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Zhu D.F.,Ningbo University | Hu Z.H.,Ningbo University | Hu Z.H.,Marine Fishery Research Institute of Zhejiang Province | Shen J.M.,Ningbo University
Crustaceana | Year: 2011

In the present study, a genomic DNA of MIH (GenBank: #EU869539) was cloned from the swimming crab, Portunus trituberculatus (Miers, 1876). The genome DNA, consisting of 2865 bp, is comprised of three exons interrupted by two introns. Multiple sequence alignments revealed that in the 5′ upstream region of MIH, sequences with high similarity to arthropod initiator, TATA box, CREB (cyclic AMP response element binding) protein were the common structure. The signal peptide in the genomic DNA was encoded by exon1 and exon2, which was interrupted by 242 bp-intron (intron1), located between gln 12 and arg 13. The mature peptide was encoded by exon2 and exon3, which was interrupted by 313 bp-intron (intron2), at the position between 2 nd and 3 rd nucleotides of the codon encoding arg 41. Pot-MIH was expressed only in the eyestalk ganglia, ovaries, testes, posterior spermatic duct, bristle in ejaculatory duct, cranial ganglia, and thoracic ganglia, as determined in various tissues by semi-quantitative RT-PCR. A cDNA encoding the mature peptide was used to express recombinant MIH (rMIH) using the Escherichia coli (Migula, 1895) expression system. Two constructs were designed to yield either a mature MIH fusion protein with and without histidine (His) tag at the carboxyl terminus. The rMIH protein was detected by SDS-PAGE and Western blot analysis, indicating that the antibody prepared by two rMIH proteins has high specificity. © 2011 BRILL. Source

Gao Y.,Zhejiang Ocean University | Jin L.,Zhejiang Ocean University | Jin L.,Marine Fishery Research Institute of Zhejiang Province | Shi H.,Marine Fishery Research Institute of Zhejiang Province | Chu Z.,Zhejiang Ocean University
Journal of Agricultural and Food Chemistry | Year: 2015

Bacillus sp. strain hys-1, which was isolated from active sludge, could degrade >90% butachlor at a concentration of 100 mg/L within 7 days. The present work revealed that strain hys-1 could mineralize butachlor via the following pathway: butachlor was initially metabolized to 2-chloro-N-(2,6-diethylphenyl)-N-methylacetamide by debutoxylation and then transformed to form 2-chloro-N-(2,6-diethylphenyl)acetamide by N-demethylation. Subsequently, it was converted to 2,6-diethylaniline and further mineralized into CO2 and H2O. In addition, the catalytic efficiency of crude cell extracts descended as follows: alachlor > acetochlor > butachlor. Furthermore, a novel 744 bp gene responsible for transforming butachlor into 2-chloro-N-(2,6-diethylphenyl)-N-methylacetamide was cloned from strain hys-1 and the encoding debutoxylase was designated Dbo. Then Dbo was expressed in Escherichia coli BL21 (DE3) and purified using Ni-nitrilotriacetic acid affinity chromatography. Dbo displayed the highest activity against butachlor at pH 6.5 and 30 °C. Metal ions played an important role in Dbo activity. To the best of the authors knowledge, this is the first report that strain hys-1 can mineralize butachlor by a novel metabolic mechanism and the first identification of a gene encoding butachlor debutoxylase. © 2015 American Chemical Society. Source

Feng M.,Marine Fishery Research Institute of Zhejiang Province | Feng M.,Zhejiang Ocean University | Zhang W.,CAS Qingdao Institute of Oceanology | Wang W.,Marine Fishery Research Institute of Zhejiang Province | And 4 more authors.
Marine Pollution Bulletin | Year: 2015

Ciliated protozoa have many advantages in bioassessment of water quality. The ability of tintinnids for assessing water quality status was studied during a 7-year. cycle in Jiaozhou Bay of the Yellow Sea, northern China. The samples were collected monthly at four sites with a spatial gradient of environmental pollution. Environmental variables, e.g., temperature, salinity, chlorophyll a (Chl a), dissolved inorganic nitrogen, soluble reactive phosphate (SRP), and soluble active silicate (SRSi), were measured synchronously for comparison with biotic parameters. Results showed that: (1) tintinnid community structures represented significant differences among the four sampling sites; (2) spatial patterns of the tintinnid communities were significantly correlated with environmental variables, especially SRSi and nutrients; and (3) the community structural parameters and the five dominant species were significantly correlated with SRSi and nutrients. We suggested that tintinnids may be used as a potential bioindicator for discriminating water quality status in marine ecosystems. © 2015 Elsevier Ltd. Source

Mei G.-M.,Marine Fishery Research Institute of Zhejiang Province | Hao Q.,Beijing Haiyan Pharmaceutical Co. | Zhang X.-J.,Marine Fishery Research Institute of Zhejiang Province | Guo Y.-M.,Marine Fishery Research Institute of Zhejiang Province | Chen X.-C.,Marine Fishery Research Institute of Zhejiang Province
Modern Food Science and Technology | Year: 2014

A new polysaccharide SP2 was isolated from Lentinusedodes using an aqueous HCl solution and purified using gel column chromatography, the purity was determined using filter paper electrophoresis. The relative molecular mass and distribution of SP2 were analyzed by high-performance gel permeation chromatography, and the structure was analyzed by UV-Vis spectroscopy, gas chromatography, infrared spectroscopy, X-ray diffraction, and differential scanning calorimetry, as well as periodate oxidation, Smith degradation, and Congo red experiments. The results indicated that SP2 showed a typical absorption peak for polysaccharides after the color reaction and had a relatively homogeneous composition. In addition, the molecular weight distribution range was relatively narrow, with a weight-average molecular weight of 5.203×104 u. SP2 was found to be composed of mannose, glucose, and galactose with a molar ratio of 0.32:58.6:2.82. Periodate oxidation and Smith degradation experiments indicated that the C-C bonds in the branches of SP2 were composed of (1→2), (1→3), and (1→6) linkages with a molar ratio of 9.43:5.44:1. SP2 was found to be a glycoprotein conjugate containing pyranose rings that was amorphous and could not form the monocry stalline structure. SP2 contained spiral structures and had no strong intermolecular interactions. These results provided evidence to explain the structure-activity relationships of polysaccharides from L. edodes. Source

Jin L.,Marine Fishery Research Institute of Zhejiang Province | Zhang X.,Marine Fishery Research Institute of Zhejiang Province | Sun X.,Marine Fishery Research Institute of Zhejiang Province | Shi H.,Marine Fishery Research Institute of Zhejiang Province | Li T.,Marine Fishery Research Institute of Zhejiang Province
World Journal of Microbiology and Biotechnology | Year: 2014

A strain, designated as FM-6, was isolated from fish. Based on the results of phenotypic, physiological characteristics, genotypic and phylogenetic analysis, strain FM-6 was finally identified as Paenibacillus sp. When albendazole was provided as the sole carbon source, strain FM-6 could grow and transform albendazole. About 82.7 % albendazole (50 mg/L) was transformed by strain FM-6 after 5 days incubation at 30 °C, 160 rpm. With HPLC–MS method, the transforming product of albendazole was researched. Based on the molecular weight and the retention time, product was identified as albendazole sulfoxide and the transforming pathway of albendazole by strain FM-6 was proposed finally. The optimum temperature and pH for the bacterium growth and albendazole transformation by strain FM-6 were both 30 °C and 7.0. Moreover, the optimum concentration of albendazole for the bacterium growth was 50 mg/L. Coupled with practical production, 50 mg/L was the optimum concentration of albendazole transformation for strain FM-6. This study highlights an important potential use of strain FM-6 for producing albendazole sulfoxide. © 2014, Springer Science+Business Media Dordrecht. Source

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