• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.132-137
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• 2010

The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

• Advances in nano research
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• v.6 no.2
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• pp.183-200
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• 2018

In the present study silver nanoparticles (AgNPs) were successfully synthesized using aqueous extract of Sargassum muticum. The aqueous extract (10%) treated with 1 mM silver nitrate solution resulted in the formation of AgNPs and the surface plasmon resonance (SPR) of the formed AgNPs was recorded at 360 nm using UV-Visible spectrophotometer. The molecules involved in the formation of AgNPs were identified by Fourier transform infrared spectroscopy (FT-IR), surface morphology was studied by using scanning electron microscopy (SEM), SEM micrograph clearly revealed the size of the AgNPs was in the range of 40-65 nm with spherical, hexagonal in shape and poly-dispersed nature, and X-ray diffraction spectroscopy (XRD) was used to determine the crystalline structure. High positive Zeta potential (36.5 mV) of formed AgNPs indicates the stability and XRD pattern revealed the crystal structure of the AgNPs by showing the Bragg's peaks corresponding to (111), (200), (311) and (222) planes of face-centered cubic crystal phase of silver. The synthesized AgNPs exhibited effective anticancerous activity (at doses 25 and $50{\mu}g/ml$ of AgNPs) against Breast cancer cell line (MCF7).

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1191-1196
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• 2009

The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.