High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity

In this study, bovine lactoferrin (bLf), an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf) in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin) demonstrated antibacterial activity against Escherichia coli (E. coli) BL21DE3, Staphylococcus aureus (S. aureus) FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa) ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly

Chloroplasts: The Future of Large-Scale Protein Production

Chloroplast engineering has matured considerably in recent years. It is emerging as a promising tool to address the challenges related to food security, drug production, and sustainable energy posed by an ever-growing world population. Chloroplasts have proven their potential by efficiently expressing transgenes, encapsulating recombinant proteins, and protecting them from cellular machinery, making it possible to obtain highly functional proteins. This quality has also been exploited by interfering RNA technology. In addition to the practical attributes offered by chloroplast transformation, such as the elimination of position effects, polycistronic expression, and massive protein production, the technique represents an advance in biosafety terms; however, even if its great biotechnological potential, crops that have efficiently transformed are still a proof of concept. Despite efforts, other essential crops have remained recalcitrant to chloroplast transformation, which has limited their expansion. In this chapter, we address the most recent advances in this area and the challenges that must be solved to extend the transformation to other crops and become the de facto tool in plant biotechnology

Biocombustibles: estrategias limpias para combatir la crisis energética

México se encuentra ante una eventual crisis energética debido a la reducción en sus reservas probadas del petróleo, lo que ha tenido como consecuencia un incremento en los precios de los combustibles. Además, la utilización de este hidrocarburo ha generado emisión de gases con efecto invernadero, contribuyendo al cambio climático. Para solucionar este problema se requiere desarrollar tecnologías alternativas que nos permitan sustituir los combustibles derivados del petróleo. Estos hechos hacen evidente la necesidad de utilizar fuentes alternas de energía. Los biocombustibles son recursos energéticos producidos por el ser humano a partir de materias generadas por seres vivos, a las cuales se les denomina “biomasa”. Esta segunda generación de biocombustibles plantea el uso de lignocelulosa, que es el polímero más abundante sobre la superficie del planeta; para lograrlo se requiere del desarrollo biotecnológico que permita la despolimerización efectiva de la biomasa vegetal. Abstract Mexico is facing a possible energy crisis due to the reduction in proven reserves of oil and consequently, it has resulted in the increase in fuel prices. To solve this problem it requires developing alternative technologies that allow us to replace fuels from petroleum. Besides that, the use of this fuel has generated greenhouse gases emissions, contributing to the climate change. These facts make clear the need for alternative energy sources. Biofuels are a type of fuel whose energy is derived from living organisms called “biomass” and produced by human beings. These second-generation biofuels require the use of lignocellulose that is the most abundant polymer on the surface of the planet, to get this, it is required the development of a biotechnology process that allows the effective depolymerization of plant biomass

Chemical Characterization and Enzymatic Control of Stickies in Kraft Paper Production

Paper recycling has increased in recent years. A principal consequence of this process is the problem of addressing some polymeric components known as stickies. A deep characterization of stickies sampled over one year in a recycled paper industry in México was performed. Based on their chemical structure, an enzymatic assay was performed using lipases. Compounds found in stickies by Fourier-transform infrared spectrometry were poly (butyl-acrylate), dioctyl phthalate, poly (vinyl-acetate), and poly (vinyl-acrylate). Pulp with 4% (w/w) consistency and pH = 6.2 was sampled directly from the mill once macrostickies were removed. Stickies were quantified by counting the tacky macrostructures in the liquid fraction of the pulp using a Neubauer chamber before the paper was made, and they were analyzed with rhodamine dye and a UV lamp. Of the two commercial enzymes evaluated, the best treatment condition used Lipase 30 G (Specialty Enzymes & Biotechnologies Co®, Chino, CA, USA) at a concentration of 0.44 g/L, which decreased 35.59% of stickies. SebOil DG (Specialty Enzymes & Biotechnologies®) showed a stickies reduction of 21.5% when used at a concentration of 0.33 g/L. Stickies in kraft paper processes were actively controlled by the action of lipases, and future research should focus on how this enzyme recognizes its substrate and should apply synthetic biology to improve lipase specificity