optimum culture conditions were obtained using this method to assay the glycolonitrile-hydrolyzing activity of Rhodococcus sp. CCZU10-1. The preferred carbon and nitrogen sources and ideal inducer were glucose (10 g/L),a composite of peptone (10 g/L) plus yeast extract (5 g/L),and ε-caprolactam (2 mmol/L), respectively. The optimal growth temperature and initial medium pH for Rhodococcus sp. CCZU10-1 glycolonitrile-hydrolyzing activity were 30℃ and pH 7.0. Modified ferric hydroxamate spectrophotometry could potentially be employed to assay other carboxylic acids.

ZnO Nanowire Arrays on 3D Hierachical Graphene Foam: Biomarker Detection of Parkinson’s Disease

Yue, Hong Yan,Huang, Shuo,Chang, Jian,Heo, Chaejeong,Yao, Fei,Adhikari, Subash,Gunes, Fethullah,Liu, Li Chun,Lee, Tae Hoon,Oh, Eung Seok,Li, Bing,Zhang, Jian Jiao,Huy, Ta Quang,Luan, Nguyen Van,Lee, Y American Chemical Society 2014 ACS NANO Vol.8 No.2

<P>We report that vertically aligned ZnO nanowire arrays (ZnO NWAs) were fabricated on 3D graphene foam (GF) and used to selectively detect uric acid (UA), dopamine (DA), and ascorbic acid (AA) by a differential pulse voltammetry method. The optimized ZnO NWA/GF electrode provided a high surface area and high selectivity with a detection limit of 1 nM for UA and DA. The high selectivity in the oxidation potential was explained by the gap difference between the lowest unoccupied and highest occupied molecular orbitals of a biomolecule for a set of given electrodes. This method was further used to detect UA levels in the serum of patients with Parkinson’s disease (PD). The UA level was 25% lower in PD patients than in healthy individuals. This finding strongly implies that UA can be used as a biomarker for PD.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancac3/2014/ancac3.2014.8.issue-2/nn405961p/production/images/medium/nn-2013-05961p_0005.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/nn405961p'>ACS Electronic Supporting Info</A></P>

A Novel 3D Polypseudo-rotaxane Metal-organic Framework Based on a Flexible Bis-pyridyl-bis-amide Ligand

Xiu-Li Wang,Na Han, Hong-Yan Lin,Chuang Xu,Jian Luan,Guo-Cheng Liu 대한화학회 2012 Bulletin of the Korean Chemical Society Vol.33 No.11

A novel 3D compound {[Cu(L)(H2O)4][Cu2(SIP)2(L)2]}·2H2O (1) (L = N,N-bis(4-pyridinecarboxamide)-1,4- butane, SIP = 5-sulfoisophthalate) is hydrothermally synthesized. X-ray diffraction analysis reveals that compound 1 is composed of 2D anionic [Cu2(SIP)2(L)2]n 2n− double-layers and discrete 1D cationic [CuL(H2O)4]n 2n+ polymeric chains, which represents a rare 3D polypseudo-rotaxane MOF from intercalation of 1D and 2D framework. In addition, the luminescent property and electrochemical behavior of compound 1 have been investigated.

A Novel 3D Polypseudo-rotaxane Metal-organic Framework Based on a Flexible Bis-pyridyl-bis-amide Ligand

Wang, Xiu-Li,Han, Na,Lin, Hong-Yan,Xu, Chuang,Luan, Jian,Liu, Guo-Cheng Korean Chemical Society 2012 Bulletin of the Korean Chemical Society Vol.33 No.11

A novel 3D compound $\{[Cu(L)(H_2O)_4][Cu_2(SIP)_2(L)_2]\}{\cdot}2H_2O$ (1) (L = N,N-bis(4-pyridinecarboxamide)-1,4-butane, SIP = 5-sulfoisophthalate) is hydrothermally synthesized. X-ray diffraction analysis reveals that compound 1 is composed of 2D anionic $[Cu_2(SIP)_2(L)_2]_n{^{2n-}}$ double-layers and discrete 1D cationic $[CuL(H_2O)_4]_n{^{2n+}}$ polymeric chains, which represents a rare 3D polypseudo-rotaxane MOF from intercalation of 1D and 2D framework. In addition, the luminescent property and electrochemical behavior of compound 1 have been investigated.