Instituto de Biotecnologia UNAM

Grupo del Dr. Francisco Xavier Soberon

Protein directed evolution

Our research is conducted in the context of a Group consortium (with the direct participation of Enrique Morett, Lorenzo Segovia and Alejandro Garciarrubio, and with close links with Eduardo Horjales and Juan Carlos Almagro). Each of these investigators conducts his own projects and also converges on certain projects that have been coordinated within a Program Project, financed by Conacyt (1997-2001). The central concept in the group revolves around the study of protein molecular evolution and the establishment of new methodologies related to them. We are convinced that the predictive capacity around the sequence-structure-funtion relationship in proteins is, and will remain for several years, very limited. However, this a central problem, with large scientific and technological repercussions.

It is also very clear that the evolutionary process, based on variation and selection, that gave rise on the first place to the extraordinary dviersity of natural extant proteins can be analyzed to extend, in the laboratory, the functions of those same proteins. Such an approach is known today as directed evolution. The basic elements to build an enabling technology on directed evolution are, on the one hand, mutagenesis methods, where we utilize those based on PCR, (including Gene Shuffling and STEP protocols) and those based on synthetic DNA (specifically we have developed methods that afford codon-level mutagenesis). Likewise, we extend our capacity to generate molecular diversity with other combinatorial methods that circunvent the limitation imposed by tranformation efficiency. We have also developed systems for the selection of proteins with the desired attributes, specially those based on bacterial strains with specific genes deleted. The methodological elements mentioned above have been applied to study various enzyme model systems, such as beta-lactmase, EcoRI endonuclease, triose phosphate isomerase and the enzymatic activities phosphoribosyl anthranylate isomeras and fructose biphosphate aldolase. Further, this enabling technology is also useful to tackle practical problems in biocatalysis, among which we have been involved with penicillin acylase (useful in the production of semi-synthetic penicillins) and alpha-amylase (which is central in the enzymatic process to obtain high-fructose corn syrup).

The current interest of the group centers around the study of basic concepts behind the process of molecular evolution, based on hypotheses emerging from our own results, such as the role of insertions and deletions, the participation of structural modules and the concepts of flexibility and generality of catalysis in primigenial enzymes. Likewise, we have initiated the application of concepts and methods to develop biocatalysts specifically adapted for processes in the production of aromatic compounds, within a Conacyt-financed grant on emerging areas entitled "Cellular Engineering".

Sources of funding: CONACyT (NC230); DGAPA/UNAM (IN223199)

Research Lines: Development and consolidation of Molecular Biology methods Bioinformatics
Structure, function and manipulation of peptides and proteins

Dr. Francisco Xavier Soberon
Investigador
Tutor de Maestría y Doctorado
Dr. Joel Osuna
Investigador
Dr. Humberto Flores
Técnico Académico
I.B.Q. Alejandra Aquino
Estudiante
Ing. Omar Fernando Cruz
Estudiante
David Galeana
Estudiante
Mariana Gutiérrez
Estudiante
Luis Moises Ledezma
Estudiante
M.C. Brenda Georgina Uribe
Estudiante
M.C. Diego Eloyr Navarro
Estancia Temporal
German A. Uribe
Auxiliar de Laboratorio


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