[Objective] The purpose of this study was to clone a starch phosphorylase gene from Dunaliella salina and to preliminarily analyze its basic properties and protein expression. [Method] RT-PCR and RACE （rapid amplific...[Objective] The purpose of this study was to clone a starch phosphorylase gene from Dunaliella salina and to preliminarily analyze its basic properties and protein expression. [Method] RT-PCR and RACE （rapid amplification of cDNA ends） method was used for gene cloning; basic properties of the gene were analyzed using bioinformatics method; prokaryotic expression vector PGS21a-DsSP was constructed and transformed into E. coil BL21; the fusion protein was purified and detected by GST-SefinoseTM Kit and Western Blot, respectively. [Result] A starch phos-phorylase gene （GenBank accession No. KF061044） named DsSP was successfully isolated from D. salina. Basic properties, subcellular localization, secondary structure and tertiary structure of the protein were analyzed and predicted. The fusion protein was found in the supernatant and inclusion bodies. The supernatant protein was successfully purified. Western Blot analysis showed that the fusion protein was successfully expressed in E. coil BL21. [Conclusion] This study laid experimental foun- dation for further clarifying the function and mechanism of DsSP.展开更多
Acidogenic dissimilation of synthetic starch wastewater (1 000～10 000 mg COD·L -1 ) was studied in a thermophilic (55 ℃) upflow anaerobic sludge blanket (UASB) reactor.The production of volatile fatty acids...Acidogenic dissimilation of synthetic starch wastewater (1 000～10 000 mg COD·L -1 ) was studied in a thermophilic (55 ℃) upflow anaerobic sludge blanket (UASB) reactor.The production of volatile fatty acids (VFA) was proportional to the chemical oxygen demand (COD) loading rate.The yield of VFA was around 0.28 g VFA/g COD over the COD loading rate from 1.25 to 30 g COD·L -1 ·d -1 and the hydraulic retention time from 8.8 h to 24 h.Distribution of organic acids,the contents of propionic and butyric acids in the effluent in particular were also dependent on the COD loading rate.The thermophilic UASB reactor showed a stable performance on hydrolysis and acidogenesis of starch as well as suspended solid removal at short hydraulic retention times and high influent pH(10～11),during the operation of 110 d.展开更多
Magnetic starch particles （MSPs） were synthesized in water-in-oil mieroemulsion at room temperature. MSPs were characterized by transmission electron microscopy （TEM）, Fourier transform infrared spectrometry （FTI...Magnetic starch particles （MSPs） were synthesized in water-in-oil mieroemulsion at room temperature. MSPs were characterized by transmission electron microscopy （TEM）, Fourier transform infrared spectrometry （FTIR）, zeta potential system, thermogravimetric analysis （TGA） and vibrating sample magnetometry （VSM）. The average diameter of the MSPs was 220 nm, dispersed with well-proportioned size and magnetic resonance, the saturation magnetization was 3.64 A.mR/kg. MSP was coated with poly-L-lysine （PLL）, and then the surface of PLL-MSP was combined with fluorescein isothiocynate （FITC）. Results show that fluorescent/magnetic starch particles （FMSPs） are of stable photo-bleaching capability compared with free FITC, with low bio-toxicity and certain function of magnetic separation. It is expected that FMSPs are bifimctional nano-materials including fluorescence labelling and magnetic separation.展开更多
基金Supported by National Natural Science Foundation of China(No.30972240)Science and Technology Project of Liaoning Provincial Department of Education(No.2008T023)~~
文摘[Objective] The purpose of this study was to clone a starch phosphorylase gene from Dunaliella salina and to preliminarily analyze its basic properties and protein expression. [Method] RT-PCR and RACE （rapid amplification of cDNA ends） method was used for gene cloning; basic properties of the gene were analyzed using bioinformatics method; prokaryotic expression vector PGS21a-DsSP was constructed and transformed into E. coil BL21; the fusion protein was purified and detected by GST-SefinoseTM Kit and Western Blot, respectively. [Result] A starch phos-phorylase gene （GenBank accession No. KF061044） named DsSP was successfully isolated from D. salina. Basic properties, subcellular localization, secondary structure and tertiary structure of the protein were analyzed and predicted. The fusion protein was found in the supernatant and inclusion bodies. The supernatant protein was successfully purified. Western Blot analysis showed that the fusion protein was successfully expressed in E. coil BL21. [Conclusion] This study laid experimental foun- dation for further clarifying the function and mechanism of DsSP.
文摘Acidogenic dissimilation of synthetic starch wastewater (1 000～10 000 mg COD·L -1 ) was studied in a thermophilic (55 ℃) upflow anaerobic sludge blanket (UASB) reactor.The production of volatile fatty acids (VFA) was proportional to the chemical oxygen demand (COD) loading rate.The yield of VFA was around 0.28 g VFA/g COD over the COD loading rate from 1.25 to 30 g COD·L -1 ·d -1 and the hydraulic retention time from 8.8 h to 24 h.Distribution of organic acids,the contents of propionic and butyric acids in the effluent in particular were also dependent on the COD loading rate.The thermophilic UASB reactor showed a stable performance on hydrolysis and acidogenesis of starch as well as suspended solid removal at short hydraulic retention times and high influent pH(10～11),during the operation of 110 d.
基金Project(200501) supported by the "985" Program of China
文摘Magnetic starch particles （MSPs） were synthesized in water-in-oil mieroemulsion at room temperature. MSPs were characterized by transmission electron microscopy （TEM）, Fourier transform infrared spectrometry （FTIR）, zeta potential system, thermogravimetric analysis （TGA） and vibrating sample magnetometry （VSM）. The average diameter of the MSPs was 220 nm, dispersed with well-proportioned size and magnetic resonance, the saturation magnetization was 3.64 A.mR/kg. MSP was coated with poly-L-lysine （PLL）, and then the surface of PLL-MSP was combined with fluorescein isothiocynate （FITC）. Results show that fluorescent/magnetic starch particles （FMSPs） are of stable photo-bleaching capability compared with free FITC, with low bio-toxicity and certain function of magnetic separation. It is expected that FMSPs are bifimctional nano-materials including fluorescence labelling and magnetic separation.
