Work could result in national enzymes as biocatalysts for drug production
Groups dedicated to so-called “green chemistry” are increasingly spreading around the world. In general terms, it can be understood as the creation, development and application of chemical products and processes that aim to reduce or even eliminate the use and generation of substances that are harmful to humans and harmful to the environment. This is one of the aspects of research guided by professor Anita J. Marsaioli, from the Institute of Chemistry (IQ) at Unicamp, linked to the Center of Excellence for Research in Sustainable Chemistry (CERSusChem), based at UFSCar, and in which researchers from Unesp, USP and UFSC. The Center, which aims to research new technologies and solutions for the chemical and pharmaceutical industry, has financial support from Fapesp and the English chemical company GlaxoSmithKline (GLK).
Biologist José Matheus C. Bonatto, graduated from the Escola Superior de Agricultura Luiz de Queiroz (ESALQ-USP), with a master's degree in genetics and plant breeding from the same institution, and a doctorate in biochemistry and molecular biology from the USP Chemistry Institute, was selected, among more than 70 applicants, at the beginning of 2017. The researcher was awarded a post-doctoral scholarship, offered by Fapesp through the Center, to carry out studies with enzymes, intended for the production of drugs and other molecules of interest industry, at the Biocatalysis and Natural Products Laboratory of the Department of Organic Chemistry at IQ at Unicamp, with guidance from the professor.
From then on, given the Center's interests and objectives, he focused on researching Brazilian enzymes and microorganisms that could be used as catalysts in drug synthesis reactions in order to make them faster, cheaper and more sustainable, that is, without generating waste that impacts the environment.
According to Matheus, biocatalysis, which involves the use of enzymes and microorganisms to carry out a chemical reaction, is very important because these living beings allow specific chemical transformations to be carried out in a mild way, without the use of organic solvents and strong acids and relatively high temperatures. This is not always possible with the use of conventional catalysts, already used in the synthesis of pharmaceuticals, which generate waste that requires treatment for disposal in the correct way. “My research project aims to identify and apply enzymes from Brazilian microorganisms with a view to their possible application in the synthesis of drugs”, says the researcher.
Indeed, the main objective of the studies carried out by the Biocatalysis and Natural Products Laboratory is to identify enzymes from all Brazilian biodiversity, characterize them and verify possible applications in the pharmaceutical and chemical industry. In this search, the researcher identified two enzymes in the Brazilian microorganism Neopestalothiopsis sp., called Monoamine Oxidase (MAO-N) and Imine Reductase (IRED), of which there are very few descriptions in the literature and, until then, the DNA sequence was not known. These enzymes were selected by homology, as they turned out to be 86% identical to others already used in drug synthesis and obtained in other countries. As a result, there is a high probability that they will serve the same purpose. But this can only be confirmed in bench tests, as, as the author says, it is a trial and error process. The employment of one of them, MAO-N confirmed what was expected. Regarding the other, IRED, studies are still ongoing.
Work sequence
Matheus says that, from a library of fungi existing in the laboratory, a previous group of postgraduate students supervised by Professor Anita, found a new fungus that produces enzymes that are rare in nature and that could be useful for obtaining pharmaceuticals. As almost nothing was known about its genome, all of its DNA had to be sequenced, which was done in facility from Unicamp (LaCTAD). By analogy with the genomic sequence of foreign fungi, he located enzymes that were of interest within their genome and cloned these sequences into bacteria. E. coli in order to increase the speed of its reproduction.
This is because bacteria are much simpler microorganisms, with much smaller genomes, which facilitate study, and replicate enzymes exponentially in a short time. Because of this, the fungal sequence of interest is placed inside a bacterium that functions as a biofactory, producing immense amounts of the enzyme in a short time. His objective was then to test these enzymes on substrates that contain different molecules that can give rise to drugs, which would allow him to see in which reactions it actually acts. This part of the work is still in progress.
“To obtain a certain drug, it is necessary to bring together certain molecules. We know its precursors, as we already know in advance which molecules must be put together to give rise to a given drug. We therefore have a preliminary view of where this enzyme can act, but it is necessary to test the viability of each of the various possibilities proposed”, explains the researcher. His expectation is to generate, by the end of his post-doctorate, at least the synthesis of one drug in a more sustainable way.
Conference in the USA
Matheus credits, in addition to analyzing his merit and CV, the discovery of this new fungus, the fact that he was awarded a full exemption scholarship – registration, travel, accommodation, food – which enabled his participation in the Gordon Research Conference on biocatalysis, held in July at the University of New England (Biddefor, USA), where more than 350 prominent researchers in the field of biocatalysis from the USA and Europe are present. Regarding Gordon's interest in paying for her participation, which is not cheap, as registration alone costs US$ 1, she says: “A large part of the uses of enzymes of which ours is homologous are intended for the pharmaceutical industry. We brought another fungus with the possibility of this use.”
He also says that his poster, presented over three days, aroused particular interest among American and European researchers for bringing to the attention of a new fungus that expresses an important enzyme for “green chemistry” and that could have relevant pharmaceutical applications. Although it was his first international experience, his impression, in relation to the scientific relevance of the conference, is that there is no significant lag in relation to what is being done in Brazil in the area. So much so that in the poster category his work was selected among the five best.
