Lipases are enzymes that catalyze the hydrolysis of carboxylic esters at a lipid–water interface and are able to catalyze reactions such as alcoholysis, esterification, transesterification, and enantioselective synthesis in organic media. They are important biocatalysts for biotechnological and industrial applications—such as in the food and flavor industry—and in the production of biopharmaceuticals, biofuels, biopolymers, and detergents. A desirable property of lipases is stereoselectivity for the production of chemicals with high optical purity. In this work, we report the improvement of the enantioselective capabilities of the Geobacillus thermoleovorans CCR11 lipase. By means of a rational design and bioinformatic approaches, six amino acids of the catalytic cavity of the lipase LipTioCCR11 were substituted resulting in an increase in the optimum temperature of the enzyme and in the resistance to the presence of organic solvents in hydrolytic reactions, and in the promotion of the enantioselective recognition of R isomers of carboxylic acids with importance for the pharmaceutical and food industries.

Immobilization of enzymes has many advantages for their application in biotechnological processes. In particular, the cross-linked enzyme aggregates (CLEAs) allow the production of solid biocatalysts with a high enzymatic loading and the advantage of obtaining derivatives with high stability at low cost. The purpose of this study was to produce cross-linked enzymatic aggregates (CLEAs) of LipMatCCR11, a 43 kDa recombinant solvent-tolerant thermoalkaliphilic lipase from Geobacillus thermoleovorans CCR11. LipMatCCR11-CLEAs were prepared using (NH₄)₂SO₄ (40% w/v) as precipitant agent and glutaraldehyde (40 mM) as cross-linker, at pH 9, 20 °C. A U₁₀(5⁶) uniform design was used to optimize CLEA production, varying protein concentration, ammonium sulfate %, pH, glutaraldehyde concentration, temperature, and incubation time. The synthesized CLEAs were also analyzed using scanning electron microscopy (SEM) that showed individual particles of <1 µm grouped to form a superstructure. The cross-linked aggregates showed a maximum mass activity of 7750 U/g at 40 °C and pH 8 and retained more than 20% activity at 100 °C. Greater thermostability, resistance to alkaline conditions and the presence of organic solvents, and better durability during storage were observed for LipMatCCR11-CLEAs in comparison with the soluble enzyme. LipMatCCR11-CLEAs presented good reusability by conserving 40% of their initial activity after 9 cycles of reuse.

Alternative therapies for cancer treatment have been developed using bioactive compounds such as betulinic acid (BA). The objective of this study was to investigate the bioactivity of BA in its free form and compare it with its nano-encapsulated form under a skin carcinogenesis protocol in a genetically modified murine model. K14E6 and FVB mice were divided into four groups to be treated with free BA and with betulinic acid nanoemulsion (BANE). Lecithin enriched with medium chain fatty acids (MCFAs) was employed as an emulsifier to prepare the nanoemulsions with a mean droplet size of 40 nm. Skin tumors were induced by exposure to DMBA and TPA directly to the transgenic mice. Tumor development was completely inhibited by BANE and by 70% with free BA. This was validated by histological sections and the gene expression of the Cdk4 and Casp8 genes.

Los Humeros volcanic complex, located at the eastern end of the Trans Mexican Volcanic Belt, possess superficial thermal manifestations (fumaroles, heat steam-soils and alteration zones), with temperatures between 50 °C and 90 °C. This is the first work to explore the bacterial diversity of a steam soil of Los Humeros geothermal field using a metagenomic 16S RNA gene analysis, and the presence of lipolytic enzymes’ genes after an enrichment culture by a whole-genome shotgun approach. High diversity indexes were found in soil samples, with Firmicutes, Chloroflexi, Proteobacteria, Actinobacteria and Acidobacteria as the most prevalent phyla. Alicyclobacillus was the most prevalent genus at 15 cm and Anoxybacillus at 50 cm depth; anaerobic and obligate thermophilic and hyperthermophilic members were found at 50 cm depth. Firmicutes was favored by the oil-rich medium, and the dominant genera were Geobacillus and Caldibacillus. 165 genes related to lipolytic enzymes were found; carboxilesterases and phospholipases C were the most abundant, and 43% were assigned to uncharacterized lipolytic enzymes. The diversity as well as the high percentage of non-characterized lipolytic enzymes found after soil enrichment in an oil-rich medium suggest the presence of biotechnologically promising organisms and enzymes that deserve to be investigated.

The biotechnological applications of chitinases are diverse. They are used in industrial sectors such as pharmaceuticals, textiles, and agriculture, including the use of recombinant chitinases for pest control, since traditional treatments affect and contaminate hive products. Bacillus licheniformis UV01 bacterium is of interest, as it expresses genes for different enzymes, including chitinase. The Chibluv01 gene was cloned into the pHTP8 vector with a His/tag for purification using affinity chromatography. It showed a specific activity of 115 U/m. The optimal pH and temperature were 7.5 and 42 °C, respectively. The choleoptericidal activity (ability to kill beetles) of the enzyme was evaluated in the larvae and adult beetles of Aethina tumida treated with immersion in a purified enzyme extract at different concentrations, and the mortality was verified at 24, 48, and 72 h. Within 24 h of application, the mortality increased by 50% in the larval stage and 56.6% in adult beetles compared to the control groups. The LC₅₀ and LC₉₀ were obtained: 104.05 U/mL and 234.36 U/mL in larvae and 92.99 U/mL and 211.14 U/mL in adults, respectively. These results indicate the potential of the application of ChiBlUV02 chitinase in pest control.

Biocatalysis is the use of enzymes to accelerate chemical reactions and has undeniable advantages over conventional catalysis, since it significantly reduces the use of toxic compounds and solvents, generates less hazardous waste, and allows working in less aggressive conditions of temperature and pH, Despite these benefits, enzymes due to their protein nature can be easily denatured and being soluble, they are not recoverable from the reaction medium, therefore, enzymatic immobilization in a specialty of biocatalysis in constant development and growth since it allows a greater stability, reuse and easy recovery of the medium, its use is already widespread and is increasing in industries such as pharmaceuticals, biofuels, the food industry and the production of flavors, fragrances and cosmetics. The different strategies to obtain immobilized derivatives are addressed in this document. La biocatálisis es el uso de enzimas para acelerar las reacciones químicas y posee innegables ventajas sobre la catálisis convencional, ya que disminuye sensiblemente el uso de compuestos tóxicos y disolventes, genera menos residuos peligrosos, y permite trabajar en condiciones menos agresivas de temperatura y pH, a pesar de estas bondades, las enzimas, por su naturaleza proteica, pueden ser fácilmente desnaturalizadas y al ser solubles, no son recuperables del medio de reacción, por ello, la inmovilización enzimática es una especialidad de la biocatálisis en constante desarrollo y crecimiento ya que permite una mayor estabilidad, reuso y fácil recuperación del medio, su aplicación es ya generalizada y va en aumento en industrias tales como la farmacéutica, la de los biocombustibles, la alimentaria y de producción de sabores, fragancias y cosméticos. Las diferentes estrategias para obtener derivados inmovilizados se abordan en el presente documento. Enzyme immobilization strategies Biocatalysis is the use of enzymes to accelerate chemical reactions and has undeniable advantages over conventional catalysis, since it significantly reduces the use of toxic compounds and solvents, generates less hazardous waste, and allows working in less aggressive conditions of temperature and pH, Despite these benefits, enzymes due to their protein nature can be easily denatured and being soluble, they are not recoverable from the reaction medium, therefore, enzymatic immobilization in a specialty of biocatalysis in constant development and growth since it allows a greater stability, reuse and easy recovery of the medium, its use is already widespread and is increasing in industries such as pharmaceuticals, biofuels, the food industry and the production of flavors, fragrances and cosmetics. The different strategies to obtain immobilized derivatives are addressed in this document.