Preparation of Detergent-Lipase Complexes Utilizing Water-Soluble Amphiphiles in Single Aqueous Phase and Catalysis of Transesterifications in Homogeneous Organic Solvents.

Mine, Yurie; Fukunaga, Kimitoshi; MARUOKA, NOBUAKI; Nakao, Katsumi; Sugimura, Yoshiaki

doi:10.1263/jbb.90.631

Cited by 10 publications

(6 citation statements)

References 22 publications

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“…Detergents are able to increase the activity of free and immobilized lipases. [25][26][27][28] Detergents are suggested to be able to shift the open/close equilibrium of lipases toward the open conformation, by coating the hydrophobic areas of the lipases that surround the active center of the enzyme. This coating would reduce the negative interactions between this large and highly hydrophobic pocket and the environment.…”

Section: Discussionmentioning

confidence: 99%

Glutaraldehyde Cross-Linking of Lipases Adsorbed on Aminated Supports in the Presence of Detergents Leads to Improved Performance

et al. 2006

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Lipases from Candida rugosa (CRL) and lipase isoforms A and B from Candida antarctica (CAL-A and CAL-B) were adsorbed on aminated supports in the presence of detergents to have individual lipase molecules. Then, one fraction was washed to eliminate the detergent, and both preparations were treated with glutaraldehyde. The presence of detergent during the cross-linking of the lipases to the support permitted an increase in the recovered activity (in some instances, even by a 10-fold factor). This activity was higher even than that exhibited by the just adsorbed lipases, suggesting that it was not a result of some protective effect of the detergent in the enzyme activity during glutaraldehyde chemical modification. Moreover, the enantioselectivity of the different enzyme preparations was very different if the glutaraldehyde was offered in the presence or in the absence of detergent, in some cases increasing the E value (even by a 7-fold factor in the case of CAL-A in the hydrolysis of (+/-)-2-hydroxy-4-phenylbutyric acid ethyl ester), in other cases even inverting the enantio preference (e.g., in the case of CRL). The irreversible chemical inhibition of the enzyme that was immobilized and cross-linked with glutaraldehyde in the presence of detergents was more rapid than that in the other preparations (by more than a 10-fold factor). This experiment reveals an exposition degree of the active serine in the preparation cross-linked with the support in the presence of detergent that is higher than that in the other preparations. The results suggested that different enzyme structures were "stabilized" by the glutaraldehyde treatment if performed in the presence or in the absence of detergent, and that, in the presence of detergent, a form of the lipase with the serine residue more exposed to the medium and much more active could be obtained. This strategy seems to be of general use to improve the lipase activity to be used in macroaqueous media.

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Section: Discussionmentioning

confidence: 99%

Glutaraldehyde Cross-Linking of Lipases Adsorbed on Aminated Supports in the Presence of Detergents Leads to Improved Performance

et al. 2006

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“…Thus, it is necessary to determine the best detergent and the best concentration for each particular reaction. In a similar way, the effect of the covalent coating of lipases with detergents to be used in organic medium depends on the physical properties of the detergent (Mine et al, 2000(Mine et al, , 2001.…”

Section: Discussionmentioning

confidence: 99%

Improved catalytic properties of immobilized lipases by the presence of very low concentrations of detergents in the reaction medium

Fernández‐Lorente

Palomo

Cabrera

et al. 2006

Biotech & Bioengineering

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The addition of a very small concentration of a detergent (in many instances under the critical micellar concentration (cmc)) has been found to greatly increase the activity of immobilized lipases, using those from Pseudomonas fluorescens (PFL) and Candida antarctica (isoform B) as model enzymes. However, the detergents may also have a negative effect on enzyme activity; in fact, for all enzyme preparations and substrates the activity/detergent concentration curve reached a maximum value and started to decrease, in many instances even under the initial value. The concentration and nature of the detergent (SDS, CTAB, Triton X-100, or X-45) that permitted the maximum hyperactivation was different depending on the substrate. The best hyperactivation values promoted by the presence of detergent were over a 20-fold factor. The presence of detergents permitted the inhibition of lipases by irreversible covalent inhibitors (e.g., 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride) (AEBSF) while the enzyme, in the absence of detergent, is not inhibited by these irreversible inhibitors. This suggested that the main effect of the detergents is to shift the conformational equilibrium of lipases toward the open form. Moreover, the presence of detergents also permitted to improve the enantioselectivity exhibited by the immobilized lipases in some cases. For example, the enantioselectivity of PFL-glyoxyl agarose increased from 40 to more than 100 in the hydrolysis of (+/-)-2-hydroxy-4-phenylbutyric acid ethyl ester by using 0.1% CTAB.

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“…On the other hand, the interaction between surfactants and active site of enzyme should be considered. They would be able to shift the lipase conformation equilibrium toward the open form 27–30. In this sense, surfactants acted as activators for CT.…”

Section: Resultsmentioning

confidence: 99%

Stability and activity of chymotrypsin immobilized on magnetic nanogels covered with carboxyl groups

Jiang

Huang

Liu

et al. 2008

J of Applied Polymer Sci

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a-chymotrypsin as model enzyme was anchored to the carboxyl-functionalized magnetic nanogels prepared by in situ photochemical polymerization. Furthermore, to explore the optimum immobilization, the effects of immobilization time, pH of the reaction mixture, and proportion of enzyme to the magnetic nanogels were studied. The immobilized enzyme was stable in the presence of enzyme denaturation surfactants, and maintained their activity against protein-digesting enzyme. The immobilized enzyme exhibited hydrolysis activity against the substrate of casein, and cleaved the substrate into small fragments.

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Preparation of Detergent-Lipase Complexes Utilizing Water-Soluble Amphiphiles in Single Aqueous Phase and Catalysis of Transesterifications in Homogeneous Organic Solvents.

Cited by 10 publications

References 22 publications

Glutaraldehyde Cross-Linking of Lipases Adsorbed on Aminated Supports in the Presence of Detergents Leads to Improved Performance

Glutaraldehyde Cross-Linking of Lipases Adsorbed on Aminated Supports in the Presence of Detergents Leads to Improved Performance

Improved catalytic properties of immobilized lipases by the presence of very low concentrations of detergents in the reaction medium

Stability and activity of chymotrypsin immobilized on magnetic nanogels covered with carboxyl groups

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