Thermomyces lanuginosus lipase

Other microbial lipases have also been successfully used in anhydrous ionic liquids, e.g., from Alcaligenes sp. (AsL) [54, 58], CaLA, Rhizomucor miehei lipase (RmL), and Thermomyces lanuginosus lipase (TIL) [54]. The lipase from pig pancreas (porcine pancreas lipase, PPL), the only mammalian lipase that has been subjected to ionic liquids, catalyzed transesterificationin[BMIm][NTf2]butnotin[BMIm][PF6]... 

TIL Thermomyces lanuginosus lipase, RdL Rhizopus delemar lipase, RnL Rhizopus niveus lipase, MmE Mucor miehei esterase, PsL Pseudomonas sp. lipase, MmL Mucor miehei lipase, RoL Rhizopus orvzae lipase, CaLA Candida antarctica lipase A, CaLB Candida antarctica lipase B, PLE Pig liver esterase, EP Enteropeptidase, PKA Porcine kidney acylase, CE Cholesterol esterase Figure 8.1 (S)-Selective enzyme hits from hydrolase screening. ... 

As an alternative to the chemical resolution methods described by Atwal et al. (Scheme 4.13), a biocatalytic strategy towards the preparation of enantiopure (R) and (S)-SQ 32,926 was developed (Scheme 4.15). The key step in the synthesis is the enzymatic resolution of an N3-acetoxymethyl-activated dihydropyrimidone precursor by Thermomyces lanuginosus lipase [189]. The readily available racemic DHPM 43 was hydroxymethylated at N3 with formaldehyde, followed by standard acetylation with acetyl chloride. The resulting N3-acetoxymethyl-activated DHPM... 

Three different variants of Thermomyces lanuginosus lipase (TLL) were used as the model enzyme to study the hydrolysis of the phopsholipid bilayer. Being a lipase, TLL has low affinity for phospholipid bilayers [39,40]. TLL is, therefore, of interest for the characterization of the diffusion and adsorption... 

The application of ionic liquids in lipase biocatalysis has not remained entirely restricted to CaLB, PcL or CrL. Other lipases have been used in ionic liquids for ester synthesis such as Candida antarctica lipase A (CaLA) [15,16], Thermomyces lanuginosus lipase [17] (TLL), Rhizomucor miehei lipase (PmL), Pseudomonas fluorescens lipase (PJL) [18], Pig pancreas lipase (PpL) [17] and Alcaligenes sp. lipase (A5 L) [16]. 

Chirazyme L2-C2 (CAL-B) proved to be a very useful enzyme for the development of an acylation process for the large-scale production of vitamin A (retinol, 91) at Roche (Scheme 27) [90,91]. In the plant process of vitamin A, intermediate 88 is partially acylated and then subjected to acid-catalyzed dehydration and isomerization to yield the vitamin A ester 90 via acetate 89. Contrary to the chemical acylation, an enzymatic approach allowed for a highly selective monoacylation of 88, and Chirazyme L2-C2 showed a very high conversion rate at 30% (w/w) substrate concentration. A first continuous process on the laboratory scale was set up with a 15 ml fixed-bed reactor containing 5.0-8.0 g of immobilized biocatalyst 4.9 kg of 89 was synthesized within 100 days in 99% yield and with 97% selectivity for the primary hydroxyl group. The laboratory process was implemented in a miniplant (120 g of biocatalyst), which could convert 1.4 kg of 88 into 1.6 kg 89 per day. After 74 days the conversion efficiency was still 99.4%. Further development of this transformation led to a modified process, which uses Thermomyces lanuginosus lipase immobilized on Accurel MPlOOl for the continuous production of 89 [92]. 

We initially selected methyl caffeate and 3-cyclohexyl-l-propanol as substrates for comparative study of the enzyme s performance in [bmim][NTf2]. Four commercially available lipases, C. antarctica lipase B (Novozyme 435, Novozymes, 30 U mg-i), Rhizotnncor miehei lipase (RMIM, Novozymes, 1370 U mg-i), B. cepacia lipases (PS-CI, Wako Pure Chemical, Osaka, Japan, 2560 U mg-i), and Thermomyces lanuginosus lipase (TLIM, Novozymes, 1850 U mg-i) were tested (Table 2). 

In addition to cutinases, various lipases, such as from C. antarctica, Candida sp. [13, 47], Thermomyces lanuginosus [2, 14, 15, 55, 56], Burkholderia (formerly Pseudomonas) cepacia [57] and esterases from Pseudomonas sp. (serine esterase) [58] and Bacillus sp. (nitrobenzyl esterases) [59, 60], have shown PET hydrolase... 

High molecular weight poly(trimethylene carbonate) PTMC (Mn 300 k g/mol, Mw/Mn 1.46), synthesized, purified, and characterized as described in reference [72] is dissolved in chloroform (3 mg/mL). Thin films are prepared by spin-coating these solutions on cleaned Si wafers at 3,000 rpm (film thickness obtained 25 50 nm). Film thicknesses can be determined by AFM imaging using the scratch method described in Chap. 2 (see also above, hands-on example 47). The enzymatic reaction takes place in situ in the liquid cells filled with lipase solutions (lipase from Thermomyces lanuginosus (EC3.1.1.3, minimum 50,000 units/g purchased from Sigma, U.S.A.) at 37°C for 30 s, 1 min, and 2 min, respectively. 

In this work, MG and DG are produced through lipase-catalyzed glycerolysis of soybean oil in a batch reactor using Candida antarctica B, Thermomyces lanuginosus, Rhizomucor miehei, Candida rugosa, and Aspergillus niger lipases, in a solvent-free system. 

Among spectroscopic methods, X-ray photoelectron spectroscopy (XPS), also known as electron spectroscopy for chemical analysis (ESCA), has been used for the analysis of enzyme-treated PET substrates [11, 27, 31, 102]. With XPS, the elemental composition of the top 10 nm of the PET surface is measured, and the presence of specific functional groups can be identified. XPS analysis of PET treated with a lipase from Thermomyces lanuginosus and a cutinase from Ther-mobifida fusca indicated increased amounts of hydroxyl and free acid groups, while the amount of carbon-carbon bonds decreased [11, 27], XPS has been shown to be very sensitive to contamination by residual enzyme protein strongly adsorbed to the polymer surface [31, 102]. 

Fernandez-Lafuente R (2010) Lipase ftom Thermomyces lanuginosus. Uses and prospects as an industrial biocatalyst. J Mol Cat B Enzym 62 197-212... 

PPT) were treated with polyesterases from Thermomyces lanuginosus, Penicillium citrinum, Thermobifida fusca and Fusarium solani pisi, and the cutinase from Thermobifida fusca was found to release the highest amounts of hydrolysis prodncts from PPT materials [37]. The Thermobifida fusca enzyme hydrolysed both PPT fibres and films, whereas the lipase from Thermomyces lanuginosus was only able to hydrolyse the fibres. Due to the higher surface area, fibres are more easily attacked by enzymes than films. 

Ferrer, M., Plou, F. J., Puentes, G. et al. (2002) Effect of the immobilization method of lipase from Thermomyces lanuginosus on sucrose acylation. Biocatal. Biotransform., 20, 63-71. 

Until now, thoroughly investigated commercial immobilized lipases are Novozym 435 (Hemandez-Martin and Otero, 2008), Lipozyme TLIM (Wang et al., 2008), and Lip-ozyme RM IM (Aguieiras et al., 2013). AU of them are extracellular enzymes. The most widely used are Novozym 435, from Candida antarctica, immobilized on a mac-roporous acryhc resin Lipozyme RM IM, from Rhizomucor miehei, immobilized on an anionic resin and Lipozyme TL IM, from Thermomyces lanuginosus, immobilized on a gel of granulated sUica. 

Verdugo, C., Luna, D., Posaddlo, A., Sancho, E.D., Rodriguez, S., Bautista, F., Luque, R., Marinas, J.M., Romero, A.A., 2011. Production of a new second generation biodiesel with alow cost lipase derived fiom Thermomyces lanuginosus optimization by response surface methodology. Catalysis Today 167, 107—112. 

Abstract The functionalization of synthetic polymers such as poly(ethylene terephthalate) to improve their hydrophilicity can be achieved biocatalytically using hydrolytic enzymes. A number of cutinases, lipases, and esterases active on polyethylene terephthalate have been identified and characterized. Enzymes from Fusarium solani, Thermomyces insolens, T. lanuginosus, Aspergillus oryzae, Pseudomonas mendocina, and Thermobifida fusca have been studied in detail. Thermostable biocatalysts hydrolyzing poly(ethylene terephthalate) are promising candidates for the further optimization of suitable biofunctionalization processes for textile finishing, technical, and biomedical applications. 

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