Jiangsu Hanbon Science and Technology Co.

Huaian, China

Jiangsu Hanbon Science and Technology Co.

Huaian, China
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Wang Z.,Jiangnan University | Cai Y.,Jiangnan University | Liao X.,Jiangnan University | Zhang F.,Jiangnan University | And 2 more authors.
Applied Biochemistry and Biotechnology | Year: 2010

A new white-rot fungus SYBC-L1, which could produce an extracellular laccase, was isolated from a decayed Elaeocarpus sylvestris. The strain was identified as Pycnoporus sp. SYBC-L1 according to the morphological characteristics and ribosomal ITS1-5.8S-ITS2 RNA genomic sequence analysis. The highest laccase activity of 24.1 Uml-1, which was approximately 40-fold than that in basal medium, was achieved in optimal culture medium in submerged fermentation. The laccase produced by Pycnoporus sp. SYBC-L1 was not only a cold adaptation enzyme with a relative catalytic activity of 30.2% at 0°C but also a high thermostable enzyme. The half-lives at 60, 70 and 80°C were 85.5, 37.2, and 2.6 h, respectively. The laccase could effectively decolorize weak acid blue AS and diamond black PV up to 88% and 74.7%, respectively, within 2 h in the absence of any redox mediators. The results suggested Pycnoporus sp. SYBC-L1 was a potential candidate for laccase production and industrial application. © 2009 Humana Press.

Cai Y.,Jiangnan University | Wu H.,Jiangnan University | Liao X.,Jiangnan University | Ding Y.,Jiangnan University | And 2 more authors.
Biotechnology and Bioprocess Engineering | Year: 2010

A novel manganese peroxidase of Rhizoctonia sp. SYBC-M3 (R-MnP) was purified by (NH4)2SO4 fractionation, DEAE-cellulose-32 column chromatography, and Sephadex G100 column chromatography. The molecular mass of R-MnP was determined to be approximately 40.4 kDa by SDS-PAGE. The optimum temperature and pH for R-MnP were 55°C and 4.5, respectively. R-MnP was highly stability when the temperature was below 50°C. R-MnP could retain about 60% of its activity when the pH was between 4 and 6.5. However, R-MnP activity was inhibited by Fe3+, Cu 2+, and Co3+ as well as increased by Zn2+ and Ca2+. R-MnP demonstrated oxidation of DMP, ABTS, veratryl alcohol, and guaiacol. The Km values of RMnP for H2O2 and Mn2+ were 25.3 and 53.9 μmol/L, respectively. © 2010 The Korean Society for Biotechnology and Bioengineering and Springer-Verlag.

Wang Z.-X.,Jiangnan University | Wang Z.-X.,Hebei University of Science and Technology | Cai Y.-J.,Jiangnan University | Liao X.-R.,Jiangnan University | And 4 more authors.
Process Biochemistry | Year: 2010

The white-rot fungus Pycnoporus sp. SYBC-L1 produced large amount of laccase in submerged fermentation. Two laccase isozymes (LacI and LacII) were purified using (NH4)2SO4 fractionation, DEAE-cellulose and Sephadex G-100 column chromatography. The molecular masses of LacI and Lac II were 55.89 and 63.07kDa, respectively by SDS-PAGE. Both the laccases showed acidic pH optima and high catalytic activities at low temperature for oxidations of 2,6-dimethoxyphenol (DMP), 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonate acid) (ABTS), syringaldazine and guaiacol. LacI and LacII were not only with high cold adaptation, but also fairly stable at high temperature. The half-lives of LacI at 50, 60 and 70°C were 69.31, 2.58 and 0.13h, respectively, whereas LacII was more stable with half-lives of 256.72, 21.00 and 2.06h respectively. The best substrates for the enzymes were both found to be ABTS, in which the Km values of LacI and LacII were 0.0166 and 0.0435mM and the catalytic efficiencies were 19640.36 and 31172.64S-1mM-1, respectively. EDTA and low concentration of Cu2+ and Mn2+ almost had non-inhibitions on their activities. LacII with syringaldehyde efficiently decolorized Remazol Brilliant Blue R. The high thermostabilities as well as cold adapted properties made Pycnoporus sp. SYBC-L1 laccases to be excellent candidates in harsh industry. © 2010 Elsevier Ltd.

Yuan B.-H.,Jiangnan University | Cai Y.-J.,Jiangnan University | Liao X.-R.,Jiangnan University | Yun L.-H.,Jiangnan University | And 2 more authors.
African Journal of Biotechnology | Year: 2010

A cold-adapted lipase producing strain of mesophilic bacterium, named SYBC LIP-Y, was isolated from the decayed seeds of Ginkgo biloba L. by screening with plates containing Victoria Blue B and with the flask-shaking fermentation. It was identified as a novel Burkholderia species. The properties of its lipase after purification by PEG1000/ potassium phosphate aqueous two-phase system were characterized. The optimal pH and temperature of the enzyme was 10.0 and 30°C, respectively. About 80% of the original activity is maintained by heating at 40°C for 60 min. The lipase could also retained 70% of the maximal activity at the temperature of 0°C and suggested that it may be belonged to the cold-adapted lipase. Thus, it was proved to have good temperature stability and might have wide applicating fields. © 2010 Academic Journals.

Liu J.,Jiangnan University | Liu J.,University of Georgia | Cai Y.,Jiangnan University | Liao X.,Jiangnan University | And 5 more authors.
Journal of Cleaner Production | Year: 2013

Laccase is one of the most useful green enzymes for cleaner industrial application and environmental protection. It is necessary to produce laccase with known composition of its commercial style at low production cost in an efficient way. In this study, the efficiency of laccase production by Pycnoporus sp. SYBC-L3 in a 65-L air-lift reactor was evaluated and the main extracellular proteins were identified. The highest laccase activity approximated 72,000 U/L after 6 days' cultivation. Using nano-LC-MS/MS protein identification technology, 22 different peptide fragments were identified matching sequences in different proteins with laccase as the predominant extracellular one. The crude laccase exhibited high catalytic activity at temperatures ranging from 0 °C to 100 °C at an acidic condition, with respective relative activity of 40% at 0 °C and 80% at 100 °C. The crude laccase was found to be very thermal stable, with half-lives of 4 h, 6.5 h and 10 h at 70 °C, 60 °C and 50 °C, respectively. The crude laccase still retained approximately 85% of its initial activity after one year of storage at room temperature. These results showed that laccase can be effectively produced by Pycnoporus sp. SYBC-L3 in air-lift reactor at larger scale and has great potential for industrial production and applications. © 2012 Elsevier Ltd. All rights reserved.

Liu J.,Key Laboratory of Industrial Biotechnology | Liu J.,University of Georgia | Cai Y.,Key Laboratory of Industrial Biotechnology | Liao X.,Key Laboratory of Industrial Biotechnology | And 4 more authors.
Bulletin of Environmental Contamination and Toxicology | Year: 2012

We conducted experiments to culture Pycnoporus sp. SYBC-L3 in a medium comprising an industrial waste (dye-containing textile effluent) and a lignocellulosic waste (Phragmites australis) that achieved laccase production while having the color removed from the wastewater. Our experimental results showed that the fungus grew well in liquid submerged cultivation with the diluted textile effluent as the sole culture medium, but relatively low extracellular laccase activity (1.8 U/mL) was produced. Addition of the lignocellulosic biomass enhanced laccase production and color removal. The highest laccase activity was found to be 6.5 U/mL in the presence of Phragmites australis stem. Under this condition, 70 % color removal occurred in the culture medium. This study provided an alternative novel scheme to remove color in textile wastewater while having an economic value added by producing laccase. © Springer Science+Business Media, LLC 2012.

Cai Y.,Jiangnan University | Liao X.,Jiangnan University | Liang X.,Jiangnan University | Ding Y.,Jiangnan University | And 2 more authors.
New Biotechnology | Year: 2011

Hypocrellins are important photodynamic therapy compounds for cancer disease. The effect of surfactants on hypocrellin production of Shiraia sp. SUPER-H168 was evaluated under submerged fermentation condition. The production of hypocrellins could reach 780.6. mg/l with the addition of Triton X-100, confirmed by color reaction, high performance liquid chromatography, electrospray ionization mass spectrometry and nuclear magnetic resonance experiments. According to our observation, treatment of the culture at the beginning of the fermentation was most effective, and the yield of hypocrellins was much lower with the addition of Triton X-100 during the log phase and stationary phase. Shiraia sp. SUPER-H168 could not produce hypocrellin with the addition of other tested surfactants, such as Tween 40, Triton X-114 and SDS. The experimental results indicated that Shiraia sp. SUPER-H168 could not produce hypocrellins without Triton X-100 under submerged fermentation condition. © 2011 Elsevier B.V.

Cai Y.,Jiangnan University | Liu S.,Jiangnan University | Liao X.,Jiangnan University | Ding Y.,Jiangnan University | And 2 more authors.
Food and Bioproducts Processing | Year: 2011

Three AFPs (AFP-1, AFP-2, AFP-3) of Ligustrum lucidum Ait leaves were purified by using ice-affinity, chromatography separation on a DEAE-cellulose-32 column and a Sephadex G100 column. The ice-affinity proteins were about 1.2 mg/100 g leaves. Their molecular weight was 66.1, 26.3, and 20.2 kDa, respectively. Their thermal hysteresis activity was 0.379, 0.678, and 0.460 °C respectively. Asx was very abundant in these AFPs; its molar ratio was 22.2, 24.9, and 27.8%, respectively. AFP-2 was effective to protect peroxidase, β-glucosidase, and trehalose synthase from freeze-thawing process. Cryoprotection of AFP-2 is better than trehalose. © 2010 The Institution of Chemical Engineers.

Tang S.,CAS Lanzhou Institute of Chemical Physics | Tang S.,University of Chinese Academy of Sciences | Wang L.,CAS Lanzhou Institute of Chemical Physics | Han H.,Jiangsu Hanbon Science and Technology CO. | And 3 more authors.
RSC Advances | Year: 2013

A new mixed-mode organic-silica hybrid monolithic column for capillary electrochromatography was synthesized via a sol-gel process and a following post-modification with 4,4′-dipyridine. Characterization by SEM shows that the hybrid monolith has homogenous macroporous morphology and it is well attached to the inner wall of the capillary. A stable and reversed electroosmotic flow (EOF) was generated at acidic pH due to the pyridinium groups on the surface of the stationary phase. The column performance was evaluated by separating various kinds of compounds, such as polycyclic aromatic hydrocarbons (PAHs), alkylbenzenes, phenols, inorganic anions and organic acids. Multiple retention mechanisms including hydrophobic, π-π and anion-exchange interactions were exhibited at separation of the analytes. The monolithic stationary phase exhibited reversed-phase chromatographic behavior toward neutral solutes. Meanwhile, inorganic anions and organic acids can be separated by the mixed-mode mechanism comprising electrophoresis, hydrophobic and anion-exchange interactions. Good performance and repeatability showed that this new hybrid column can be used in the analysis of various compounds with great prospects. © 2013 The Royal Society of Chemistry.

Xie R.,China Pharmaceutical University | Xie R.,Shanghai Key Laboratory for Pharmaceutical Metabolite Research | Xie R.,Shanghai Research Center for Drug Chinese Materia Medica Metabolism | Wen J.,China Pharmaceutical University | And 9 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2010

An automated system using on-line solid-phase extraction and HPLC with UV detection was developed for the determination of faropenem in human plasma and urine. Analytical process was performed isocratically with two reversed-phase columns connected by a switching valve. After simple pretreatment for plasma and urine with acetonitrile, a volume of 100 μl upper layer of the plasma or urine samples was injected for on-line SPE column switching HPLC-UV analysis. The analytes were retained on the self-made trap column (Lichrospher C18, 4.6 mm × 37 mm, 25 μm) with the loading solvent (20 mM NaH2PO4 adjusted pH 3.5) at flow rate of 2 ml min-1, and most matrix materials were removed from the column to waste. After 0.5 min washing, the valve was switched to another position so that the target analytes could be eluted from trap column to analytical column in the back-flush mode by the mobile phase (acetonitrile-20 mM NaH2PO4 adjusted pH 3.5, 16:84, v/v) at flow rate of 1.5 ml min-1, and then separated on the analytical column (Ultimate™ XB-C18, 4.6 mm × 50 mm, 5 μm).The complete cycle of the on-line SPE preconcentration purification and HPLC separation of the analytes was 5 min. Calibration curves with good linearities (r = 0.9994 for plasma sample and r = 0.9988 for urine sample) were obtained in the range 0.02-5 μg ml-1 in plasma and 0.05-10 μg ml-1 in urine for faropenem. The optimized method showed good performance in terms of specificity, linearity, detection and quantification limits, precision and accuracy. The method was successfully utilized to quantify faropenem in human plasma and urine to support the clinical pharmacokinetic studies. © 2009 Elsevier B.V. All rights reserved.

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