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Flavio Alterthum

Flavio Alterthum: Lessons about Brazil’s ethanol

Eduardo CesarA few months ago, Professor Flavio Alterthum, 70, co-author of the well known basic microbiology textbook adopted by Brazilian universities, released an illustrated popular science book for children of seven years and older published by Atheneu, called Pai, o que é micróbio? (“Father, what is a microbe?”). The first edition had a print run of 750 copies and sold out quickly. At this point, while continuing to teach graduate level courses at the Biomedical Sciences Institute of the University of São Paulo (USP) and the Jundiaí Medical College, he excitedly plans to work on other books aimed at this young audience breathing new life into his longtime fascination with story-telling.

The dissemination of science to children is just another challenge in the life of this researcher, who is used to making significant career concessions in order to follow his dreams – which no doubt earned him notable successes, but also some very tough setbacks. In the early 1990’s, for example, proud and enthusiastic, he exhibited his then advanced work on genetically modified bacteria to optimize the fermentation process in ethanol production. This work resulted in a patent, number 5 million, that he obtained along with two of his colleagues from the United States of America. This work reverberated widely in major international newspapers, including The New York Times, and in Brazilian publications, such as O Estado de S. Paulo and Gazeta Mercantil. He even made television appearances to discuss his work. But a few years later, you would have found him very disappointed, and perhaps even a little bitter, because his plan to work in the private sector using the new processes he helped to develop for the more efficient production of ethanol, at the invitation of a company that had just established itself in the Brazilian market, had instead been a huge disappointment. The company simply closed its Brazilian subsidiary, under somewhat nebulous arguments, soon after he had announced his retirement from USP to embark on this new project. Professor Alterthum found himself somewhat lost at that moment, his dreams having crumbled.

But as a teacher par excellence, it was his ability to fully absorb the pleasures of being in front of a group of students, lecturing as well as listening, absorbing questions as they shaped the flow of a fascinating story, as he says, that helped him to recover. It was at USP where found his ground again, although he was officially retired. Always restless, he was soon inventing new projects, something he continues to do to this day. He talks about this path of the researcher and teacher, which sometimes draws curious parallels with the past comings and goings of science and technology in Brazil, in the following interview with Pesquisa FAPESP.

After writing the most widely respected microbiology textbook in the country, after four decades of serious research work, you decided to create a book about microbiology for children. Why this book at this point in your life?
“At this point in my life” it goes a long way back … I graduated in 1964. In 1965 I was hired by USP and remain involved there to this day, although I am now retired. I teach two graduate courses. I am also active in the Graduate School of the Jundiaí Medical School. I have worked there for 15 years. But allow me to answer your question. The title of the book asks, “What is a microorganism?”, an idea that has lain dormant since my daughters were 5, 6 years old. There was a little song that spoke of bacilli and, whenever they heard it, they laughed and asked, “Dad, what is a bacillus?” Of course they knew nothing of microbiology. That stuck in my head and, as I have this great dream, which is to be a storyteller – I began to take courses, even getting a certificate – I thought that by doing a children’s book I might have the opportunity to begin telling stories to small groups and make my knowledge available to people.

What was the song that spoke of bacilli in those years?
I can’t remember. My daughters remember the situation, but not the song or who sang it.

How and where did you take the storytelling course?
At USP, about 20 years ago. It was an extension course taught by a professor from Minas Gerais, an expert on Guimarães Rosa, I don’t recall her name. It was interesting because the course changed some of my ideas about storytelling. I always enjoyed telling stories to my daughters, in my own way. And she told the stories of Guimarães Rosa by the book. I told her I didn’t want to be a storyteller with a fixed script, but have the ability to develop, to invent a story on the spot. This is somewhat related to how I give my courses, especially in the graduate school.

So there is a relationship between storytelling and being a graduate school professor? What is the possible connection between these things?
Are you able to look into the eyes of your students or your listeners and see that they are shining. Perhaps many people have this knowledge, but the teacher and actor who can really speak to their audience, both realize when they are captivating. If I can make people fascinated with microbiology, I am very pleased.

What was the magic, that special thing , that the storytelling course gave to you?
The clear notion that we must listen to people and know how to work with the questions and the knowledge they have. When you give an answer that brings together some element that the person already knows, it encourages them to follow along with whatever you are proposing. I’m speaking about interaction, my classes here [at USP] are interactive. And when there are no questions, I ask the questions.

Then, in class, the thread of “storytelling” is the student’s question or your own repertoire of familiar questions.
Perfect. And in the case of a children’s book it was, first, my daughter’s question. Then I was asking the questions to which I wanted to elicit answers and Telma Alves Monezi created the illustrations.

But how was the experience of translating what you normally present to young students to a much younger audience with a smaller repertoire of knowledge?
It demanded a change to do, hear, think, re-do, until reaching a satisfactory point. I have never had the return I have now, so I was very pleased when my 4 year old granddaughter asked me one day: “Hey Grandpa, did you bring the book of pathogens?”. For her, a 4 year old who could not yet read, the pathogen was a drawing of a bacterium with the face of a duck that appears in the book.

How long did you spend in order to get the text as it actually appears in the book?
Two and a half years. The first thing I wanted to do for the public to avail themselves of my reasonable knowledge of microbiology – one of the scientific disciplines in which its applications are often the easiest means to understanding – there was a radio program that would discuss some peculiarity of this area every week. I got to talk about some issues. For example, the gecko, walking on the ceiling doesn’t fall. Do you know why?

Why?
It’s because the gecko has a substance with a very high adhesion capacity on its feet, and many scientific papers have explored this in some way. For example, researchers cloned the gecko’s genes that are responsible for producing this substance, introduced these genes to a very simple bacterium, Escherichia coli, which acts like a machine and begins to produce the substance on a large scale. And there you have it, a natural glue. It is not on the market because during the production stage it turned into a gum so strong that no one has yet succeeded in separating the thread. This is one of many such examples. Another is the spider’s web, a natural substance with a very high capacity to withstand impact. Again, the spider’s genes that make up the synthesis of the protein that is responsible for the impact resistance were inserted into E. coli. NASA was interested because it wanted to create a material that would cushion better against impact, Formula 1 has similar interests, and today we have just such a material that is created in this way. I wanted to create a book, a work illustrating some of these things, I proposed this idea to Laertes [Coutinho, cartoonist], but he said that he had too many commitments, and ultimately it didn’t work out. There began the story of the microbiology book. I like to speculate about the limits of using such knowledge, to experiment with the possibilities of the interactivity of things. It fascinates me!

If there is a potential fascination with scientific knowledge for all ages, why does scientific culture seem to be so lacking among, for example, Brazilian elementary and high school students?
There are not many books available nor has there been successful disclosure of such knowledge in a simple, pleasant manner, since the beginning of people’s schooling. I did not even know microbiology existed before entering university.

EDUARDO CESARYour book ends with the question But Dad, what is sustainability?. So to summarize for our readers, what is the path that the book takes in order to explain to children what a microbe is. In fact, summarize a little bit the idea of the book.
It’s the simplest thing, from questions. So, who are these microbes? Bacteria, fungi, protozoa, algae, viruses … Where do they live? What do they do? Only bad things? Or do they also do good things? And from there I take each group and talk a little bit about it. Tell about what they do in nature, good and bad. Because, in addition to talking about diseases, I also discuss the positive applications of microorganisms: production of food, alcoholic beverages, antibiotics, and fuel, like in the case of ethanol. We explain the importance to ruminant organisms of having a particular microbiota in their rumen (a specialized part of the digestive tract) and so on and so forth.

There is a lot to the notion that we are complex beings, and that all macroorganisms are, in a certain way, like an immense colony of microorganisms. It’s funny that this scares most people is it not?
It’s because this is a little known fact: we have more microorganisms in our bodies than we do cells. A typical adult has 1014 different microorganisms in their gut. Or rather, there are 100 trillion microorganisms in the gut. And that is just talking about bacteria. Viruses don’t even count because it’s not possible to estimate this. While we have 1012cells, or so. We are made up more of microorganisms than we are of ourselves. It’s an incredible thing! So you can understand why a pneumonia virus can knock a person down. There are trillions of individuals producing something toxic.

I became aware of your work in the early 1990’s, thanks to your research on ethanol. One of your projects, having to do with the application of microorganisms in the production of alcohol, had yielded a magic patent number in the United States, 5 million, which was, incidentally, also front page news in the Gazeta Mercantil at that time.
I’ll try to summarize this story. Two months after I finished my doctorate in 1971 at USP, I went to the University of Bath in England, to do a postdoc. I went there to work with the physiology of yeast and had chosen to study Saccharomyces cerevisiae. At university, it is interesting to think about how we end up becoming an expert in some matter. In fact, I went to Bath to study the membrane of S. cerevisiae, or rather, the fatty acids of membrane phospholipids – the unsaturated ones. With this knowledge, I later worked with the action mechanism of antibiotics active in the yeast membrane. In 1975 the “Pro-Alcohol Program” was initiated in Brazil, which used S. cerevisiae to produce alcohol. I became interested, I thought it was a great opportunity to employ my sound knowledge of this yeast. I went to visit alcohol production plants to see how fermentation was being done, because it is one thing in the university where we work in volumes of 50, 100, 200 milliliters, and quite another in a factory using 300 or 400 thousand liter fermentation tanks. It is a vastly different scale and the problems involved are also different. I acquired a great deal of knowledge, became involved with professors at Esalq [Luiz de Queiroz Agricultural College], from there I worked as a consultant for some time at IPT [São Paulo Institute for Technological Research] in the area of alcohol fermentation, and I was working on this when I thought that I should really go to a more advanced research center. I met Professor Lonnie Ingram, of the University of Florida, and I thought, “Wow, this guy does really cool work on alcohol fermentation.” I applied for a grant from CNPq [the National Council for Scientific and Technological Development in Brazil], to go and work with Prof. Ingram and it was approved. I arrived at his laboratory, began to work with alcohol. Soon I saw that he was starting a new project: using a genetically modified bacterium to produce ethanol using genes from Zymomonas mobilis. I got interested in this line of research and wrote a short letter to CNPq saying: “I would like to change my project because they are also starting a new project in the area of alcohol.” In fact, they were already working with a genetically modified organism. And I was lucky, so to speak … because of that patent … My whole life is full of luck.

Is luck not the result of hard work to some extent?
I have a lot of luck. Talking a gene from one organism and transferring it to another is easy to do. But this gene must then be expressed. Furthermore, the gene needs to remain stable within the new host organism and also have a long period of productivity. And it was precisely this that the Americans knew how to do. The group was very strong in terms of molecular biology, genetics, the technical aspects of transferring genetic material, it was just a good understanding of the physiology of the bacterium that was lacking. When the research trials began, we only managed to produce a very small amount of alcohol. This is where my luck in understanding alcohol fermentation and the bacteria itself, which does not normally ferment and produce alcohol, came into play. With the adaptations I was making in the process, the microorganism began to produce a sufficient quantity of alcohol, which ultimately became interesting from a technological standpoint.

What exactly was the knowledge that you contributed to this line of research in Florida?
The physiological processes of an organism are always dependent on multiple variables. You need to tune, to harmonize the information and behavior of the microorganism. You adjust the pH, mess with the temperature, add some substance that interferes with or slows the rate of growth. Adjust the concentration of a particular salt, or of a cation or an anion. These are the settings. And in this case, for an organism that has never produced alcohol. We simply wanted the organism to produce the greatest quantity of alcohol possible – and it turned that when we gave sugar to the organism, it was able to produce alcohol. That’s what we did and it was a great result. What is even more interesting though, is that we used some of the properties of Escherichia coli, which has the ability to use many different sugars. When I say use, I mean to grow and multiply. Normally, Saccharomyces cerevisiae can not ferment these sugars that exist in nature. With the genes that we introduced from our bacteria, the microorganism began to use sugars, and began producing ethanol from xylose.

And “our bacteria” is the very E. coli itself?
Yes, genetically modified E. coli.

To what extent was this knowledge added to the manufacturing process in Brazil and the United States?
Zero.

Why?
The patent came out in 1991. We have to think that countries were, and still are today, heavily dependent on petroleum. Dependent not only on fuel, but also dependent from the standpoint of the entire industrial complex. And alcohol could have competed with gasoline. At that time, the oil and gas industry was very powerful, as it is today, and did not want to see alcohol being used in any country. Even here in Brazil, at that time, the price of ethanol was not competitive with that of gasoline, so everyone thought that alcohol did not need to be produced. So much so that in the late 1990’s, Brazil nearly terminated the Pro-Alcohol Program.

But after oil prices increased …
Increased again, then alcohol became more interesting. But no one even talked about pollution, sustainability.

EDUARDO CESARAnyhow, the ethanol issue began to turn into something more desirable and increased interest in getting ethanol from cellulose, the new generation of ethanol, a course already followed for five years …
It is the pursuit of technologies for ethanol production on a new base. To produce ethanol, you cut the sugarcane, take it to the production plant, remove the juice, and ferment it to produce ethanol. What remains is known as bagasse. And the bagasse still has residual cellulose, hemicellulose and lignin. Cellulose and hemicellulose are sugar polymers, potentially convertible into alcohol by some kind of organism, if we are able to do this. You would have to break down the cellulose. For the hemicellulose, our process was adequate. And when I returned from the United States, ran to numerous plants in the interior of São Paulo, saying, “Look, there is a process that allows the hydrolysis of sugarcane bagasse. From this hydrolysis you can make alcohol. This will produce about 10% to 15% more alcohol if we develop the technological processes”. There in the United States we did this on a small scale. These two raw materials are already in the factory. To be able to transform all of this into ethanol is everyone’s dream. Only nobody has succeeded yet.

And why not?
Because the chemical breakdown of cellulose is difficult to do. From an industrial point of view, it’s not possible. You can break down cellulose, there are enzymes for that, but it is not yet economically viable. What I find interesting in this whole story, is that at the end of last year I went to Campinas, to CTBE [National Laboratory of Bioethanol Science and Technology]. I went to see what was being proposed, second and third generation ethanol, and so on. And I wasn’t surprised when the researchers there told me the following: “We’ve invited you here because we want to talk a little bit about E. coli.” I showed them a study that had come out at the end of last year, demonstrating that the best known organism for producing ethanol so far is E. coli. They were trying to resuscitate it. My process was developed 17 years ago. It’s an old process.

What is your expectation regarding the technologies that are being developed right now?
When I was still working as a researcher on this subject I thought that the question would be resolved very quickly from the technological point of view. Because you simply have to do something called enzymatic hydrolysis, or chemical hydrolysis. My experience is with chemical hydrolysis. And I managed to do it, there are several graduate students who have done work with chemical hydrolysis, followed by fermentation. We have achieved the most promising results yet, like those of the Americans. It is a matter of having enough money to do it on a larger scale and seeing if they can get the same results. In microbiology there are no secrets. If you want to go to scale, you have do tests. It is not a chemical reaction, in which you just put a pound or 10 pounds or 100 pounds, and it will always be the same. It’s not like this with microbiology. One must invest in research and this is something that factories here in Brazil don’t like to do. When I was making my round of visits to sugar cane processing plants, I would try to generate interest in my ideas with them and the question was always “how much more money will we make?”. Never “how much will be need to invest?”.

What is the most advanced work in relation to the question of ethanol going on today?
I think that the cleanest way to do this will be when we have enough enzymes to breakdown all of these macromolecules. From that point on, we won’t need to use genetically modified microorgansims anymore. We will use the old and classic yeast method of producing alcohol from glucose.

Are we still a long way from achieving this?
From an industrial point of view, we are long way away.

What is the biggest challenge from an industrial point of view and from the standpoint of our degree of knowledge to achieve the total transformation of biomass into sugar … into glucose?
I believe that we have adequate knowledge of cell metabolism. Microorganisms have control over their own metabolic activity. Ourselves and microorganisms, because, biochemically, you are just like a bacterium. If the cells in our body are in the presence of a sugar, for example, glucose in the presence of another sugar, xylose, the cell will use the glucose first, then go to another sugar, then to a third sugar, then to a fourth sugar. The organism has a sort of saving mechanism. It already has an organized metabolic pathway. When you take such a biomass, in which there are many different sugars together, the microorganism will first have to use one of these sugars in order to then be able to use a different sugar. The biomass is still very large, with a complex chemical mix, and it won’t be possible to achieve this.

At what time in the future should this become possible?
Research depends on people and money. And alcohol is very interesting because we are highly dependent on oil prices. I’m not only talking about in Brazil but worldwide. Economically and culturally as well. I’m a little bit skeptical when I hear people say “No, but alcohol is less polluting than gasoline.” Sure it is! But price does not enter into this account. That’s coming, but much too slowly. This is why I still think that if the price of oil is high, great. Great because alcohol will be important. If oil prices go down (and Brazil has prospects of seeing the price of oil go down due to pre-salt), this will affect the use of alcohol.

Do you consider your work with Escherichia coli to be the peak of your entire research career? Or an aggregate to other studies?
I was always very much split between my teaching and research activity. I was always very dedicated to teaching, I’ve always been a teacher. And I think that it is to be both.

How so? University culture demands that you combine research, teaching and extension.
That’s right. It turns out that universities tend to place the greatest value on faculty doing good research, while good teachers go mostly unnoticed. I managed to survive being a teacher. And I like it.. I do it because I believe that the main function of a university is to pass knowledge on to students. The university has a responsibility to train people.

But is it not desirable to train people while also doing research?
You have courses in areas where you do not do research and achieve excellence in training people. Okay, research helps a lot when a person combines this sort of practical experience with knowledge. But like I said, I think it’s important to be a good teacher. This takes time. Each time you repeat a class, you have to think about it, what worked, what didn’t work, and what you can modify.

How did your teaching work and your research contribute to the book Microbiology?
I think that it also has a little bit to do with the idea of divulging the subject of microbiology, about access to microbiology. This textbook was written primarily for students in the field of medicine, particularly undergraduate and graduate level students. I’m always trying to spread information about microbiology.

Where does your fascination with microbiology come from?
From people that I met at USP. On the first day of classes, my microbiology professor at the Pharmacy and Biochemistry Faculty called on me and said: “I know you.” It scared me a little bit. He took out a photograph of my mother with a baby on her lap. “Your mother was my German teacher.” This professor, whose name was Lúcio de Carvalho Lima, was a wonderful person in my life. He died a long time ago. He was the one who motivated me. After I finished the course, he invited me to be his teaching assistant the following two years. He was completing his application to be a full professor. And I helped him with this work.

What was the subject of the thesis?
The production of pyocyanin, which is a pigment from Pseudomonas aeruginosa. After I finished college in 1964, he invited me to be his assistant. At that time there was no testing required in order to be accepted into higher education, it was by recommendation, he was a top professor. I learned a lot about being a teacher from him. He was an excellent teacher.

Returning to your book, some editions were done in partnership with the microbiologist, Luiz Rachid Trabulsi. In fact, he started the book project alone, right?
Trabulsi had already completed two editions. He started when he was at the Medical School at USP, then he went to the São Paulo School of Medicine. Some time later, when I was head of the department at USP, he joined the department. I already had a strong connection with him because I was very involved with the Brazilian Society for Microbiology. I served as the treasurer, secretary, president, and vice-president. After many years I was the editor of the Journal of Microbiology (Revista de Microbiologia) while he was president of the society. After the second edition of the textbook, he invited me to be a co-editor. We completed the third and fourth editions together. But by the time the fifth edition was in progress, he had passed away and I did it alone.

You’ve gone from writing a technical manual to creating a story for children. How do you foresee the next few years of your productivity?
I’m working on another book to fill this gap. I reached the end of graduate school to the initial phase of education. There is an interface here that I want to fill. But this book has stayed on standby, so to speak. The impact of the children’s book was amazing to me. In just a month after being launched, the book was nearly sold out already. 750 copies were printed. I was responsible, together with Telma, for this print run, did you know that?. We are going to make a book about sustainability and there are people asking me to do another book about sexually transmitted diseases. These are ideas that are just appearing. I don’t want to do it alone though. I want to see if I can get people who have this ability, of being able to put stories on paper in a way that is enjoyable, easy to understand, and based on accurate knowledge.

If you had to summarize your career, would you say that there is a stage when you focused more on research and then came back to teaching?
My departure from the university occurred for the following reason. In 2003 there was a possibility that an American firm, which had acquired the rights to patent number 5 million, would set up its operations here in Brazil. They had been talking with an ethanol production plant in Goiás where they wanted to initiate a new production process using our bacteria. I was really excited about this possibility. To get involved with it, I had to leave the university. I had already completed many years of service, and in October I requested permission to retire, because in January I expected to start working with the private firm. The president of the company that had bought the firm came to Brazil in December, he called me and said, “Look, Brazil is no longer our priority, we will not do any more business here. None of our previous plans are to be pursued.” And there I was, retired. And I had stopped doing research at the university, I left the university at that particular moment, because I was going to be able to conduct the best kind of research, which was to install my process in an alcohol production plant.

The American company had no strategic vision.
But do you know who the owner of this new firm is? He is a former mayor of the city of St. Louis and was a figurehead of the oil industry. He wanted to destroy the company that developed our process at the University of Florida.

And what was that company?
It’s called Quadrex. It never even really got started. It was all settled in the United States when Bio Energy, which was the original firm, was purchased by Quadrex.

So you mean that your difficult experience had nothing to do with the university?
No. It was the private industry that frustrated me at this time, an American company that was going to do a project in Brazil that would have been the pinnacle for me in terms of pursuing work that I had started in the laboratory and taking it to an industrial scale. I was cut off at that moment and I missed the bus.

But did you later receive any resources for the licensing of patent number 5 million?
No, because it was not yet licensed, since the company gave up. The company had purchased a license to use the patent. But the patent was still held by the University of Florida, until it was bought by a Japanese plant that was producing alcohol from sawdust using our bacteria. Sawdust is also the same thing (as sugarcane), cellulose and hemicellulose. When the patent was about to expire, I received a letter from the University of Florida saying: we are going to pay you royalties . They sent me $340. I was really happy because I had earned that $340. What I find interesting is that all of this brought me satisfaction. This is the funny side of the story. Because it’s obvious that I had never planned on making money. It’s interesting that the news about the patent came out in the New York Times. It was then that O Estado de S. Paulo, reading the New York Times, learned of the existence of a Flavio Alterthum. The news came out here in Brazil and I was interviewed on the Jô Soares show, Fantástico, Jornal Nacional, and on CNN. People like to be on the crest of a good wave. In fact, I am happy.

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