1.

Yeast. 2010 Dec;27(12):1029-38. doi: 10.1002/yea.1812. Epub 2010 Sep 8.

Differences in hydrolytic abilities of two crude lipases from

Geotrichum candidum 4013.
Brabcov J, Zarevcka M, Mackov M

Source

Institute of Organic Chemistry and Biochemistry AS CR, Flemingovo nm. 2, 166 10 Prague 6,
Czech Republic.

Abstract

The fungus Geotrichum candidum 4013 produces two types of lipases (extracellular and cell-
bound). Both enzymes were tested for their hydrolytic ability to p-nitrophenyl esters and
compounds having a structure similar to the original substrate (triacylglycerols). Higher lipolytic
activity of extracellular lipase was observed when triacylglycerols of medium- (C12) and long-
(C18) chain fatty acids were used as substrates. Cell-bound lipase preferentially hydrolysed
trimyristate (C14). The differences in the abilities of these two enzymes to hydrolyse p-
nitrophenyl esters were observed as well. The order of extracellular lipase hydrolysis relation
velocity was as follows: p-nitrophenyl decanoate > p-nitrophenyl caprylate > p-nitrophenyl
laurate > p-nitrophenyl palmitate > p-nitrophenyl stearate. The cell-bound lipase indicates
preference for p-nitrophenyl palmitate. The most striking differences in the ratios between the
activity of both lipases (extracellular : cell-bound) towards different fatty acid methyl esters were
2.2 towards methyl hexanoate and 0.46 towards methyl stearate (C18). The Michaelis constant
(K(m) ) and maximum reaction rate (V(max) ) for p-nitrophenyl palmitate hydrolysis of cell-bound
lipase were significantly higher (K(m) 2.462 mM and V(max) 0.210 U/g/min) than those of
extracellular lipase (K(m) 0.406 mM and V(max) 0.006 U/g/min).

Copyright 2010 John Wiley & Sons, Ltd.

PMID:
20824885
[PubMed - indexed for MEDLINE]
Related citations

Publication Types, MeSH Terms, Substances

Publication Types
Comparative Study

MeSH Terms
Fungal Proteins/chemistry*
Fungal Proteins/genetics
Fungal Proteins/metabolism*
Geotrichum/chemistry
Geotrichum/enzymology*
Geotrichum/genetics
Hydrolysis
Kinetics
Lipase/chemistry*
Lipase/genetics
Lipase/metabolism*
 Substrate Specificity
Triglycerides/chemistry
Triglycerides/metabolism

Substances
Fungal Proteins
Triglycerides
Lipase

2.
Appl Microbiol Biotechnol. 2008 Mar;78(3):431-9. Epub 2008 Jan 10.

Optimization for producing cell-bound lipase from

Geotrichum sp. and synthesis of methyl oleate in
microaqueous solvent.
Yan JY, Yan YJ.

Source

College of Life Science and Technology, Huazhong University of Science and

Technology, 1037 Luoyu Road, Wuhan, 430074, China. yjiny@126.com

Abstract

An integrated optimization strategy involving a combination of different designs was

employed to optimize producing conditions of cell-bound lipase (CBL) from Geotrichum
sp. Firstly, it was obtained by a single factorial design that the most suitable carbon
source was a mixture of olive oil and citric acid and the most suitable nitrogen source
was a mixture of corn steep liquor and NH4NO3. Then, the Plackett-Burman design
was used to evaluate the effects of 13 variables related to CBL production, and three
statistically significant variables namely, temperature, olive oil concentration, and
NH4NO3 concentration, were selected. Subsequently, the levels of the three variables
for maximum CBL production were determined by response surface analysis as follows:
1.64% (v/v) olive oil, 1.49% (w/v) NH4NO3, and temperature 33.00 degrees C. Such
optimization resulted in a high yield of CBL at 23.15 U/ml, an enhanced 4.45-fold
increase relative to the initial result (5.2 U/ml) in shake flasks. The dried CBL was used
to synthesize methyl oleate in microaqueous hexane, resulting in 94% conversion after
24 h, and showed reusability with 70% residual activity and 69% conversion after eight
cycles of batch operation, which indicating that CBL, as a novel and natural form of
immobilized enzyme, can be effectively applied in repeated synthesis of methyl oleate
in a microaqueous solvent.

PMID:
18193214
[PubMed - indexed for MEDLINE]
Related citations

Publication Types, MeSH Terms, Substances

Publication Types
Research Support, Non-U.S. Gov't
 MeSH Terms
Culture Media/chemistry*
Enzyme Stability
Enzymes, Immobilized/metabolism
Geotrichum/enzymology*
Geotrichum/metabolism
Industrial Microbiology*
Lipase/chemistry
Lipase/metabolism*
Oleic Acids/biosynthesis*
Solvents/metabolism*

Substances
Culture Media
Enzymes, Immobilized
Oleic Acids
Solvents
methyl oleate
Lipase

3.
Bioresour Technol. 2004 Jan;91(1):77-84.

Optimization of extracellular lipase production by

Geotrichum sp. using factorial design.
Burkert JF, Maugeri F, Rodrigues MI.

Source

Department of Food Engineering, Universidade Estadual de Campinas,

Unicamp-- SP, CP 6121, Cep 13081-970, Campinas, Brazil.

Abstract

Response surface methodology was employed to study the effects of carbon

source (soy oil, olive oil and glucose) and nitrogen source concentrations (corn
steep liquor and NH(4)NO(3)) on the lipase production by Geotrichum sp. The
experiment included a 2(4) central composite rotatable design (CCRD) and four
others 2(3) CCRD. According to the responses from the experimental designs,
the effects of each variable were calculated and the interactions between them
were determined. The response surface methodology was applied for the
optimization of the nutrient concentrations in the culture medium for the enzyme
production, at 30 degrees C. The optimum medium composition for lipase
production by Geotrichum sp. was ammonium nitrate 2.1-2.5%, corn steep
liquor 13-15% and soy oil 0.6% as carbon source, which lead to a lipase activity
of about 20 U/ml. Using olive oil as carbon source, the optimum composition
was ammonium nitrate 0.8-1%, corn steep liquor 13-15% and olive oil 0.6%,
leading to an activity of 17 U/ml.

1.
Eur J Biochem. 1995 Mar 15;228(3):863-9.
Expression and characterization of Geotrichum candidum
lipase I gene. Comparison of specificity profile with lipase II.
Bertolini MC, Schrag JD, Cygler M, Ziomek E, Thomas DY, Vernet T.

Source

Biotechnology Research Institute, National Research Council of Canada, Montral, Qubec.

Abstract

Despite tremendous progress in the elucidation of three-dimensional structures of lipases, the

molecular basis for their observed substrate preference is not well understood. In an effort to
correlate the lipase structure with its substrate preference and to clarify the contradicting reports
in the literature, we have compared the enzymic characteristics of two closely related
recombinant lipases from the fungus Geotrichum candidum. These enzymes were expressed in
the yeast Saccharomyces cerevisiae as fusions with an N-terminal poly(His) tag and were
purified in a single step by metal-affinity chromatography. Their specific activities against a
series of triacylglycerol substrates were compared using a titrimetric assay. The substrates
varied in fatty acyl chain length, number of double bonds and their position along the chain. G.
candidum lipases I and II (GCL I and GLC II) are markedly different with respect to their
substrate preferences. For unsaturated substrates having long fatty acyl chains (C18:2 cis-9,
cis-12 and C18:3 cis-9, cis-12, cis-15), GCL I showed higher specific activity than GCL II,
whereas GCL II showed higher specific activity against saturated substrates having short fatty
acid chains (C8, C10, C12 and C14). We have constructed a hybrid molecule containing the N-
terminal portion of GCL I (including the flap covering the active site) linked to the C-terminal
portion of GCL II. The hybrid molecule showed a substrate preference pattern identical to that of
GCL II. These results indicate that sequence variation within the N-terminal 194 amino acids of
G. candidum lipases do not contribute to the observed variation in efficiency by which the
lipases hydrolyze their substrates. Moreover, it also shows that the flap region in GCL is not
directly involved in substrate differentiation, even though this region is thought to be involved in
recognition of the interface and in the activation of the enzyme.

PMID:
7737187
[PubMed - indexed for MEDLINE]
Free full text
Related citations

Publication Types, MeSH Terms, Substances

Publication Types
Comparative Study
Research Support, Non-U.S. Gov't

MeSH Terms
Base Sequence
Cloning, Molecular
Escherichia coli/genetics
Geotrichum/enzymology*
Geotrichum/genetics
Isoenzymes/genetics*
Isoenzymes/isolation & purification
Isoenzymes/metabolism
Lipase/genetics*
 Lipase/isolation & purification
Lipase/metabolism
Molecular Sequence Data
Oligodeoxyribonucleotides
Recombinant Fusion Proteins/genetics
Recombinant Fusion Proteins/metabolism

Substances
Isoenzymes
Oligodeoxyribonucleotides
Recombinant Fusion Proteins
lipase II, Geotrichum candidum
Lipase

2.
Biochim Biophys Acta. 1992 Jan 3;1123(1):59-64.

Substrate specificities of lipases A and B from

Geotrichum candidum CMICC 335426.
Charton E, Macrae AR.

Source

Unilever Research, Colworth Laboratory, Sharnbrook, Bedford, UK.

Abstract

The mould Geotrichum candidum produces extracellular lipases with different substrate
specificities according to strains. We purified two lipases - termed lipase A and lipase B
- from Geotrichum candidum CMICC 335426. The specificity of the two lipases was
investigated using hydrolysis assays on emulsions of pure acylglycerols and a wide
range of fatty acid esters. Lipase B was very highly specific for hydrolysis of esters of
cis-delta 9-fatty acids. Lipase A did not show such strict specificity, because it
hydrolysed a wider variety of fatty acid esters, in particular those of palmitic acid and
isomers of oleic acid. We think that differences in specificity previously observed for
crude lipases from various strains of G. candidum can be explained by the presence of
different levels of specific (lipase B) and non-specific (lipase A) lipases. As lipases A and
B are structurally related proteins, a minor variation in structure may be responsible for
the differing specificities.

Results: 4
1.
Appl Biochem Biotechnol. 2011 Aug;164(7):969-78. Epub 2011 Feb 8.

Isolation, identification and optimization of a new extracellular

lipase producing strain of Rhizopus sp.
Kantak JB, Bagade AV, Mahajan SA, Pawar SP, Shouche YS, Prabhune AA.

Source
National Chemical Laboratory, Dr. Homi Bhabha Road, Pashan, Pune, Maharashtra 411008,
India.

Abstract

A lipolytic mesophilic fungus which produces lipase extracellularly was isolated from soil. Based
on ITS1-5.8S-ITS4 region sequences of ribosomal RNA, it was concluded that the isolate JK-1
belongs to genus Rhizopus and clades with Rhizopus oryzae. The present paper reports the
screening, isolation, identification, and optimization of fermentation conditions for the production
of lipase (EC 3.1.1.3). Culture conditions were optimized, and the highest lipase production was
observed in basal medium with corn steep liquor as nitrogen source and glucose as carbon
source. Maximum lipase production was observed at 72 h, which is about 870 U/ml.
Optimization of fermentation conditions resulted in 16-fold enhancement in enzyme production.

PMID:
21302143
[PubMed - indexed for MEDLINE]
Related citations

MeSH Terms, Substances

MeSH Terms
Bacterial Proteins*/biosynthesis
Bacterial Proteins*/secretion
Biotechnology/methods*
Carbon/metabolism
Culture Media
DNA, Bacterial/analysis
Fermentation
Glucose/metabolism
Hydrogen-Ion Concentration
Kinetics
Lipase*/biosynthesis
Lipase*/secretion
Nitrogen/metabolism
Phylogeny
Rhizopus/classification
Rhizopus/enzymology*
Rhizopus/genetics
Rhizopus/isolation & purification
Sequence Analysis, DNA
Soil Microbiology*
Temperature
Zea mays/metabolism

Substances
Bacterial Proteins
Culture Media
DNA, Bacterial
Glucose
Carbon
Nitrogen
Lipase


2.
Appl Microbiol Biotechnol. 2011 Mar;89(6):1947-62. Epub 2010 Nov 13.

Quantitative study of lipase secretion, extracellular

lipolysis, and lipid storage in the yeast Yarrowia lipolytica
grown in the presence of olive oil: analogies with lipolysis
in humans.
Najjar A, Robert S, Gurin C, Violet-Asther M, Carrire F.

Source

Enzymologie Interfaciale et Physiologie de la Lipolyse, UPR 9025, CNRS, Aix-Marseille

Universit, Marseille, France.

Abstract

Lipase secretion, extracellular lipolysis, and fatty acid uptake were quantified in the
yeast Yarrowia lipolytica grown in the presence of olive oil and/or glucose. Specific
lipase assays, Western blot analysis, and ELISA indicated that most of the lipase
activity measured in Y. lipolytica cultures resulted from the YLLIP2 lipase. Lipase
production was triggered by olive oil and, during the first hours of culture, most of the
lipase activity and YLLIP2 immunodetection remained associated with the yeast cells.
YLLIP2 was then released in the culture medium before it was totally degraded by
proteases. Olive oil triglycerides were largely degraded when the lipase was still
attached to the cell wall. The fate of lipolysis products in the culture medium and inside
the yeast cell, as well as lipid storage, was investigated simultaneously by quantitative
TLC-FID and GC analysis. The intracellular levels of free fatty acids (FFA) and
triglycerides increased transiently and were dependent on the carbon sources. A
maximum fat storage of 37.8% w/w of yeast dry mass was observed with olive oil alone.
A transient accumulation of saturated FFA was observed whereas intracellular
triglycerides became enriched in unsaturated fatty acids. So far, yeasts have been
mainly used for studying the intracellular synthesis, storage, and mobilization of neutral
lipids. The present study shows that yeasts are also interesting models for studying
extracellular lipolysis and fat uptake by the cell. The quantitative data obtained here
allow for the first time to establish interesting analogies with gastrointestinal and
vascular lipolysis in humans.

PMID:
21076918
[PubMed - indexed for MEDLINE]
Related citations

Publication Types, MeSH Terms, Substances

Publication Types
Research Support, Non-U.S. Gov't

MeSH Terms
Blotting, Western
Chromatography, Gas
Chromatography, Thin Layer
 Culture Media/chemistry
Cytosol/chemistry
Enzyme-Linked Immunosorbent Assay
Glucose/metabolism
Lipase/secretion*
Lipid Metabolism*
Plant Oils/metabolism*
Yarrowia/growth & development
Yarrowia/metabolism*

Substances
Culture Media
Plant Oils
Glucose
olive oil
Lipase

3.
Fungal Biol. 2010 Nov-Dec;114(11-12):911-6. Epub 2010 Sep 17.

The entomopathogen Metarhizium anisopliae can

modulate the secretion of lipolytic enzymes in
response to different substrates including
components of arthropod cuticle.
Beys da Silva WO, Santi L, Corra AP, Silva LA, Bresciani FR, Schrank A,
Vainstein MH.

Source

Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, P.O. Box

15005, CEP 91501-970, Porto Alegre, RS, Brazil. walterbeys@cbiot.ufrgs.br

Abstract

The filamentous fungus Metarhizium anisopliae is a well-characterized,

arthropod pathogen used in the biological control of arthropod pests. Studies on
the regulation of enzymes related to host infection such as proteases and
chitinases have been reported but little is known about regulation of lipolytic
enzymes in this fungus. Here we present the effects of different carbon sources
such as components of the arthropod cuticle on the secretion of lipolytic
enzymes by M. anisopliae. Differences in the induction of lipolytic activity were
observed between the several carbon sources tested. Higher activities of lipase
or lipase/esterase were found in culture media containing the arthropod
integument components chitin and cholesteryl stearate. Several bands of
lipolytic activity were also detected in zymograms, thus suggesting an important
set of lipolytic enzymes secreted by the fungus. These results show that the
fungus can modulate the secretion of lipolytic activity in response to host
integument components, thus reinforcing the potential role of these enzymes
during M. anisopliae infection.
 Copyright 2010 The British Mycological Society. Published by Elsevier Ltd. All
rights reserved.

PMID:
21036334
[PubMed - indexed for MEDLINE]
Related citations

Publication Types, MeSH Terms, Substances

Publication Types
Research Support, Non-U.S. Gov't

MeSH Terms
Animals
Arthropods/growth & development
Arthropods/microbiology*
Esterases/metabolism*
Fungal Proteins/metabolism*
Host-Pathogen Interactions*
Lipase/metabolism*
Metarhizium/enzymology*
Metarhizium/physiology
Protein Transport

Substances
Fungal Proteins
Esterases
Lipase

4.
Appl Microbiol Biotechnol. 2011 Jan;89(1):109-19. Epub 2010 Sep 7.

A molecular approach to optimize hIFN 2b

expression and secretion in Yarrowia lipolytica.
Gasmi N, Fudalej F, Kallel H, Nicaud JM.

Source

INRA, UMR1319 Micalis, Domaine de Vilvert, 78352 Jouy-en-Josas,

France.

Abstract

In this work, we investigated the effect of codon bias and consensus

sequence (CACA) at the translation initiation site on the expression
level of heterologous proteins in Yarrowia lipolytica; human interferon
alpha 2b (hIFN-2b) was studied as an example. A codon optimized
hIFN-2b gene was synthesized according to the frequency of codon
usage in Y. lipolytica. Both wild-type (IFN-wt) and optimized hIFN-2b
 (IFN-op) genes were expressed under the control of a strong inducible
promoter acyl-co-enzyme A oxidase (POX2). Protein secretion was
directed by the targeting sequence of the extracellular lipase (LIP2):
pre-proLIP2. Codon optimization increased protein production by 11-
fold, whereas the insertion of CACA sequence upstream of the initiation
codon of IFN-op construct resulted in 16.5-fold increase of the
expression level; this indicates that translational efficiency plays an
important part in the increase of hIFN-2b production level. The
replacement of the pre-proLIP2 signal secretion with the LIP2 pre-
region sequence followed by the X-Ala/X-Pro stretch but without the
pro-region also increased the secretion of the target protein by twofold,
suggesting therefore that the LIP2 pro-region is not necessary for
extracellular secretion of small heterologous proteins in Yarrowia
lipolytica.

6.
Lipids. 1995 Aug;30(8):747-54.

Substrate specificity of the lipase from Candida parapsilosis.

Briand D, Dubreucq E, Grimaud J, Galzy P.

Source

Ecole Nationale Suprieure Agronomique, Institut National de la Recherche Agronomique,

Montpellier, France.

Abstract

Substrate specificity of the acyltransferase activity of the lipase (EC 3.1.1.3) from Candida
parapsilosis CBS 604 was studied in aqueous media. The specificity toward both acid and
alcohol parts of a large number of acylglycerols and aliphatic esters was investigated. This
lipase showed a high activity in the presence of esters with long-chain fatty acids and
particularly unsaturated fatty acids with a cis-delta 9 double bond. It was observed that the
activity profile depended not only on the alcohol part of the acyl ester, but also on the
temperature of the reactant medium. The best lipid substrates had their melting point between
-40 to +20 degrees C, 14 to 18 carbon atoms in the acyl group and 1 to 4 carbon atoms in the
alkyl group. The enzyme, defined as an acyltransferase in a previous paper, showed a high
affinity for primary and secondary alcohols with a short carbon chain (1 to 5 carbon atoms) as
acyl acceptors. The influence of free alcohols in the reactant medium on the hydrolysis and
alcoholysis activities of the enzyme is discussed. Two phenomena seem to be involved,
depending on the alcohol: competition with water for the acyltransfer reaction and lipid substrate
dilution when the alcohol places at the oil/water interface.

10.
Appl Biochem Biotechnol. 2007 May-Jun;141(2-3):377-90.

Screening and optimization of media constituents for

enhancing lipolytic activity by a soil microorganism using
statistically designed experiments.
Haider MA, Pakshirajan K.

Source

Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam,

781039, India.
Abstract

Soil contaminated with vegetable cooking oil was used in the isolation of a lipase-producing
microorganism. The effectiveness of two different statistical design techniques in the screening
and optimization of media constituents for enhancing the lipolytic activity of the soil
microorganism was determined. The media constituents for lipase production by the isolated
soil microorganism were screened using a Plackett-Burman design. Oil, magnesium sulfate, and
ferrous sulfate were found to influence lipolytic activity at 24 and 72 h of culture with very high
confidence levels. Whereas oil and ferrous sulfate showed a positive effect, magnesium sulfate
indicated a negative effect on the lipolytic activity. A central composite design (CCD) followed by
response surface methodology was used in optimizing these media constituents for enhancing
the lipolytic activity. The regression model obtained for 72 h of lipolytic activity was found to be
the best fit, with R 2=0.97, compared with the other model. An optimum combination at 9.3 mL/L
of oil, 0.311 g/L of magnesium sulfate, and 0.007 g/L of ferrous sulfate in the media gave a
maximum measured lipolytic activity of 7.1 U/mL at 72 h of culture. This increase in lipolytic
activity was found to be 10.25% higher than the maximum experimentally observed value in the
CCD.

12.
Can J Microbiol. 2007 May;53(5):643-55.

Optimization of medium composition for lipase production by

Candida rugosa NCIM 3462 using response surface
methodology.
Rajendran A, Thangavelu V.

Source

Biochemical Engineering Laboratory, Department of Chemical Engineering, Annamalai

University, Annamalai Nagar - 608 002, Tamil Nadu, India. donaravind@yahoo.com

Abstract

A sequential optimization approach using statistical design of experiments was employed to

enhance the lipase production by Candida rugosa in submerged batch fermentation. Twelve
medium components were evaluated initially using the Plackett-Burman 2-level factorial design.
The significant variables affecting lipase production were found to be glucose, olive oil, peptone,
(NH4)2SO4, and FeCl3.6H2O. Various vegetable oils were tested in the second step, and
among them, groundnut oil was found to be the best inducer for lipase production by C. rugosa.
The third step was to identify the optimal values of the significant medium components with
groundnut oil as the inducer using response surface methodology. The regression equation
obtained from the experimental data designed using a central composite design was solved,
and analyzing the response surface contour plots, the optimal concentrations of the significant
variables were determined. A maximum lipase activity of 5.95 U.mL-1, which is 1.64 times the
maximum activity obtained in the Plackett-Burman experimental trials, was observed. The
optimum combination of medium constituents contained 19.604 g.L-1 glucose, 13.065 mL.L-1
groundnut oil, 7.473 g.L-1 peptone, 0.962 g.L-1 (NH4)2SO4, 0.0019 g.L-1 FeCl3.6H2O, and
other insignificant components at the fixed level. A predictive model of the combined effects of
the independent variables using response surface methodology and an artificial neural network
was proposed. The unstructured kinetic models, logistic model, and Luedeking-Piret model were
used to describe cell mass and lipase production. The parameters of the models were evaluated
and the lipase production by C. rugosa was found to be growth associated.

13.
J Mol Microbiol Biotechnol. 2006;11(1-2):28-40.
Optimized growth kinetics of Pichia pastoris and recombinant
Candida rugosa LIP1 production by RSM.
Chang SW, Shieh CJ, Lee GC, Akoh CC, Shaw JF.

Source

Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.

Abstract

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 (mumax), 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 degrees 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 degrees
C, glucose concentration 2.0%, yeast extract 1.5%, and pH 7.6. The optimum specific growth
rate conditions were: temperature 28.9 degrees C, glucose concentration 2.0%, yeast extract
1.1%, and pH 6.9. The optimum protein concentration conditions were: temperature 24.2
degrees 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 degrees 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.

17.
Biotechnol Bioeng. 1996 Aug 5;51(3):371-4.

Optimized enzymatic synthesis of geranyl butyrate with lipase

AY from Candida rugosa.
Shieh CJ, Akoh CC, Yee LN.

Source

Department of Food Science and Technology, Food Science Building, The University of
Georgia, Athens, Georgia 30602-7610, USA.

Abstract

Response surface methodology (RSM) and five-level, five-variable central composite rotatable
design (CCRD) were used to evaluate the effects of synthetic variables, such as reaction time
(1-9 h), temperature (25-65 degrees C), enzyme amount (10-50%), substrate molar ratio of
geraniol to tributyrin (1:0.33-1:1), and added water amount (0-20%) on molar percent yield of
geranyl butyrate, using lipase AY from Candida rugosa. Reaction time and temperature were the
most important variables and substrate molar ratio had no effect on percent molar conversion.
Based on contour plots, optimum conditions were: reaction time 9 h, temperature 35 degrees C,
enzyme amount 50%, substrate molar ratio 1:0.33, and added water 10%. The predicted value
was 100% and actual experimental value was 96.8% molar conversion.