Shiika Hara 1,2, Ayaka Koga 1,3, Yoshie Nagai-Yoshioka and ...
GluB20-2 protein with a 1,3-β-glucanase activity synthesis in bioreactor scale
1
New Biotechnology · Volume 29S · September 2012 have not been used for the detailed characterization of scalable bioconversion systems involving suspensions of immobilized bio- catalysts, and the effect of different operational conditions has not been assessed. Further work is being currently performed, to fur- ther establish the reproducibility of data throughout scales and biocatalyst stability. http://dx.doi.org/10.1016/j.nbt.2012.08.219 Poster 1.4.16 An NAPDH-dependent oxidoreductase from Saccha- romyces carlsbergensis as key enzyme in the synthesis of -lactones Enriqueta Martinez-Rojas 1 , Leif-Alexander Garbe 2 1 Research and Teaching Institute for Brewing in Berlin (VLB), Berlin, Germany 2 Technical University of Berlin, Berlin, Germany ,-Unsaturated carbonyl compounds are industrially impor- tant substances used as starting materials of several products, including plastics resins, dyes and pharmaceuticals. Because of the increasing demand for natural products, biotechnological processes for the production of lactones have been devel- oped. Generally, these processes involve the bioconversion of a hydroxy fatty acid to - or -lactones. In 2004 Garbe et al. have postulated the formation pathway of 4R--dodecalactone from 9S,10S-dihydroxyoctadecanoic acid in Saccharomyces cere- visiae, with 4-Oxo-2E-dodecenoic acid as intermediate. The latter was reduced by an enone reductase into 4-oxo-dodecanoic acid, further to 4R-hydroxy-dodecanoic acid and finally cycled to 4R--dodecanolactone. This study explored the biochemical per- formance of a purified enone reductase from Saccharomyces carlsbergensis and its role in the synthesis of enatiopure lac- tone. The enzyme was isolated and purified to homogeneity from 250 M 1-octen-3-one grown S. carlsbergensis. The NADPH- dependent oxidoreductase has a high activity toward different ,-unsaturated carbonyl compound. In addition, unsaturated alcohols are transformed into saturated ketones and alcohols. 13S-Hydroxy-9Z,11E-octadecadienoate was transformed into 13- oxo-9Z-octadeceoate and subsequently into -decalactone by yeast cells. (±)-4-Hydroxy-2E-decenoic acid ethyl ester was finally reduced to -decalactone. The result of this study have laid foun- dation for a more thorough understanding of how the yeast cells cope with oxidative stress, this has implication for industrial fields, since ,-unsaturated carbonyl compounds play a major role as raw material for synthesis of fine chemicals. http://dx.doi.org/10.1016/j.nbt.2012.08.220 Poster 1.4.17 GluB20-2 protein with a 1,3--glucanase activity synthe- sis in bioreactor scale Halina Zasłona ∗ , Anna Trusek-Hołownia Division of Chemical and Biochemical Processes, Faculty of Chemistry, Wroclaw University of Technology, Wybrze˙ ze St. Wyspia ´ nskiego 27, 50- 370 Wrocław, Poland In the XXI century, when the progressing deficit of non-renewable resources has become a problem on the world scale, the greater reliance began to put in the alternative source of carbon and energy – specially in the lignocellulosic materials. Lignocellulose is mainly plant biomass which is easy to receive. For that reason this kind of resources seems to be a very attractive substrate for bioconversion and that is why it has still more and more followers. The most often used enzyme in enzymatic hydrolysis of the lignocellulosic materials is mixture of cellulase and hemicellulase. Cellulase is usually mixture of egzo--1,4-glucanase, endo--1,4- glucanase and -glucanase. Hemicellulase is a mixture of different enzymes selected in dependence of lignocellulosic materials, which can be consist of diverse hemicelluloses. Some interesting specificities was identified in Escherichia coli genetically modified. In assigned condition, E. coli produces GluB20-2 – protein with a 1,3--glucanase activity. It is used to hydrolyze bonds in -glucan. This protein was produced in the scale bioreactors. The influ- ence of oxygen concentration, the intensity of mixing, the initial concentration of carbon and nitrogen sources on the amount of enzyme synthesized was tested. An increase in the intensity of biocatalyst production was observed in semi-continuous system. A method for initial purification of the enzyme in a semi-industrial scale was elaborated. The resulting preparation can be directly used in the reactions of hydrolysis of the lignocellulosic materials. http://dx.doi.org/10.1016/j.nbt.2012.08.221 Poster 1.4.18 Engineering of Bacillus subtilis physiological functional- ities towards the production of mycosubtilin anteiso-C17 N.E. Chihib, J. Guy, F. Coucheney, J.S. Guez, F. Coutte, M. Béchet, P. Jacques ∗ ProBioGEM, Laboratoire de Procédés Biologiques, Génie Enzymatique et Microbien, UPRES EA 1026, Polytech’Lille, IUT A, Université Lille 1 Sciences et Technologies, F-59655 Villeneuve d’Ascq Cedex, France The engineering of bacterial physiological functionalities can be realized by different ways such as strain adaptation and genetic modification. The main physiological characteristics for indus- trial applications are related to microbial metabolic capability, insensitivity of pathway key enzymes to end-product inhibition or feedback repression, robustness under adverse environmen- tal perturbations, tolerance of high concentration substrates or metabolites, and fitness throughout the entire production pro- cesses [1]. Mycosubtilin is synthesized by a hybrid polyketide syn- thase/nonribosomal synthetase by some strains of Bacillus subtilis. In the present work, our investigations aimed to find out selective www.elsevier.com/locate/nbt S79
Transcript of GluB20-2 protein with a 1,3-β-glucanase activity synthesis in bioreactor scale
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ew Biotechnology · Volume 29S · September 2012
ave not been used for the detailed characterization of scalableioconversion systems involving suspensions of immobilized bio-atalysts, and the effect of different operational conditions has noteen assessed. Further work is being currently performed, to fur-her establish the reproducibility of data throughout scales andiocatalyst stability.
ttp://dx.doi.org/10.1016/j.nbt.2012.08.219
oster 1.4.16
n NAPDH-dependent oxidoreductase from Saccha-omyces carlsbergensis as key enzyme in the synthesis of-lactones
nriqueta Martinez-Rojas1, Leif-Alexander Garbe2
Research and Teaching Institute for Brewing in Berlin (VLB), Berlin,ermanyTechnical University of Berlin, Berlin, Germany
�,�-Unsaturated carbonyl compounds are industrially impor-ant substances used as starting materials of several products,ncluding plastics resins, dyes and pharmaceuticals. Because ofhe increasing demand for natural products, biotechnologicalrocesses for the production of lactones have been devel-ped. Generally, these processes involve the bioconversion ofhydroxy fatty acid to �- or �-lactones. In 2004 Garbe et al.
ave postulated the formation pathway of 4R-�-dodecalactonerom 9S,10S-dihydroxyoctadecanoic acid in Saccharomyces cere-isiae, with 4-Oxo-2E-dodecenoic acid as intermediate. The latteras reduced by an enone reductase into 4-oxo-dodecanoic acid,
urther to 4R-hydroxy-dodecanoic acid and finally cycled toR-�-dodecanolactone. This study explored the biochemical per-ormance of a purified enone reductase from Saccharomycesarlsbergensis and its role in the synthesis of enatiopure lac-one. The enzyme was isolated and purified to homogeneityrom 250 �M 1-octen-3-one grown S. carlsbergensis. The NADPH-ependent oxidoreductase has a high activity toward different,�-unsaturated carbonyl compound. In addition, unsaturatedlcohols are transformed into saturated ketones and alcohols.3S-Hydroxy-9Z,11E-octadecadienoate was transformed into 13-xo-9Z-octadeceoate and subsequently into �-decalactone byeast cells. (±)-4-Hydroxy-2E-decenoic acid ethyl ester was finallyeduced to �-decalactone. The result of this study have laid foun-ation for a more thorough understanding of how the yeast cellsope with oxidative stress, this has implication for industrial fields,
ince �,�-unsaturated carbonyl compounds play a major role asaw material for synthesis of fine chemicals.ttp://dx.doi.org/10.1016/j.nbt.2012.08.220
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oster 1.4.17
luB20-2 protein with a 1,3-�-glucanase activity synthe-is in bioreactor scale
alina Zasłona ∗, Anna Trusek-Hołownia
Division of Chemical and Biochemical Processes, Faculty of Chemistry,roclaw University of Technology, Wybrzeze St. Wyspianskiego 27, 50-
70 Wrocław, Poland
n the XXI century, when the progressing deficit of non-renewableesources has become a problem on the world scale, the greatereliance began to put in the alternative source of carbon and energyspecially in the lignocellulosic materials. Lignocellulose is mainlylant biomass which is easy to receive. For that reason this kind ofesources seems to be a very attractive substrate for bioconversionnd that is why it has still more and more followers.
The most often used enzyme in enzymatic hydrolysis of theignocellulosic materials is mixture of cellulase and hemicellulase.ellulase is usually mixture of egzo-�-1,4-glucanase, endo-�-1,4-lucanase and �-glucanase. Hemicellulase is a mixture of differentnzymes selected in dependence of lignocellulosic materials,hich can be consist of diverse hemicelluloses. Some interesting
pecificities was identified in Escherichia coli genetically modified.n assigned condition, E. coli produces GluB20-2 – protein with a,3-�-glucanase activity. It is used to hydrolyze bonds in �-glucan.
This protein was produced in the scale bioreactors. The influ-nce of oxygen concentration, the intensity of mixing, the initialoncentration of carbon and nitrogen sources on the amount ofnzyme synthesized was tested. An increase in the intensity ofiocatalyst production was observed in semi-continuous system.method for initial purification of the enzyme in a semi-industrial
cale was elaborated. The resulting preparation can be directly usedn the reactions of hydrolysis of the lignocellulosic materials.
ttp://dx.doi.org/10.1016/j.nbt.2012.08.221
oster 1.4.18
ngineering of Bacillus subtilis physiological functional-ties towards the production of mycosubtilin anteiso-C17
.E. Chihib, J. Guy, F. Coucheney, J.S. Guez, F. Coutte, M. Béchet,. Jacques ∗
ProBioGEM, Laboratoire de Procédés Biologiques, Génie Enzymatiquet Microbien, UPRES EA 1026, Polytech’Lille, IUT A, Université Lille 1ciences et Technologies, F-59655 Villeneuve d’Ascq Cedex, France
he engineering of bacterial physiological functionalities can beealized by different ways such as strain adaptation and geneticodification. The main physiological characteristics for indus-
rial applications are related to microbial metabolic capability,nsensitivity of pathway key enzymes to end-product inhibitionr feedback repression, robustness under adverse environmen-al perturbations, tolerance of high concentration substrates or
etabolites, and fitness throughout the entire production pro-
esses [1].Mycosubtilin is synthesized by a hybrid polyketide syn-hase/nonribosomal synthetase by some strains of Bacillus subtilis.n the present work, our investigations aimed to find out selective
www.elsevier.com/locate/nbt S79
