Applied Biocatalysis Group
"The power of enzymes"
The Institute of Catalysis and Petrochemistry
This Institute of Catalysis and Petrochemistry is part of the Spanish Council for Scientific Research (CSIC), an autonomous body belonging to the Ministry of Science and Innovation. The Institute is one of the research centres included in the field of Chemistry and Chemical Technologies, which is in turn one of the eight Scientific and Technical Areas of research in which the activity of CSIC is structured.
The main objective of the Institute is to carry out scientific research in the field of catalysis both in chemistry and biology, with special emphasis in the catalyst and the process. The subjects of research deal with refining, petrochemicals, catalysts and processes for the synthesis of commodities and high added value chemicals, the elimination of pollutants or the production and transformation of energy resources. The Institute of Catalysis and Petrochemistry enjoys a strong association, formalised in a cooperative agreement covering teaching and research, with the Autonomous University of Madrid (UAM). the following techniques are available at the Institute: Atomic Emission Spectroscopy (ICP), Atomic Absorption Spectroscopy, FTIR Spectroscopy (Nicolet), UV-VIS Spectroscopy, FT-Raman, RSE XPS. Diffuse Reflectance, N2 Isotherm. B.E.T. Surface, Mercury Porosimetry, Flow Chemisorption, Fluorimetry, X-Ray Diffraction.
Applied Biocatalysis Group
The research activities in biocatalysis at the Institute of Catalysis and Petrochemistry (CSIC) started in 1969 after the return of Dr. Antonio Ballesteros of a postdoctoral training at the National Institutes of Health. During 1974-1990, the group studied the stabilisation of micrococcal endonuclease by insolubilization on polysaccharides (agarose, agricultural wastes) by covalent bonds. Following the oil crisis of 1973, and a sabbatical stay, a new theme was dealt with from 1976 to 1990: the study of hydrogenases, as possible candidates to include in a scheme of biophotolysis of water. Research included induction, handling and purification of oxygen-labile hydrogenases, and their stabilization. In 1987 the study of lipases and esterases was started.
Dr. Francisco J. Plou, from the University of Zaragoza, joined the group in 1989. The presence of organic solvents in the reaction media was exploited for the lipase-catalysed synthesis of compounds used as food additives or surfactants: mono- and diglycerides, and fatty acid esters of di- and trisaccharides. In 1994 the study of enzymes acting on carbohydrates, was initiated. In collaboration with European colleagues, the activity of two glycosyltransferases -cyclodextrin glucosyltransferase (CGTase) and dextransucrase- in the formation of cyclodextrins and prebiotic oligosaccharides was studied. Different immobilisation techniques were tested: entrapment in alginate, covalent attachment to acrylic polymers (Eupergit, Sepabeads, Dilbeads) or activated silica, etc.
Furthermore, a new research topic (directed evolution of enzymes) started 4 years ago in the Applied Biocatalysis Group, coordinated by Dr. Miguel Alcalde, who made a postdoctoral stay in Caltech under the supervision of Prof. Frances Arnold (Pasadena, USA). Directed Molecular Evolution is a new tool in protein engineering that is meaning a revolution in biotechnology. This methodology has been employed to design enzymatic functions never required in natural environments. A series of evolved laccases and peroxidases have been obtained in the last few years in our laboratories.
The group has a strong collaboration with various groups in Spain, such as Centro de Investigaciones Biológicas (Drs. María Jesus Martinez, Angel T. Martínez and Jesús Jiménez-Barbero), Centro de Biología Molecular (Dr. María Fernández-Lobato), Instituto de Investigaciones Químicas y Ambientales de Barcelona, (Drs. José L. Parra y Francisco Comelles), Institute of Organic Chemistry (Dr.. Manuel Bernabé), Departamento de Microbiología de la Universidad de Alcala de Henares (Prof. José L. Copa-Patiño). In addition, the Group maintains a fruitful collaboration with numerous European Laboratories, as a consequence of our participation in several EU Projects.
The Group was in charge of the organization of the EUCHEM Conference “Enzymatic & Homogeneous Catalysis”, Santander, Spain, July 1975. The Group was honoured in 1995 with the "III AIQS Award on Enzyme Technology", sponsored by the Instituto Químico de Sarriá, for the work "Enzymatic acylation of sucrose as a tool to produce potential surfactants". The Group organized in March 1997 a Fundación Juan March Workshop on “Novel Biocatalysts” (Madrid). The Group was in charge of the organization of the Symposium "Stability and Stabilization of Biocatalysts", Córdoba, Spain, April 19-22, 1998 and the “International Symposium on Environmental Biocatalysis” held in Cordoba, from 23th to 26th April 2006.
The Applied Biocatalysis Group has developed a fruitful training activity, namely focused in 15 Doctoral Thesis and 3 Master Thesis. In addition, 5 foreign postdoctoral researchers (2 from Mexico, 1 from England, 1 from India and 1 from Iran) have performed long scientific stays (> 1 year) in our laboratories.
The Applied Biocatalysis Group in Madrid has experience in most of the techniques that are relevant in the field of Biocatalysis:
- Different methodologies for the determination of enzyme activity, e.g. fluorimetric, spectrophotometric and pH static techniques, HPLC.
- Molecular biology techniques.
- High-throughput screening methods.
- Methodology for the immobilization/stabilization of enzymes.
- Chromatographic techniques for purifying and characterizing native and modified enzymes (gel filtration, electrophoresis, FPLC, isoelectrofocusing) and for following the progress of the enzymatic reaction (TLC, HPLC, gas-liquid chromatography).
