Biocatalytic production of enantiopure cyclohexane-trans-1,2-diol using extracellular lipases from Bacillus subtilis

Detry, Jean; Rosenbaum, Thorsten; Lütz, Stephan; Hahn, Doris; Jaeger, Karl‐Erich; Müller, Michael; Eggert, Thorsten

doi:10.1007/s00253-006-0391-9

Cited by 22 publications

(11 citation statements)

References 45 publications

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“…Product analysis revealed that only one diastereomer with anti 1,2-diacetoxy groups was prepared by the reaction of cyclohexene oxide with acetic anhydride, which gives exclusively the corresponding trans-1,2-diacetate. Stereochemical assignment was achieved by: i) comparison of the obtained 1 H NMR spectrum with one of authentic sample reported in the literature, 16 and ii) hydrolysis of rac-trans-1,2-diacetoxycyclohexane to white crystalline rac- …”

Section: Resultsmentioning

confidence: 99%

4Å molecular sieves catalyzed ring-opening of epoxides to 1,2-diacetates with acetic anhydride

Gilanizadeh¹,

Zeynizadeh²

2015

10.5267/j.ccl

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This study was aimed at highly stereoselective ring-opening of epoxides with acetic anhydride in the presence of molecular sieves 4Å as an efficient reagent. Yields of the 1,2-diacetates are uniformly good to excellent and the reagent may be recycled without the requirement for purification. High yields, short reaction times, ease of preparation and simple work-up procedure (filtration) are advantages of this method.

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Section: Resultsmentioning

confidence: 99%

4Å molecular sieves catalyzed ring-opening of epoxides to 1,2-diacetates with acetic anhydride

Gilanizadeh¹,

Zeynizadeh²

2015

10.5267/j.ccl

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“…Lyophilized BSA was purchased from Sigma-Aldrich (Taufkirchen, Germany). Lipases A and B from B. subtilis (BSL-A and BSL-B) were produced and purified as described previously [53]. Briefly, for large-scale production of BSL-A and BSL-B, high cell density fed-batch cultivation using E. coli BL21(DE3) as a host, carrying pET19b derived expression plasmids were performed in an Infors fermenter (30 L) [53-55].…”

Section: Methodsmentioning

confidence: 99%

“…Lipases A and B from B. subtilis (BSL-A and BSL-B) were produced and purified as described previously [53]. Briefly, for large-scale production of BSL-A and BSL-B, high cell density fed-batch cultivation using E. coli BL21(DE3) as a host, carrying pET19b derived expression plasmids were performed in an Infors fermenter (30 L) [53-55]. After purification by immobilized-metal affinity chromatography (IMAC) [56] using a Ni-nitrilo-triacetic acid (NTA) superflow column (30 mL, QIAGEN, Hilden, Germany), imidazole was removed by gel filtration chromatography (G-25 column, Amersham Pharmacia Biotech) using glycine/NaOH-buffer (10 mM, pH 10).…”

Section: Methodsmentioning

confidence: 99%

Mechanism of acetaldehyde-induced deactivation of microbial lipases

Franken

Eggert²,

Jaeger

et al. 2011

BMC Biochem

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BackgroundMicrobial lipases represent the most important class of biocatalysts used for a wealth of applications in organic synthesis. An often applied reaction is the lipase-catalyzed transesterification of vinyl esters and alcohols resulting in the formation of acetaldehyde which is known to deactivate microbial lipases, presumably by structural changes caused by initial Schiff-base formation at solvent accessible lysine residues. Previous studies showed that several lipases were sensitive toward acetaldehyde deactivation whereas others were insensitive; however, a general explanation of the acetaldehyde-induced inactivation mechanism is missing.ResultsBased on five microbial lipases from Candida rugosa, Rhizopus oryzae, Pseudomonas fluorescens and Bacillus subtilis we demonstrate that the protonation state of lysine ε-amino groups is decisive for their sensitivity toward acetaldehyde. Analysis of the diverse modification products of Bacillus subtilis lipases in the presence of acetaldehyde revealed several stable products such as α,β-unsaturated polyenals, which result from base and/or amino acid catalyzed aldol condensation of acetaldehyde. Our studies indicate that these products induce the formation of stable Michael-adducts at solvent-accessible amino acids and thus lead to enzyme deactivation. Further, our results indicate Schiff-base formation with acetaldehyde to be involved in crosslinking of lipase molecules.ConclusionsDifferences in stability observed with various commercially available microbial lipases most probably result from different purification procedures carried out by the respective manufacturers. We observed that the pH of the buffer used prior to lyophilization of the enzyme sample is of utmost importance. The mechanism of acetaldehyde-induced deactivation of microbial lipases involves the generation of α,β-unsaturated polyenals from acetaldehyde which subsequently form stable Michael-adducts with the enzymes. Lyophilization of the enzymes from buffer at pH 6.0 can provide an easy and effective way to stabilize lipases toward inactivation by acetaldehyde.

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“…Lipase A from Bacillus subtilis is a well‐characterised enzyme. It is one of the smallest lipases known, with a size of 19 kDa, and shows good hydrolytic activity in medium‐length substrates (Dartois et al ., ; Lesuisse et al ., ), having been used for the production of fine chemicals (Detry et al ., ). Its crystal structure is known (Van Pouderoyen et al ., ), making it possible to elucidate the structural basis of its stability at different pH (Rajakumara et al ., ), and site saturation mutagenesis has been used to study the role of specific amino acids in the properties of the enzyme (Fulton et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Bread making with Saccharomyces cerevisiae CEN.PK113‐5D expressing lipase A from Bacillus subtilis: leavening characterisation and aroma enhancement

Paciello

Landi

Orilio

et al. 2015

Int J of Food Sci Tech

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Lipase A from Bacillus subtilis was expressed in the yeast Saccharomyces cerevisiae CEN.PK113-5D strain as a cell wall-immobilised enzyme. The recombinant yeast was used in bread making to test the effect of lipase A on the bread properties such as rheological and aromatic properties. The results were compared to the not transformed strain and the commercial baker's yeast. The recombinant strain resulted a good leavening agent comparable to the commercial baker's yeasts provided 1% sucrose was added to the dough. Whereas, during the leavening, the trend of the rheological properties (cohesivness, gumminess and rigidity) differed from the commercial and the nontransformed (NT) strains. Moreover, using of the recombinant strain enhanced total aroma which achieved in the baked product a value of 9 lg per g product vs the 4 and 5 lg per g product of the NT strain and the commercial baker's yeast, respectively. To our knowledge, this is the first time that a bacterial lipase is used in bread making.Bread making with a lipase-recombinant yeast L. Paciello et al.

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Biocatalytic production of enantiopure cyclohexane-trans-1,2-diol using extracellular lipases from Bacillus subtilis

Cited by 22 publications

References 45 publications

4Å molecular sieves catalyzed ring-opening of epoxides to 1,2-diacetates with acetic anhydride

4Å molecular sieves catalyzed ring-opening of epoxides to 1,2-diacetates with acetic anhydride

Mechanism of acetaldehyde-induced deactivation of microbial lipases

Bread making with Saccharomyces cerevisiae CEN.PK113‐5D expressing lipase A from Bacillus subtilis: leavening characterisation and aroma enhancement

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