The lipase from Thermomyces laguginosus (formerly Humicola laguginosa) (TLL) is a basophilic and noticeably thermostable enzyme, commercially available in both soluble and immobilized form. Although initially oriented toward the food industry, the enzyme has found applications in many different industrial areas, from biodiesel production to fine chemicals (mainly in enantio and regioselective or specific processes). This review intends to show some of the most relevant aspects of the use of this interesting enzyme. After checking the enzyme features, some of the most efficient methods of TLL immobilization will be commented. Finally, the main uses of the enzyme will be revised, with special emphasis in the modification of fats and oils, production of biodiesel, resolution of racemic mixtures, enantioselective hydrolysis of prochiral esters and regioselective process involving sugar preparations. In many instances, TLL has been compared to other lipases, the advantages or disadvantages of the enzyme will be discussed.

Lipase from Thermomyces lanuginosus: Uses and prospects as an industrial biocatalyst

The lipase from Rhizomucor miehei (RML), formerly Mucor miehei, is a commercially available enzyme in both soluble and immobilized forms with very high activity and good stability under diverse experimental conditions (anhydrous organic solvents, supercritical fluids, etc.). The uses of the enzyme were initially oriented towards food industry, that way the enzyme has found a broad application in this area. This review intends to show the enzyme features and some of the most relevant aspects of the use of this interesting enzyme in oils and fats modification. The enzyme has been the first lipase whose structure was reported and its interfacial activation is very well known, making this enzyme a good template for modeling studies. Finally, the main uses of the enzyme in fatty acids, oils and fats modification will be revised (hydrolysis of glycerides, transesterification, esterification, acidolysis and interesterification). This lipase seems to be advantageous when compared to other lipases mainly in esterificaton reactions in anhydrous media, while other lipases can be preferred in transesterification reactions.

Lipase from Rhizomucor miehei as a biocatalyst in fats and oils modification

With current advances in genomics, several technological processes have been generated, resulting in improvement in different segments of molecular research involving prokaryotic and eukaryotic systems A widely used contribution is the identification of new genes and their functions, which has led to the elucidation of several issues concerning cell regulation and interactions For this, increase in the knowledge generated from the identification of promoters becomes considerably relevant, especially considering that to generate new technological processes, such as genetically modified organisms, the availability of promoters that regulate the expression of new genes is still limited Considering that this issue is essential for biotechnologists, this paper presents an updated review of promoters, from their structure to expression, and focuses on the knowledge already available in eukaryotic systems Information on current promoters and methodologies available for studying their expression are also reported

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Plant promoters: an approach of structure and function.

Cotton continues to be a crop of great economic importance in many developing and some developed countries. Cotton plants expressing the Bt gene to deter some of the major pests have been enthusiastically and widely accepted by the farmers in three of the major producing countries, i.e., China, India, and the USA. Considering the constraints related to its production and the wide variety of products derived from the cotton plant, it offers several target traits that can be improved through genetic engineering. Thus, there is a great need to accelerate the application of biotechnological tools for cotton improvement. This requires a simple, yet robust gene delivery/transformant recovery system. Recently, a protocol, involving large-scale, mechanical isolation of embryonic axes from germinating cottonseeds followed by direct transformation of the meristematic cells has been developed by an industrial laboratory. However, complexity of the mechanical device and the patent restrictions are likely to keep this method out of reach of most academic laboratories. In this chapter, we describe the method developed in our laboratory that has undergone further refinements and involves Agrobacterium-mediated transformation of cotton cells, selection of stable transgenic callus lines, and recovery of plants via somatic embryogenesis.

Cotton (Gossypium hirsutum L.).

Plant sterols have attracted increasing attention due to their excellent cholesterol-lowering activity. However, free plant sterols have some characteristics of low oil solubility, water insolubility, high melting point, and low bioavailability, which greatly limit their application in foods. Numerous studies have been undertaken to modify their chemical structures to improve their chemical and physical properties in meeting the needs of various applications. The present review is to summarize the literature and update the progress on structural modifications of plant sterols in the following aspects: (i) synthesis of plant sterol esters by esterification and transesterification with hydrophobic fatty acids and triacylglycerols to improve their oil solubility, (ii) synthesis of plant sterol derivatives by coupling with various hydrophilic moieties to enhance their water solubility, and (iii) mechanisms by which plant sterols reduce plasma cholesterol and the effect of structural modifications on plasma ch...

Plant Sterols: Chemical and Enzymatic Structural Modifications and Effects on Their Cholesterol-Lowering Activity.

Structured phospholipids were synthesized with the functional lipid conjugated linoleic acid (CLA). The lipase- and phospholipase A 2 -catalyzed enzymatic acidolysis reaction between phospholipids (PL) and CLA was used for fatty acid modification. Enzymatic processes were an effective way to produce structured PL. Screening of four lipases and immobilized phospholipase A 2 and a combination of lipase and phospholipase showed that only Lipozyme RM IM and Lipozyme TL IM were effective in incorporation of CLA into PL. The maximum incorporation achieved by the latter enzyme was 16% with soy PL in 72 h.

Enzyme‐catalyzed synthesis of structured phospholipids with conjugated linoleic acid

In the present study, plant sterols were modified to form amphiphiles by synthesizing phospholipid derivatives from them so they could be formulated in different functional foods and possibly improve their effects as therapeutic agents. The new phosphatidyl derivatives, phosphatidyl-sitosterols, were synthesized by the transfer reaction of a phosphatidyl residue from PC to β-sitosterol by phospholipase D (PLD EC 3.1.4.4) from Streptomyces sp. in a biphasic medium.

Phospholipase D-catalyzed synthesis of novel phospholipid-phytosterol conjugates

Lipase-catalyzed acidolysis of different TAG with CLA was performed to produce structured lipids (SL) containing CLA. An immobilized lipase from Mucor miehei (Lipozyme IM, Novo Nordisk Inc., Franklinton, NC) was used as the biocatalyst in a solvent-free system. Conconut oil and tricaprylin, which are sources of medium-chain FA, were the starting substrates, and a mixture of FFA (MFFA) containing 73% CLA was the donor of the acyl groups. For each TAG, four different ratios of TAG/MFFA were blended to prepare about 500 g of mixture containing 10, 20, 30, and 40% CLA (w/w). Each blend was reacted with 5% lipase at 65°C for 48 h under nitrogen. Over the range of TAG/MFFA ratios examined, CLA was incorporated effectively by the enzyme. Lipozyme IM exhibited no special preference for any particular FA, since the incorporation of FA was proportional to their concentration in the system. FFA, PV, p-anisidine value (p-AV), iodine value (IV), and saponification number (SN) were evaluated for all the SL. FFA, PV, and p-AV depended on the purification process and showed no significant deterioration of SL with respect to the original TAG, whereas IV and SN depended on the composition of the SL, mainly the CLA content.

Solvent-free enzymatic synthesis of structured lipids containing CLA from coconut oil and tricaprylin

As opposed to first-generation biotechnology products, such as pest-resistant crops and herbicide-resistant crops, second-generation products often utilize plant-derived, homologous or heterologous genes and/or promoters. In this study, we evaluated the ability of a promoter from a gene encoding a major storage protein in cottonseed to drive an antisense sequence of the cotton FAD2 gene to down-regulate the activity of Delta-12 desaturase enzyme in cottonseeds. The oleic acid level in the transgenic cottonseeds approximately doubled from the wild-type level of 15%, with a concomitant decrease in the level of linoleic acid. A more extensive study of one line revealed a higher degree of seed-to-seed variability in the transgenic phenotype. A thorough investigation was conducted to determine the impact of the use of a homologous promoter to drive a transgene on the activity of the endogenous promoter. The results showed that the use of the homologous alpha-globulin B promoter for transgenic purposes did not adversely affect the expression of alpha-globulin B storage protein in cottonseed. The results obtained in this investigation on the use of a homologous promoter and antisense technology will be useful in the design of strategies to alter biosynthetic pathways for nutritional quality improvements and for the production of heterologous proteins of commercial value in seeds.

A comprehensive study of the use of a homologous promoter in antisense cotton lines exhibiting a high seed oleic acid phenotype.

Lipid nutritional supplements have been in use before the term nutraceutical was coined. Products such as fish oils, shark cartilage, shark liver oil, and vitamins have been in the market since the beginning of the twentieth century. Some of the health claims of these products lacked strict scientific documentation in the past, and their curative properties were mostly anecdotal. However, today there is a better understanding of the biological properties of lipids and their application has extended to combined pharmaceutical and cosmetic fields such as disease prevention and treatment, excipients and coadjuvants, trans-dermal carriers, and skin emolliency agents. This has led to the development of bioactive cosmetic and pharmaceutical products whose name has recently been coined as cosmeceuticals.


Keywords:

structured lipids;
cosmeceuticals;
nutraceutical;
omega-3 fats;
fat-soluble vitamins;
sterols;
steroids;
phospholipids;
trans-dermal carriers

Ernesto Hernandez

Papers

Enzyme‐catalyzed synthesis of structured phospholipids with conjugated linoleic acid

Phospholipase D-catalyzed synthesis of novel phospholipid-phytosterol conjugates

Solvent-free enzymatic synthesis of structured lipids containing CLA from coconut oil and tricaprylin

A comprehensive study of the use of a homologous promoter in antisense cotton lines exhibiting a high seed oleic acid phenotype.

Pharmaceutical and Cosmetic Use of Lipids