Lin Xiao

Nanjing University

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Liuyan Yang

Shanghai Academy of Agricultural Sciences

Yuke Peng

University of Shanghai for Science and Technology

Lijuan Jiang

Central South University

Liuyan Yang

State Key Laboratory of Pollution Control and Resource Reuse

Ai‐Jun Miao

State Key Laboratory of Pollution Control and Resource Reuse

Huiping Chen

South China Agricultural University

Yuqiong Tang

State Key Laboratory of Pollution Control and Resource Reuse

Dongmei Yue

Liaoning Ocean and Fisheries Research Institute

Hongwei Du

University of Science and Technology of China

Junling Lu

Shaanxi Normal University

Cooperative Institutions

Nanjing University

State Key Laboratory of Pollution Control and Resource Reuse

Chinese Academy of Sciences

University of Chinese Academy of Sciences

Sun Yat-sen University

Peking University

Tongji University

Huazhong University of Science and Technology

Sichuan University

Jinan University

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Characterization of depth-related microbial communities in lake sediment by denaturing gradient gel electrophoresis of amplified 16S rRNA fragments

Journal of Environmental Sciences (2008)

Xingqing Zhao Liuyan Yang Zhenyang Yu Naiying PENG Lin Xiao

Characterization

10.1016/s1001-0742(08)60035-2

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Citations (50)

[Integration and expression of polyphosphate kinase gene in Pseudomonas putida].

PubMed (2009)

Hongwei Du Jun Wu Lin Xiao Liuyan Yang Lijuan Jiang

In order to construct high accumulating-phosphate microorganism, the ppk gene from E. coli was inserted to the broad-host-range plasmid pBBR1MCS-2 to form plasmid pBBR1MCS-2-ppk. The complete ppk gene with promoter and terminator sequences from pBBR1MCS-2-ppk was then cloned and inserted to suicide plasmid pUTmini-Tn5 to form plasmid pUTmini-Tn5-ppk, which was transformed into Pseudomonas putida KT2440 by triparental conjugation. Finally, ppk gene was integrated into the chromosomal DNA of KT2440. The results of RT-PCR showed that the selected genetically engineered bacterium KT2440-PPK expressed ppk efficiently, while KT2440 as control expressed weakly. The results of artificial wastewater treatment showed after 1h inoculation, the concentration of poly-phosphate in KT2440-PPK came to the maximum approximately 3.05 mg/g, which was 15 times higher than that in KT2440 at the same experimental condition. And KT2440-PPK can remove more than 90% phosphate in artificial wastewater.

Pseudomonas putida

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[Enhanced cellulase production of Penicillium decumbens by knocking out CreB encoding a deubiquitination enzyme].

PubMed (2012)

Guangqi Zhou Jing Lü Zhonghai Li Jingjing Li Mingyu Wang

Penicillium decumbens T. is an important filamentous fungus for the production of cellulases to effectively degrade lignocellulose for second generation biofuel production. In order to enhance the capability of Penicillium decumbens to produce cellulases, we constructed a creB (a deubiquitinating enzyme encoding gene) deletion cassette, and generated a creB knockout strain with homologous double crossover recombination. This mutation resulted in a detectable decrease of carbon catabolite repression (CCR) effect. The filter paper activity, endoglucanase activity, xylanase activity and exoglucanase activity of the deltacreB strain increased by 1.8, 1.71, 2.06 and 2.04 fold, respectively, when comparing with the parent strain Ku-39. A 2.68 fold increase of extracellular protein concentration was also observed. These results suggest that the deletion of creB results in CCR derepression. These data also suggest that CREB influences cellulase production of Penicillium decumbens. In generation, this study provides information that can be helpful for constructing cellulase hyper-producing strain.

Derepression

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Cadmium Toxicity to Microcystis aeruginosa PCC 7806 and Its Microcystin-Lacking Mutant

PLoS ONE (2015)

Bin Huang Shen Xu Ai‐Jun Miao Lin Xiao Liuyan Yang

The adverse effects of microcystin (MC) produced by cyanobacteria have drawn considerable attention from the public. Yet it remains unclear whether MC confers any benefits to the cyanobacteria themselves. One suggested function of MC is complexation, which may influence the bioaccumulation and toxicity of trace metals. To test this hypothesis, we examined Cd toxicity to wild-type Microcystis aeruginosa PCC 7806 (WT) and its MC-lacking mutant (MT) under nutrient-enriched (+NP), phosphorus-limited (-P), and nitrogen-limited (-N) conditions. The accumulation of Cd and the biochemical parameters associated with its detoxification [total phosphorus (TP), inorganic polyphosphate (Poly-P), and glutathione (GSH) in the cells as well as intra- and extra-cellular carbohydrates] were quantified. Although the –P cyanobacteria accumulated less Cd than their +NP and –N counterparts, the different nutrient-conditioned cyanobacteria were similarly inhibited by similar free ion concentration of Cd in the medium ([Cd2+]F). Such good toxicity predictability of [Cd2+]F was ascribed to the synchronous decrease in the intracellular concentrations of Cd and TP. Nevertheless, Cd toxicity was still determined by the intracellular Cd to phosphorus ratio (Cd/P), in accordance with what has been reported in the literature. On the other hand, the concentrations of TP, Poly-P, and carbohydrates went up, but GSH concentration dropped down with the enhancement of [Cd2+]F, indicating their association with Cd detoxification. Although the inactivation of MC peptide synthetase gene had some nutrient and Cd concentration dependent effects on the parameters above, both cyanobacterial strains showed the same Cd accumulation ability and displayed similar Cd sensitivity. These results suggest that MC cannot affect metal toxicity either by regulating metal accumulation or by altering the detoxification ability of the cyanobacteria. Other possible functions of MC need to be further investigated.

Microcystis aeruginosa

Microcystis

Microcystin-LR

10.1371/journal.pone.0116659

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Citations (14)

Calculating Methods of Heat Transfer Quantity through Surrounding Rocks in Metro Design

Chinese Journal of Underground Space and Engineering (2008)

Lin Xiao

The heat transfer quantity through surrounding rocks is a basic element in ventilation and conditioning design of the metro.At present,it is difficult to determine this parameter and a uniform standard hasn't been formed.In this paper,based on an infinite tunnel model,a formula is deduced by considering the heat transfer of the station and tunnel synthetically,and it is compared with some related methods in a real example.The law of heat transfer is analyzed by numerical simulation,and it is found that there are large discrepancies among these different methods and the heat transfer quantity through surrounding rocks in metro design is more than the result derived from the formula.

Transfer station

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Bacterioplankton community structure in a eutrophic lake in relation to water chemistry

World Journal of Microbiology and Biotechnology (2009)

Jin Zeng Liuyan Yang Hongwei Du Lin Xiao Lijuan Jiang

Bacterioplankton

Canonical correspondence analysis

Biogeochemical Cycle

Anammox

10.1007/s11274-008-9946-5

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Citations (28)

Purification of discharged residual guar fracturing fluid

Shiyou Zuancai Gongyi (2016)

Lin Xiao Yao Yuanyuan Chen Guo

Fracturing fluid

Residual oil

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Expression of glutathione S-transferase fusion proteins of human CYP1A1 and preparation of anti CYP1A1 polyclonal antibody

Zhongguo yaolixue yu dulixue zazhi (2000)

Lin Xiao

Our aim is to get high amounts of pure antigens to raise specific antibodies and to perform quantifications. Fusion proteins constructed between glutathione S transferase(GST) and CYP1A1 was expressed in Escherichia coli DH5α. We identified recombinants pGEX/1A1 by direct expression screening. Insoluble proteins were isolated from the bacteria and the fusion proteins were purified from a preparative (2 mm) SDS PAGE. The polyacrylamide gel containing the fusion proteins GST 1A1 was used to immunize BALB/c mice from which polyclonal ascites fluid was prepared. Anti CYP1A1 polyclonal antibodies(1A1pAb) was purified by cross absorption with GST 2B6(GST fusion protein of CYP2B6 204-353 amino acid expressed in this laboratory, purified by preparative SDS PAGE)and then by passing through protein A Sepharose affinity column. The purified GST 2B6 and GST 1A1 were used to test the specificity of 1A1pAb. 1A1pAb is specific to GST 1A1 and is still weakly crossreactive with GST 2B6. But CYP1A1 can be discriminated from other CYP subtypes.

Polyclonal antibodies

Glutathione S-transferase

Sepharose

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Enhancing butanol tolerance of Escherichia coli reveals hydrophobic interaction of multi-tasking chaperone SecB

Biotechnology for Biofuels (2019)

Guochao Xu Anning Wu Lin Xiao Ruizhi Han Ye Ni

Escherichia coli has been proved to be one promising platform chassis for the production of various natural products, such as biofuels. Product toxicity is one of the main bottlenecks for achieving maximum production of biofuels. Host strain engineering is an effective approach to alleviate solvent toxicity issue in fermentation.Thirty chaperones were overexpressed in E. coli JM109, and SecB recombinant strain was identified with the highest n-butanol tolerance. The tolerance (T) of E. coli overexpressing SecB, calculated by growth difference in the presence and absence of solvents, was determined to be 9.13% at 1.2% (v/v) butanol, which was 3.2-fold of the control strain. Random mutagenesis of SecB was implemented and homologously overexpressed in E. coli, and mutant SecBT10A was identified from 2800 variants rendering E. coli the highest butanol tolerance. Saturation mutagenesis on T10 site revealed that hydrophobic residues were required for high butanol tolerance of E. coli. Compared with wild-type (WT) SecB, the T of SecBT10A strain was further increased from 9.14 to 14.4% at 1.2% butanol, which was 5.3-fold of control strain. Remarkably, E. coli engineered with SecBT10A could tolerate as high as 1.8% butanol (~ 14.58 g/L). The binding affinity of SecBT10A toward model substrate unfolded maltose binding protein (preMBP) was 11.9-fold of that of WT SecB as determined by isothermal titration calorimetry. Residue T10 locates at the entrance of hydrophobic substrate binding groove of SecB, and might play an important role in recognition and binding of cargo proteins.SecB chaperone was identified by chaperone mining to be effective in enhancing butanol tolerance of E. coli. Maximum butanol tolerance of E. coli could reach 1.6% and 1.8% butanol by engineering single gene of SecB or SecBT10A. Hydrophobic interaction is vital for enhanced binding affinity between SecB and cargo proteins, and therefore improved butanol tolerance.

Saturated mutagenesis

Protein Engineering

Thermostability

Metabolic Engineering

Isothermal Titration Calorimetry

10.1186/s13068-019-1507-7

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Citations (14)

Function of TTG1A in hepatic stellate cells

Lin Xiao Jun Cheng Liying Zhang Jiang Guo Hong Yuan

Objective To investigate the biological functions of TTG1A in liver fibrosis.Methods Yeast two-hybrid system Was,used to screell proteins associated with TTG1A.Briefly,the coding sequence of TTG1A was cloned into Pgbkt7 vector,and the recombinant plasmid was transformed into yeast cells AH109(a type),then these cells were mated with yeast cells Y187(αtype)transformed with human leukocyte cDNA library plasmid pACT2.The obtained diploid yeast cells were plated on synthetic dropout nutrient meditim containing X-a-gal for double selection.The plasmids from positive colonies were transformed into E.coli and sequenced.Results The recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed.Nineteen TTG1A binding proteins,including homo sapiens major histocompatibilitycomplex,class Ⅱ DP beta 1(HLA-DPβ1),homo sapiens ribosomal protein L30(RPL30),homo sapiens nuclcophosmin homo sapiens nucleobindin 2(NUCB2),homo sapiens ash2,variant Gaucher disease and variant memchromatic leukodystrophy,MORF4L1,homo sapiens ubiquitin-coujugating enzyme E2L3(UBE2L3),APOA1,homo sapiens lectin,and galectin 1,were idedtified.Conclusions This study may help to elucidate the molecular function of TTG1A. Key words: Transforming growth factor beta; Hepatic stellate cells; Liver cirrhosis; Gene; TrGlA

Homo sapiens

10.3760/cma.j.issn.1007-3418.2009.12.010

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