Protein engineering and applications of Candida rugosa lipase isoforms

Akoh, Casimir C.; Lee, Guan Chiun; Shaw, J. F.

doi:10.1007/s11745-004-1258-7

Cited by 102 publications

(77 citation statements)

References 86 publications

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“…In addition, it possesses stereoselectivity and regioselectivity ability for the synthesis of pharmaceuticals, glycoderivatives and carbohydrate esters used in the manufacture of many foods and fragrances [Akoh et al, 2004]. Previously, several li-pase genes with high homology (between 60 and 70%) had been cloned from C. rugosa, namely lip1 to lip7 [Longhi et al, 1992;Lotti et al, 1993].…”

Section: Introductionmentioning

confidence: 99%

Optimized Growth Kinetics of Pichia pastoris and Recombinant Candida rugosa LIP1 Production by RSM

et al. 2006

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A predictive model for Pichia pastoris expression of highly active recombinant Candida rugosa LIP1 was developed by combining the Gompertz function and response surface methodology (RSM) to evaluate the effect of yeast extract concentration, glucose concentration, temperature, and pH on specific responses. Each of the responses (maximum population densities, specific growth rate (µmax), protein concentration, and minimum lag phase duration) was determined using the modified Gompertz function. RSM and 4-factor-5-level central composite rotatable design (CCRD) were adopted to evaluate the effects of growth parameters, such as temperature (21.6–38.4°C), glucose concentration (0.3–3.7%), yeast extract (0.16–1.84%), and pH (5.3–8.7) on the responses of P. pastoris growth kinetics.Based on ridge maximum analysis, the optimum population density conditions were: temperature 24.4°C, glucose concentration 2.0%, yeast extract 1.5%, and pH 7.6. The optimum specific growth rate conditions were: temperature 28.9°C, glucose concentration 2.0%, yeast extract 1.1%, and pH 6.9. The optimum protein concentration conditions were: temperature 24.2°C, glucose concentration 1.9%, yeast extract 1.5%, and pH 7.6. Based on ridge minimum analysis, the minimal lag phase conditions were: temperature 32.3°C, glucose concentration 2.1%, yeast extract 1.1%, and pH 5.4. For the predicted value, the maximum population density, specific growth rate, protein concentration, and minimum lag phase duration were 15.7 mg/ml, 3.4 h^–1, 0.78 mg/ml, and 4.2 h, and the actual values were 14.3 ± 3.5 mg/ml, 3.6 ± 0.6 h^–1, 0.72 ± 0.2 mg/ml, and 4.4 ± 1.6 h, respectively.

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Section: Introductionmentioning

confidence: 99%

Optimized Growth Kinetics of Pichia pastoris and Recombinant Candida rugosa LIP1 Production by RSM

et al. 2006

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“…Therefore, esterases in general are attractive tools for biocatalytic applications, such as chiral synthesis of pharmaceuticals or agrochemicals (17). Recently, several bacterial esterases have been subjected to laboratory evolution to obtain enzymes with improved properties, including substrate specificity or enantioselectivity (2,33,35). The primary aim of the present study was to combine the advantages of cellular surface display with the attractive features of a bacterial esterase (4).…”

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confidence: 99%

Esterase Autodisplay: Enzyme Engineering and Whole-Cell Activity Determination in Microplates with pH Sensors

Schultheiss

Weiss

Winterer

et al. 2008

Appl Environ Microbiol

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Among the GDSL family of serine esterases/lipases is a group of bacterial enzymes that posses C-terminal extensions involved in outer membrane anchoring or translocation. ApeE from Salmonella enterica serovar Typhimurium, a member of this group, has been expressed in Escherichia coli and was resistant to protease digestion when the protease was added to whole cells, indicating a periplasmic localization. The five consensus blocks conserved within all GDSL esterases were identified in ApeE by multiple sequence alignment and separated from the C-terminal extension. The DNA sequence spanning the four invariant residues Ser, Gly, Asn, and His, and hence representing the catalytic domains of ApeE, was amplified by PCR and fused in frame to the transport domains of the autodisplay system. The resulting artificial esterase, called EsjA, was overexpressed in the cell envelope of E. coli and was shown to be active by the use of ␣-naphthyl acetate (␣-NA) as a substrate in an in-gel activity stain after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Surface exposure of EsjA was indicated by its accessibility to protease added to whole cells. The esterase activity of whole cells displaying EsjA was determined by a pH agar assay and by the use of microplates with integrated pH-dependent optical sensors. ␣-NA, ␣-naphthyl butyrate, and ␣-naphthyl caproate were used as substrates, and it turned out that the substrate preferences of artificial EsjA were altered in comparison to original ApeE. Our results indicate that autodisplay of esterase in combination with pH sensor microplates can provide a new platform technology for the screening of tailor-made hydrolase activities.

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“…The commercial lipase isoform mixtures prepared from Candida rugosa meet this requirement. Obtaining pure and different C. rugosa lipase isoforms is possible by means of computer modelling of lipase isoforms, and protein engineering methods such as lid swapping and DNA shuffling (Akoh et al, 2004). A recent review on microbial lipases focused on non-aqueous microbial lipase catalysis and major factors affecting esterification/transesterification processes in organic media.…”

Section: Application Name Example Reference(s)mentioning

confidence: 99%

“…Screening organic solvent-tolerant bacteria or extremophiles has been preferred to isolate and improve naturally solvent-stable enzymes (Gupta & Khare, 2009;Doukyu & Ogino, 2010). Other protein engineering examples with industrially and/or pharmacologically important enzymes include studies on cholesterol oxidase (Pollegioni et al, 2009), cyclodextrin glucanotransferases (Leemhuis et al, 2010), human butyrylcholinesterase (Masson et al, 2009), microbial glucoamylases (Kumar & Satyanarayana, 2009), lipases of different origins (Akoh et al, 2004;Verma et al, 2008;Kurtovic et al, 2009), phospholipases (Song et al, 2005;De Maria et al, 2007;Simockova & Griac, 2009) and phytases (Rao et al, 2009). Studies on extremozymes, enzymes isolated from extremophilic species, revealed their different structural and functional characteristics which could be exploited for biotechnological applications and improved further by protein engineering (Bjarnason et al, 1993;Hough & Danson, 1999;Georlette et al, 2004).…”

Section: Applications With Various Industrially Important Enzymesmentioning

confidence: 99%