Method for B-phycoerythrin purification from Porphyridium cruentum

Tchernov, A.A.; Minkova, Kaledona; Georgiev, D.; Houbavenska, N. B.

doi:10.1007/bf00156361

Cited by 17 publications

(10 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3). These spectra agree well with those published references for pure B-PE [9,11,12,30,36,37]. The homogeneity and structure integrity of the B-PE obtained from the SOURCE 15Q column were confirmed by the native PAGE which showed only one band (Fig.…”

Section: Source 15q Chromatography and Characterization Of B-pesupporting

confidence: 91%

“…Moreover, SOURCE 15Q can be applied on FPLC, which can be readily scaled to industrial production of kilograms of purified protein in columns with volumes of many liters. The conventional chromatography methods employed in the purification of B-PE from P. cruentum involve gel filtration (Sephadex), DEAE chromatography, expanded bed adsorption (EBA) chromatography, often in combination [11,12,18,30]. By our protocol, using only a one-step SOURCE 15Q chromatography method, purity ratio of the B-PE of P. cruentum was enhanced by a factor of 2.6.…”

Section: Comparison Of the Purification Efficiency Of Different Chrommentioning

confidence: 99%

See 1 more Smart Citation

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

Tang

Zhao

Bao

et al. 2016

Protein Expression and Purification

View full text Add to dashboard Cite

a b s t r a c t B-phycoerythrin (B-PE) was separated and purified from microalga Porphyridium cruentum using onestep chromatographic method. Phycobiliproteins in P. cruentum was extracted by osmotic shock and initially purified by ultrafiltration. Further purification was carried out with a SOURCE 15Q exchange column and analytical grade B-PE was obtained with a purity ratio (A 545 /A 280 ) of 5.1 and a yield of 68.5%. It showed a double absorption peaks at 545 nm and 565 nm and a shoulder peak at 498 nm, and displayed a fluorescence emission maximum at 580 nm. The analysis by sodium dodecyl sulfate epolyacrylamide gel electrophoresis (SDSePAGE) showed a bulky band between 18 and 20 kDa which could be assigned to subunits a and b and a low intensity band of 27 kDa assigned to g subunit. Our protocol provides attractive alternative to consider for the purification procedure to obtain analytical grade B-PE at commercial level.

show abstract

Section: Source 15q Chromatography and Characterization Of B-pesupporting

confidence: 91%

Section: Comparison Of the Purification Efficiency Of Different Chrommentioning

confidence: 99%

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

Tang

Zhao

Bao

et al. 2016

Protein Expression and Purification

View full text Add to dashboard Cite

show abstract

“…So we designed to use rivanol to absorb im-purities in 5 gradients' solutions in which rivanol to protein ratios (g/g) ranged from 1:10 to 16:10, and achieved a purity of 0.56 and 0.28 for PE and PC, respectively. These results were in contrast with previous reports showing that rivanol could remove polysaccharide sharply from solutions [30]. Our results indicated that precipitations with other reagents, e.g., active carbon and rivanol did not contribute to the purification of phycobiliproteins from P. yezoensis.…”

Section: Primary Isolation Of Pbp From P Yezoensiscontrasting

confidence: 99%

“…Rivanol was more widely utilized according to previous reports. It was applied to isolate PBP from Porphyridium cruentum [30] and Nostoc sp [31]. CPC with purity of 3.90 and 3.00 from Spirulina (Arthrospira) fusiformis [32] and Arthronema africanum [29], respectively, were purified after fractional precipitation with rivanol.…”

Section: Primary Isolation Of Pbp From P Yezoensismentioning

confidence: 99%

Large scale preparation of phycobiliproteins from <i>Porphyra yezoensis</i> using co-precipitation with ammonium sulfate

Cai¹,

Li²,

Wu³

et al. 2012

View full text Add to dashboard Cite

The purpose of this study was to extract phycobiliproteins (PBP) from Porphyra yezoensis using an optimized procedure and further establish a large scale process for protein production. According to our previous experiences on the extraction of PBP, salting out methods, e.g. ammonium sulfate precipitation, worked more efficiently than isoelectric precipitation, differential centrifugation or ultrafiltration. Thus, we chose ammonium sulfate to coprecipitate PBP in crude solution. After four times of precipitation followed by one time of high speed centrifugation, the maximum purity of crude phycoerythrin and phycocyanin reached 1.94 (A 565 /A 280 ) and 0.85 (A 615 /A 280 ), with a yield of 0.50% and 0.37%, respectively. A total of 0.94 mg phycoerythrin and 0.54 mg phycocyanin with purity of more than 3.2 were obtained from 1 g dried P. yezoensis after additional chromatography. We further scaled up the frozen dried P. yezoensis from 20 g to 400 g, with 1295 mg phycoerythrin and 593 mg phycocyanin obtained.

show abstract

“…The precipitation agent, 6,9-diamino-2-ethoxyacridine lactate (Ethodin), is a highly aromatic, cationic dye that complexes with negatively charged components, e.g., proteins, DNA, and cell debris (Franek, 1986;Miller, 1959;Neurath and Brunner, 1969;Tchernov et al, 1993) (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Purification of antibody and antibody‐fragment from E. coli homogenate using 6,9‐diamino‐2‐ethoxyacridine lactate as precipitation agent

Persson

Lester

2004

Biotech & Bioengineering

View full text Add to dashboard Cite

To obtain a more efficient purification process for antibody fragments from an Escherichia coli homogenate, the precipitant, Ethodin (6,9-diamino-2-ethoxyacridine lactate) was introduced to the homogenate. By adding the precipitant a drastic reduction of host cell protein was obtained. The majority of the proteins were recovered in a precipitate with the cell debris, while the antibody or antibody-fragment was recovered in the clarified supernatant. In addition, DNA was also efficiently precipitated when using Ethodin as a precipitation agent. The improved purity of the clarified extract obtained by using the precipitant allows for the use of smaller chromatography columns and may reduce the number of chromatographic steps required in the recovery process. The effect of Ethodin concentration, pH, temperature, and conductivity were investigated. The investigation was performed on two different antibody-fragments, e.g., F(abV) 2 molecules and a full-length antibody produced in E. coli. The two F(abV) 2 proteins were F(abV) 2 A and F(abV) 2 B, which have a similar molecular mass (100 kDa) but different isoelectric points (pIs), i.e., 8.9 and 7.5, respectively. The full-length antibody, Ab (the full IgG form of F(abV) 2 B) has a pI of 7.8 and molecular mass of 150 kDa. The investigation showed that the highest purification factors were obtained at neutral pH, low conductivity, and Ethodin concentrations of 0.6%. B

show abstract

Method for B-phycoerythrin purification from Porphyridium cruentum

Cited by 17 publications

References 15 publications

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

Large scale preparation of phycobiliproteins from <i>Porphyra yezoensis</i> using co-precipitation with ammonium sulfate

Purification of antibody and antibody‐fragment from E. coli homogenate using 6,9‐diamino‐2‐ethoxyacridine lactate as precipitation agent

Contact Info

Product

Resources

About

Method for B-phycoerythrin purification from Porphyridium cruentum

Cited by 17 publications

References 15 publications

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

One-step chromatographic procedure for purification of B-phycoerythrin from Porphyridium cruentum

Large scale preparation of phycobiliproteins from &lt;i&gt;Porphyra yezoensis&lt;/i&gt; using co-precipitation with ammonium sulfate

Purification of antibody and antibody‐fragment from E. coli homogenate using 6,9‐diamino‐2‐ethoxyacridine lactate as precipitation agent

Contact Info

Product

Resources

About

Large scale preparation of phycobiliproteins from <i>Porphyra yezoensis</i> using co-precipitation with ammonium sulfate