Effects of organic solvents on activity and conformation of recombinant Candida antarctica lipase A produced by Pichia pastoris

Yang, Chunfang; Wang, Fanghua; Lan, Dongming; Whiteley, Chris G.; Yang, Bo; Wang, Yonghua

doi:10.1016/j.procbio.2011.11.017

Cited by 32 publications

(28 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Furthermore, solvents affect the catalytic proprieties, such as the selectivity lipases, and increase the substrate solubility, eliminating mass-transfer barriers for substrates [56]. Hence, the stability and activity of lipases in solvents are considered an important investigation point.…”

Section: Resultsmentioning

confidence: 99%

Lipolytic Potential ofAspergillus japonicusLAB01: Production, Partial Purification, and Characterisation of an Extracellular Lipase

Souza

Oliveira

Santos

et al. 2014

BioMed Research International

View full text Add to dashboard Cite

Lipolytic potential of Aspergillus japonicus LAB01 was investigated by describing the catalytic properties and stability of a secreted extracellular lipase. Enzyme production was considered high under room temperature after 4 days using sunflower oil and a combination of casein with sodium nitrate. Lipase was partially purified by 3.9-fold, resulting in a 44.2% yield using ammonium sulphate precipitation (60%) quantified with Superose 12 HR gel filtration chromatography. The activity of the enzyme was maximised at pH 8.5, and the enzyme demonstrated stability under alkaline conditions. The optimum temperature was found to be 45°C, and the enzyme was stable for up to 100 minutes, with more than 80% of initial activity remaining after incubation at this temperature. Partially purified enzyme showed reasonable stability with triton X-100 and was activated in the presence of organic solvents (toluene, hexane, and methanol). Among the tested ions, only Cu2+, Ni2+, and Al3+ showed inhibitory effects. Substrate specificity of the lipase was higher for C14 among various p-nitrophenyl esters assayed. The KM and V max values of the purified enzyme for p-nitrophenyl palmitate were 0.13 mM and 12.58 umol/(L·min), respectively. These features render a novel biocatalyst for industrial applications.

show abstract

Section: Resultsmentioning

confidence: 99%

Lipolytic Potential ofAspergillus japonicusLAB01: Production, Partial Purification, and Characterisation of an Extracellular Lipase

Souza

Oliveira

Santos

et al. 2014

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…The screenings were performed with a 5-fold molar excess of vinyl acetate as an irreversible acyl donor at ambient temperature. The selection of a proper solvent is a significant factor in biocatalysis [47–48], since solvents are able to change the activity [49–51] and enantioselectivity [52–54] of an enzyme. Conformational changes in the enzyme structure under the influence of solvent can even invert the substrate specificity and the enzyme–substrate affinity [55].…”

Section: Resultsmentioning

confidence: 99%

Chemoenzymatic synthesis and biological evaluation of enantiomerically enriched 1-(β-hydroxypropyl)imidazolium- and triazolium-based ionic liquids

Borowiecki¹,

Milner-Krawczyk²,

Plenkiewicz³

2013

Beilstein J. Org. Chem.

View full text Add to dashboard Cite

SummaryRacemic 1-(β-hydroxypropyl)azoles were prepared by solvent-free direct regioselective ring opening of 1,2-propylene oxide with imidazole or 1,2,4-triazole. Lipase-catalyzed transesterification of alcohols with vinyl acetate resulted in kinetic enantiomers resolution. Separated (S)-enantiomers of (+)-1-(1H-imidazol-1-yl)propan-2-ol and (+)-1-(1H-1,2,4-triazol-1-yl)propan-2-ol were quaternized with alkyl bromides or iodides, yielding novel optically active ionic liquids. Racemic salts were tested against a wide range of microorganisms.

show abstract

“…However,s ome lipases,s uch as lipase Af rom Candida antarctica (CAL-A) [3] and lipase 2f rom Candidap arapsilosis, [4] exhibit acyltransferase activity even in aqueous media( Scheme1). [6] The drawback of wild-type CAL-A is that in aqueous medium it is not totally selective towards transesterification:i ta lso acts as hydrolase, thereby releasing free fatty acids as as ide product, hence preventing its industrial use. In order to be competitive with chemical synthesis, the biocatalyst has to be robust in terms of temperature and solvents tability.…”

Section: Introductionmentioning

confidence: 99%

“…[5] CAL-Ai s generally considered asav ery stable enzyme and is therefore an interesting candidate for industrial ester synthesis. [6] The drawback of wild-type CAL-A is that in aqueous medium it is not totally selective towards transesterification:i ta lso acts as hydrolase, thereby releasing free fatty acids as as ide product, hence preventing its industrial use. [7] Protein engineering strategies, such as rational design,s emirational approaches, and random mutagenesis,a re able to modify lipases.…”

Section: Introductionmentioning

confidence: 99%

Enhancing the Acyltransferase Activity of Candida antarctica Lipase A by Rational Design

et al. 2015

View full text Add to dashboard Cite

A few lipases, such as Candida antarctica lipase A (CAL-A), are known to possess acyltransferase activity. This enables the enzyme to synthesize fatty acid esters from natural oils and alcohols even in the presence of bulk water. Unfortunately, fatty acids are still formed in these reactions as undesired side-products. To reduce the amount of fatty acids, several CAL-A variants were rationally designed based on its crystal structure. These variants were expressed in Escherichia coli and Pichia pastoris, purified, and their acyltransferase/hydrolase activities were investigated by various biocatalytic approaches. Among the investigated variants, mutant Asp122Leu showed a significant decrease in the hydrolytic activity, thus reducing the side-product yield during acylation. As desired, this variant retained wild-type process-relevant features like pH profile and thermostability.

show abstract

Effects of organic solvents on activity and conformation of recombinant Candida antarctica lipase A produced by Pichia pastoris

Cited by 32 publications

References 21 publications

Lipolytic Potential ofAspergillus japonicusLAB01: Production, Partial Purification, and Characterisation of an Extracellular Lipase

Lipolytic Potential ofAspergillus japonicusLAB01: Production, Partial Purification, and Characterisation of an Extracellular Lipase

Chemoenzymatic synthesis and biological evaluation of enantiomerically enriched 1-(β-hydroxypropyl)imidazolium- and triazolium-based ionic liquids

Enhancing the Acyltransferase Activity of Candida antarctica Lipase A by Rational Design

Contact Info

Product

Resources

About