Purification of <i>Aspergillus carbonarius</i> polygalacturonase using polymeric membranes

Nakkeeran, Ekambaram; Venkatesh, Krishnappa; Subramanian, R.; Kumar, Sudhir

doi:10.1002/jctb.1895

Cited by 8 publications

(5 citation statements)

References 20 publications

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“…Earlier experiments using several UF membranes (10, 25, 100, and 500 kDa) showed that the 10 kDa membrane in diafiltration operation removed most of the carbohydrates, phosphate salts, and total acids from A. carbonarius culture broth with a higher PG recovery [11]. Although the protocol was effective in eliminating non-protein impurities, there was practically no improvement in the enzyme specific activity since contaminating proteins were also recovered along with the enzyme.…”

Section: Resultsmentioning

confidence: 85%

“…membranes ( Table 3). The first two steps were employed for pre-processing the crude culture broth to eliminate non-protein impurities as described earlier [11]. Processing the above culture broth after suitable dilution through 450 nm MF membrane eliminated 21% larger contaminating proteins.…”

Section: Purification Of Pg (Cf-uf-mf-uf)mentioning

confidence: 99%

“…In an earlier study [11], we reported near complete elimination of carbohydrates, salts, and acids from A. carbonarius culture broth containing PG with 96% recovery using a 10 kDa UF membrane. The protocol developed improved the enzyme activity from 4.6 to 49.4 U/mg of dry matter mainly by eliminating interfering carbohydrates that affect concentration processes such as lyophilization and precipitation [3].…”

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confidence: 96%

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Improving Specific Activity of Aspergillus carbonarius Polygalacturonase Using Polymeric Membranes

Nakkeeran

Subramanian

Kumar

2008

Appl Biochem Biotechnol

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Microfiltration (MF) and ultrafiltration (UF) membranes were screened for improving the specific activity of polygalacturonases (PG) in the culture broth of Aspergillus carbonarius obtained after submerged fermentation. While 200 and 450 nm MF membranes eliminated some of the larger non-enzymatic proteins, 50 kDa UF membrane exhibited a marginal selectivity between the enzyme and other smaller proteins. The 450 nm MF and 50 kDa UF membranes selected were further evaluated under different process conditions for an integrated membrane process. The process efficacy of three different schemes was also studied for enzyme purification. A two-stage membrane process employing MF followed by UF improved the enzyme-specific activity (5,590 U/mg) by 4.69-fold eliminating the larger and smaller non-enzymatic proteins as well as non-protein impurities with a recovery of 76% enzymes, besides resulting in higher productivity. Thus, adoption of integrated membrane process with appropriate selection of membranes could result in high recovery of enzymes with improved specific activity.

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Section: Resultsmentioning

confidence: 85%

Section: Purification Of Pg (Cf-uf-mf-uf)mentioning

confidence: 99%

mentioning

confidence: 96%

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Improving Specific Activity of Aspergillus carbonarius Polygalacturonase Using Polymeric Membranes

Nakkeeran

Subramanian

Kumar

2008

Appl Biochem Biotechnol

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“…However, in spite of widespread use of ultrafiltration in processes such as diafiltration and concentration, the potential for its use in protein fractionation has not been exploited in the biotechnology industry [17]. Nevertheless, there are studies that show that ultrafiltration can be applied to purification of enzymes, obtaining high yields and product purity at the same time [19][20][21].…”

Section: Introductionmentioning

confidence: 99%

One-Step Ultrafiltration Process for Separation and Purification of a Keratinolytic Protease Produced with Feather Meal

Machado

Severo

Oliveira

et al. 2018

International Journal of Chemical Engineering

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A purification technique to obtain keratinolytic proteases produced by Bacillus sp. P45 in a medium containing chicken feather meal as substrate is presented. e experiments were carried out in a dead-end ultrafiltration unit, and the influence of the membrane cutoff, pH of enzymatic extract, and operating pressure on the purification of keratinase were studied. e one-step ultrafiltration process with the membrane molecular mass cutoff of 10 kDa at pH 8.0 and operating pressure of 0.147 MPa showed an enzyme recovery of 87.8% and a 4.1-fold purification factor. It is showed that ultrafiltration could be potentially used in the purification of keratinases.

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“…Although a lot of recent studies have demonstrated the possibility of proteins fractionation by UF, some of them dealt with simulated binary or ternary mixtures [29,30]. Experimental studies of real biological streams are not very often presented [31][32][33][34][35].…”

Section: Introductionmentioning

confidence: 99%

Effective Purification of Cerrena unicolor Laccase Using Microfiltration, Ultrafiltration and Acetone Precipitation

Bryjak

Rekuć

2009

Appl Biochem Biotechnol

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Microfiltration followed by concentration and diafiltration on ultrafiltration membranes (Biomax-100, Biomax-10 and Ultracel-10) was used to recover extracellular laccase (EC 1.10.3.2) from culture broth of wood-rotting fungus Cerrena unicolor. Feed, permeate, retentate and membrane wash-out solutions were analysed for the presence of laccase, proteases, protein and brown impurities. An easy, cheap and short-term procedure was proposed to obtain retentates with low yields of total proteins (less than 14%), proteases activity (less than 15%) and brown impurities (from 2% to 29%) with a simultaneous laccase recovery above 73%. The degree of laccase purification varied from 6.7 to 11.0 and depended on the type of membrane used and content of brown pigments in the feed. Subsequent protein precipitation with cold acetone increased the degree of purification about twice and reduced proteases and brown impurities to some extent. Ultracel-10 membrane was recommended as the best solution to prevent fouling of membranes and to obtain laccase-enriched fraction with a very low content of proteases and brown pigments.

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Purification of Aspergillus carbonarius polygalacturonase using polymeric membranes

Cited by 8 publications

References 20 publications

Improving Specific Activity of Aspergillus carbonarius Polygalacturonase Using Polymeric Membranes

Improving Specific Activity of Aspergillus carbonarius Polygalacturonase Using Polymeric Membranes

One-Step Ultrafiltration Process for Separation and Purification of a Keratinolytic Protease Produced with Feather Meal

Effective Purification of Cerrena unicolor Laccase Using Microfiltration, Ultrafiltration and Acetone Precipitation

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