Effects of Temperature on Determination of Protein Concentration with Coomassie Brilliant Blue Method

[Objective] This study aimed to investigate the effects of temperature on determination of protein concentration with Coomassie Brilliant Blue method,thus proving advice and guidance for accurate determination of protein concentration.[Method] With Coomassie Brilliant Blue method,the concentrations of different bovine serum albumin samples were determined under different temperatures and incubation time.[Result] According to the standard curve,when the determination range of protein concentration was 0-100 mg/ml,the determined protein concentration was relatively stable after incubation at 20 ℃ for 20-30 min.Furthermore,the determination result of higher protein concentration with Coomassie Brilliant Blue method was less affected by various factors.[Conclusion] In determination of protein concentration with Coomassie Brilliant Blue method,temperature,sample concentration and incubation time were important factors affecting the accuracy of experimental results.

Biodegradation and detoxification of the triphenylmethane dye coomassie brilliant blue by the extracellular enzymes from mycelia of Lactarius deliciosus

Fungi play an important role in dying wastewater treatment.In this work,the mycelia of Lactarius deliciosus exhibited an excellent capacity in decolorizing coomassie brilliant blue(CBB).The results demonstrated that the mycelia could treat CBB with high concentrations over a broad range of pH and temperature.The decolorization rate of 99.19%and the removal rate of 16.31 mg·L^(‒1)·h were realized.The mycelia could be recycled from decolorizing process for 19 times,indicating a good re-usability.It verified that the lignin peroxidase(121.65 U·L^(‒1))and manganese peroxidase(36.77 U·L^(‒1))were involved in the degradation and decolorization process of CBB.Toxicity assessments indicated the seed germination rate was up to 82.22%while inhibition to Escherichia coli decreased dramatically and no significant effect on Caenorhabditis elegans growth was found.The removal of CBB was a synergistic process accomplished by adsorption and biodegradation.The mycelia could be used for eco-friendly CBB treatment.

Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.

Micellar sensitized catalytic kinetic spectrophotometry for highly accurate and reproducible determination of Ⅴ(Ⅳ) and Ⅴ(Ⅴ)

A highly accurate and reproducible micellar sensitized kinetic method was proposed for determination of V（VI）. The method is based on its catalytic effect on the oxidation of Coomassie brilliant blue R 250 （CBB＋） by bromate at pH 2.0. The reaction was monitored spectro- photometrically by measuring absorbance change with a fixed-time method of 5 min at 594 and 552 nm with and without surfactant. The variables influencing the calibra- tion sensitivity were extensively investigated, and the optimal conditions were established. The linear calibration range was 10-1,600 μg.L-1 with a relative SD ranging from 0.35 % to 3.35 % （for five replicate measurements of 75, 500, 1,000, and 1,500 μg.L-1） and a detection limit of 3.8 μg.L-1. The selectivity was also investigated, and greatly enhanced by suitable masking agents. The method was successfully applied to the analysis of V（IV） in pre- sence of excess V（V） up to 25 fold in environmental waters with the recoveries of 100.0 %-102.8 % for V（IV） and 95.7 %-99.7 % for total V. Its accuracy was validated by analysis of certified reference material via the present kinetic method and standard flame atomic absorption spectrometric method after extractive preconcentration with good agreement between certified and found values.

Immobilization and characterization of the Lactarius deliciosus mycelia-embedded polylysine-alginate beads and their decolorization performance

Liquid fermented fungal mycelia with decolorization capability have potential applications in scale-up. In this work, the Lactarius deliciosus mycelia were immobilized on ε-polylysine-alginate beads, and the decolorization effects of ε-polylysine-alginate beads were demonstrated along with Coomassie brilliant blue G-250 as a model dye. Morphology observation confirmed the beads had an exterior film and interior capsule with honeycomb microstructures suitable for mycelia growth. It was manifested that the maximum decolorization efficiency for mycelia was 98.5% at a removal rate of 0.68 mg·L^(-1)·h after 3 days. In comparison, the decolorization efficiency of the immobilized mycelia reached the maximum value of 97.3% at a removal rate of 6.1 mg·L^(-1)·h after 8 h. The enzyme activities of lignin peroxidase and laccase tested in the immobilized mycelia were significantly higher than in that of the free ones, such as the lignin peroxidase had the highest enzyme activity of 77.6 ± 7.4 U·L^(-1) in the former, while of 27.4 ± 8.7 U·L^(-1) in the latter. The immobilization of L. deliciosus mycelia could improve the decolorization of Coomassie brilliant blue G-250 efficiently. The prepared ε-polylysine-alginate beads embedded with L. deliciosus mycelia have very good reusability and a great potential in decolorizing analog dyes.