Conformational lability of lipases observed in the absence of an oil-water interface: crystallographic studies of enzymes from the fungi Humicola lanuginosa and Rhizopus delemar.

Derewenda, Urszula; Swenson, Lora; Wei, Yang; Green, R; Kobos, P M; Joerger, Rolf D.; Haas, Michael J.; Derewenda, Zygmunt S.

doi:10.1016/s0022-2275(20)41203-9

Cited by 183 publications

(96 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Lipase hyperactivation exhibited after immobilization on hydrophobic supports is caused by lid movements, exposing the active center of the lipase to the medium, which changes the conformational balance between the open and closed form of the lipase [25][26][27][28][29], and fully displaces the equilibrium towards the open active form of the lipase, which is fixed by adsorption on the hydrophobic support through its hydrophobic pocket [17] (Figure 1). This mechanism facilitates the entry of the substrate, resulting in easier enzymatic hydrolysis [16,17,34,73].…”

Section: Immobilization Of Usba-gbx-513 Lipase On Different Supportsmentioning

confidence: 99%

“…Lipases are the most used enzymes in biocatalysis due to their high activity, specificity, selectivity, and robustness in a variety of reaction media [14][15][16][17][18][19][20][21][22][23][24]. Lipases have a peculiar catalytic mechanism, called interfacial activation, which is based on the interaction of the lipase with hydrophobic structures, which causes deep conformational changes on the structure of the active site of the lipase and its consequent enzyme activation [25][26][27][28][29], permitting the enzyme to act in the hydrolysis of insoluble drops of oils and be adsorbed on any hydrophobic surface [30][31][32][33][34].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Modulation of the Biocatalytic Properties of a Novel Lipase from Psychrophilic Serratia sp. (USBA-GBX-513) by Different Immobilization Strategies

et al. 2021

View full text Add to dashboard Cite

In this work, the effect of different immobilization procedures on the properties of a lipase obtained from the extremophilic microorganism Serratia sp. USBA-GBX-513, which was isolated from Paramo soils of Los Nevados National Natural Park (Colombia), is reported. Different Shepharose beads were used: octyl-(OC), octyl-glyoxyl-(OC-GLX), cyanogen bromide (BrCN)-, and Q-Sepharose. The performance of the different immobilized extremophile lipase from Serratia (ESL) was compared with that of the lipase B from Candida antarctica (CALB). In all immobilization tests, hyperactivation of ESL was observed. The highest hyperactivation (10.3) was obtained by immobilization on the OC support. Subsequently, the thermal stability at pH 5, 7, and 9 and the stability in the presence of 50% (v/v) acetonitrile, 50% dioxane, and 50% tetrahydrofuran solvents at pH 7 and 40 °C were evaluated. ESL immobilized on octyl-Sepharose was the most stable biocatalyst at 90 °C and pH 9, while the most stable preparation at pH 5 was ESL immobilized on OC-GLX-Sepharose supports. Finally, in the presence of 50% (v/v) tetrahydrofuran (THF) or dioxane at 40 °C, ESL immobilized on OC-Sepharose was the most stable biocatalyst, while the immobilized preparation of ESL on Q-Sepharose was the most stable one in 40% (v/v) acetonitrile.

show abstract

Section: Immobilization Of Usba-gbx-513 Lipase On Different Supportsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Modulation of the Biocatalytic Properties of a Novel Lipase from Psychrophilic Serratia sp. (USBA-GBX-513) by Different Immobilization Strategies

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Following these conformational changes, Leu17 in the loop between ␤1 and ␣1 moves to a position near the nucleophile Ser87 and forms the oxyanion hole. As found in R. miehei lipase (RmL) [45], H. lanuginosa lipase (HlL) [30], and the human pancreatic lipase-porcine colipase complex [46], the oxyanion hole in PcL is not preformed but is generated by the opening of the lid. However, the catalytic triads of PcL and PgL in different conformations are virtually superimposable (Fig.…”

Section: The Oxyanion Holementioning

confidence: 99%

“…In the crystal structure of RdL, the two molecules in the asymmetric unit showed different lid conformations [29]: one of them is closed and the other is in an intermediate state between the open and closed forms. All these lipase crystals obtained in the open conformation were grown in the presence of 2-methyl-2,4-pentanediol (MPD) [11,18,30,38]. It seems likely that this specific crystallization medium may exert a positive influence on stabilizing the open conformation of lipases.…”

Section: Lipase Opening and Crystal Packingmentioning

confidence: 99%

See 1 more Smart Citation

The crystal structure of a triacylglycerol lipase from Pseudomonas cepacia reveals a highly open conformation in the absence of a bound inhibitor

et al. 1997

View full text Add to dashboard Cite

. PcL exhibits some structural features found in other lipases. The presence of the Ser-His-Asp catalytic triad, an oxyanion hole, and the opening of a helical lid suggest that this enzyme shares the same mechanisms of catalysis and interfacial activation as other lipases. The highly open conformation observed in this study is likely to reflect the activated form of the lipase at an oil-water interface. The structure suggests that the interfacial activation of bacterial lipases involves the reorganization of secondary structures and a large movement of the lid to expose the active site. This is similar to the mechanism described for other well characterized fungal and mammalian lipases.

show abstract

Lipase-enhanced activity in flavour ester reactions by trapping enzyme conformers in the presence of interfaces

González-Navarro

Braco

1998

Biotechnol. Bioeng.

View full text Add to dashboard Cite

In order to improve the lipase‐catalyzed synthesis of flavour esters, we have used the reported strategy of interfacial activation‐based molecular (bio)imprinting [Mingarro et al. 1995. Proc. Natl. Acad. Sci. U.S.A. 92: 3308], later called trapping in the presence of amphiphile interfaces (TPI) [Mingarro et al. 1996. Biochemistry 35: 9935]. Five lipases of fungal and mammalian origin typically used for esterification process have been explored to improve production by TPI treatment. A marked enhancement of enzymatic activity has been observed in all TPI‐treated lipases assayed and the activation factor obtained was up to 90‐fold. The dependence on chain length of acyl donors in the esterification of geranyl alcohol has been investigated, showing clear differences between activated and nonactivated lipase. The results indicate that this rational approach leads to conversion yields that are remarkably higher, not only than its counterpart pH‐optimized control lipase, but also the “protected” lipase by conventional methods (lyoprotectans or salts). We propose this strategy as a promising tool to be used in more industrial biotransformations. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:122–127, 1998.

show abstract

Conformational lability of lipases observed in the absence of an oil-water interface: crystallographic studies of enzymes from the fungi Humicola lanuginosa and Rhizopus delemar.

Cited by 183 publications

References 20 publications

Modulation of the Biocatalytic Properties of a Novel Lipase from Psychrophilic Serratia sp. (USBA-GBX-513) by Different Immobilization Strategies

Modulation of the Biocatalytic Properties of a Novel Lipase from Psychrophilic Serratia sp. (USBA-GBX-513) by Different Immobilization Strategies

The crystal structure of a triacylglycerol lipase from Pseudomonas cepacia reveals a highly open conformation in the absence of a bound inhibitor

Lipase-enhanced activity in flavour ester reactions by trapping enzyme conformers in the presence of interfaces

Contact Info

Product

Resources

About