Rational design of a new one-step purification strategy for Candida antarctica lipase B by ion-exchange chromatography

Trodler, Peter; Nieveler, Jens; Rusnak, Monika; Schmid, Rolf D.; Pleiss, Jürgen

doi:10.1016/j.chroma.2007.11.108

Cited by 42 publications

(29 citation statements)

References 20 publications

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“…The conventional purification process is usually begun by the removal of cells from the culture broth, either by centrifugation or by filtration. Then, the cell-free fermented broth is concentrated by ultrafiltration and ammonium sulphate precipitation, and isolated by chromatography and dialysis [8,9]. Liquid-liquid extraction using an aqueous two-phase system (ATPS) is an emerging and powerful technique for the purification of enzymes.…”

Section: Introductionmentioning

confidence: 99%

Purification of lipase produced by a new source of Bacillus in submerged fermentation using an aqueous two-phase system

Barbosa

Souza

Fricks

et al. 2011

Journal of Chromatography B

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Section: Introductionmentioning

confidence: 99%

Purification of lipase produced by a new source of Bacillus in submerged fermentation using an aqueous two-phase system

Barbosa

Souza

Fricks

et al. 2011

Journal of Chromatography B

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“…Cation exchange chromatography using an XK 16/20 column (Amersham Bioscience) filled with 7 mL source 15S resin was used for purification, according to the method of Trodler et al [10]. A SDS-PAGE image of the representative purified enzymes is shown in Fig.…”

Section: Calb Expression and Purificationmentioning

confidence: 99%

Simultaneous improvements in the activity and stability of Candida antarctica lipase B through multiple-site mutagenesis

Yagonia

Park

Hong

et al. 2015

Biotechnol Bioproc E

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An enzyme with improved characteristics is required for biochemical processes to be economically feasible. In this study, improvements in both the stability and activity of Candida antarctica lipase B (CALB) were integrated through multiple-site mutagenesis. CALB was divided into two regions to optimize its performance. Modulating the flexibility within the substrate-binding region and the hydrophilic solvent-affecting region can enhance the catalytic activity and organic solvent stability of CALB, respectively. Combining the mutation sites from the substrate-binding region and from the hydrophilic solvent-affecting region yielded an enzyme (V139E,A92E) with improved functionality. These findings suggest that the characteristics of CALB can be augmented by modulating its flexibility, and this method can possibly be applied to other lipases.

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“…7 Pseudozyma (Candida) antarctica is one of the most important sources of lipases for industrial applications. 8,9 Lipases from P. antarctica have many excellent characteristics and they are widely used for preparative purposes in organic synthesis in many industrial applications and scientific research projects. [10][11][12][13] However, the aim of research dealing with enzymes in non-aqueous media is to determine the optimal conditions for a specific application.…”

Section: Introductionmentioning

confidence: 99%

Production of lipase from Pseudozyma aphidis and determination of the activity and stability of the crude lipase preparation in polar organic solvents

Dimitrijević

Veličković

Bezbradica³

et al. 2011

JSCS

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The production of lipase from Pseudozyma aphidis (DSM 70725) was determined in six different media. The highest lipase production was observed in a medium with glucose as the sole carbon source, and yeast extract and sodium nitrate as the nitrogen sources. The time course studies of growth and lipase production in the optimal medium revealed that the highest lipase production was achieved at the end of the log phase of growth, reaching the value of 35.0 U cm -3 in the fifth day of cultivation. The effects of various polar, water-miscible, organic solvents on the activity and stability of the crude lipase produced by P. aphidis were evaluated. The hydrolytic activity of the crude lipase towards p-nitrophenyl palmitate (p-NPP) in aqueous media and in organic solvents was determined, using the same spectrophotometric assay in both the aqueous and organic media. The crude lipase preparation exhibited activity towards p-NPP only in acetone and acetonitrile, while the lipase was stable only in acetone, with 23 % residual activity after 24 h of incubation. These results suggested that lipase from P. aphidis can be used as a biocatalyst for potential applications in such organic solvents.

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Rational design of a new one-step purification strategy for Candida antarctica lipase B by ion-exchange chromatography

Cited by 42 publications

References 20 publications

Purification of lipase produced by a new source of Bacillus in submerged fermentation using an aqueous two-phase system

Purification of lipase produced by a new source of Bacillus in submerged fermentation using an aqueous two-phase system

Simultaneous improvements in the activity and stability of Candida antarctica lipase B through multiple-site mutagenesis

Production of lipase from Pseudozyma aphidis and determination of the activity and stability of the crude lipase preparation in polar organic solvents

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