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COMPANY RESEARCH

It’s time for biotechnology in biomass

GranBio invests in R&D to meet the challenges of second generation ethanol

Pereira, standing at left, and the GranBio research and development team in Campinas. In the rear, bales of straw from sugarcane

Léo Ramos Pereira, standing at left, and the GranBio research and development team in Campinas. In the rear, bales of straw from sugarcaneLéo Ramos

The straw remaining in the field after sugarcane is harvested is the principal raw material GranBio uses in ethanol production. In September 2014, the company became the first in Brazil to produce cellulosic ethanol, or second-generation (2G) ethanol made from biomass by a biotechnological process on an industrial scale. Traditionally. First generation ethanol fuel was produced from sugarcane juice. Headquartered in the capital city of the state of São Paulo, this industrial biotechnology company was founded in 2011 and operates over the entire length of the supply chain. GranBio also pursues scientific and technological solutions on several fronts of the agricultural and industrial production system.

Company
GranBio
R&D Center
Campinas, São Paulo State
Nº of Employees
18
Main products
Development of technology for ethanol production from the leaves and stalks of sugarcane

The company has already made important technological strides, such as a genetically modified yeast and a variety of sugarcane aimed at producing 2G ethanol. The Bioflex 1 plant was set up in São Miguel dos Campos, in the northeastern state of Alagoas, next to the Caeté Plant, which produces first-generation ethanol. GranBio obtains sugarcane straw from this and three other plants in the region. Cellulosic ethanol is produced experimentally in only a few commercial plants around the world. There are two in the United States, which uses the stems and leaves of maize as a raw material; one in Italy, which uses wheat leaves; and in July 2015, the Brazilian energy company Raízen opened a plant using sugarcane bagasse and straw in the city of Piracicaba, São Paulo State.

GranBio’s initial strategy was to import technology from abroad to speed up the production process. A number of ingredients were chosen, such as yeasts from the Dutch company DSM and enzymes from the Danish company Novozymes. At the same time, the company set up a research and development (R&D) center in Campinas. “What we do is convert science into technology,” says Gonçalo Amarante Guimarães Pereira, a GranBio partner and chief scientist of the company. At age 51, Pereira has been a professor for 18 years at the Genomic and Expression Laboratory of the Institute of Biology of the University of Campinas (Unicamp). Most of the members of the research team he leads were recruited from the university. Currently there are 18 researchers and technicians working directly on R & D: eight with doctoral degrees, two with master’s degrees and four doctoral students.

In Pauliceia (São Paulo State)

GranBio In Pauliceia (São Paulo State)GranBio

Special yeast
The team was responsible for two recently announced technological achievements. The first is the development of a yeast capable of processing xylose, the sugar present in hemicellulose, one of the three main fibers of sugarcane leaves and stems, along with cellulose and lignin. “In the first generation, the industrial yeast strain (of the Saccharomyces cerevisiae species) consumes the sucrose and fructose found in soluble form in sugarcane juice to produce ethanol; in bagasse, the sugars in the fibers of the leaves, such as xylose and pentose, are not soluble and so the yeast does not recognize them,” says Pereira. To make yeast capable of processing xylose, the GranBio team developed a genetically modified strain with a gene from another microorganism—which they prefer not to reveal—and some modified genes of the Saccharomyces species itself.

The organism was approved for commercial use by the general coordination office of the National Biosecurity Commission (CTNBio) in April, and its use is the subject of a patent filed with the Brazilian Industrial Property Institute (INPI). “We will start using genetically modified yeast on the production line in 2016.” The use of xylose, according to Pereira, represents a profit of approximately R$50 million annually for the company, which intends to process 400,000 metric tons of biomass per year. “Glucose makes up 40% of this material, pentose approximately 35%, and xylose 25%, which translates into about 100 metric tons. For the second generation to be profitable, xylose and other sugars found in the straw and bagasse must be processed.”

Sugarcane straw in the molecular biology laboratory

GranBioSugarcane straw in the molecular biology laboratoryGranBio

The genetic engineering work done on yeast was led by biologist Leandro Vieira dos Santos, 32. “We did a study to identify genes and combinations of genes that would induce Saccharomyces to consume xylose,” says Santos, who is working on a doctorate at Unicamp. A graduate of the Federal University of Viçosa (UFV), where he received a master’s degree in microbiology, Santos, after working two years at biotechnology company Agrogenética, decided to do a doctorate focusing on yeast and contacted Pereira in 2011, when the researcher was putting together the GranBio team. Today, with the yeast ready, Santos is engaged in perfecting it.

The microorganism’s propagation and time scale are the work of biotechnology engineer Luige Calderon. He is originally from Peru, where he graduated from the Catholic University of Santa Maria in Arequipa, obtained a master’s degree from Unicamp and, in 2012, was recruited to work at GranBio as a researcher in the bioprocess area. “I select microorganisms using genetic and evolutionary engineering, and then develop, for example, the most appropriate means of cultivation,” Calderon explains. Yeast genetics also drew Osmar de Carvalho Netto to the company. He has a degree in food science from the University of São Paulo (USP), and a doctorate from Unicamp. He participated in the sequencing of the Saccharomyces genome at Unicamp and thought about forming an industrial yeast company with Santos. “We were going to be one of GranBio’s suppliers, but they convinced us to join the company instead,” he says. And so he has become the company’s process coordinator for areas such as fermentation, hydrolysis and pretreatment of biomass. He was also assigned to bridge the gap between research and the corporate area. “It had to be someone who understands the language of scientists and I now spend half my time on this. The other half is spent on process coordination,” he says. For example, he is involved in organizing tests, deliveries of materials and staying in touch with the plant. All this is done so that GranBio’s researchers can concentrate on research.

Sugarcane samples in the analytical chemistry laboratory

GranBioSugarcane samples in the analytical chemistry laboratoryGranBio

Back to the beginning
In addition to yeast, another GranBio innovation was the energy-sugarcane introduced in August. It is a new non-transgenic variety developed with traditional crosses between several other cultivars in collaboration with the University Network for the Development of the Sugar-Energy Sector (Ridesa) of Alagoas and the Campinas Institute of Agronomy (IAC). “We went back to the beginning of breeding sugarcane. Instead of increasing sugar in the juice, we boosted the amount of fiber in the plant. And so we have a more rustic sugarcane. It is taller, has harvest longevity, flowers less and is more disease-resistant as well as being harder,” says the agronomist José Bressiani, the company’s agricultural director. With undergraduate, master’s and doctoral degrees in breeding from USP, he gained experience in producing sugarcane cultivars during his 15 years working at the Sugarcane Technology Center (CTC) plus five years at Canavialis, part of the Monsanto group. “My job is to think about the biomass function for sugarcane plants. We are creating a plant that should have low production costs,” he says.

Ethanol test in biochemical analyzer

GranBioEthanol test in biochemical analyzerGranBio

The tests with energy-sugarcane, which bears the trade name Cana Vertix, are taking place in the Southeast, Northeast and Central-West. The idea is that, in the future, specific sugarcane varieties will be used only to produce 2G ethanol and to generate electric energy by burning waste. Initially, only the straw of Cana Vertix will be used. After mechanical harvesting of the sugarcane, the straw remains behind on top of the soil and dries after a few days, after which it is collected and transported to the 2G plant, or it can be stockpiled for several months.

Technological advances and the growth of GranBio increased the importance of the R & D center, which is now a subsidiary known as BioCelere. The company’s formation began with a conversation between Pereira, Bernardo Gradin, president, and Alan Hiltner, executive vice president of GranBio, in March 2010. Gradin was stepping down as president of Braskem and wanted to invest in biotechnology and 2G ethanol. “We were at a restaurant and on the back of the check I outlined what the future GranBio could do in the scientific field. He liked it and then invited me to be one of the partners,” says Pereira. The two had met before—they served together in the Army in Bahia and were reunited in 2007 when Pereira coordinated a project for the Partnership for Technological Innovation (PITE) of FAPESP, between Unicamp and Braskem, on renewable propylene (raw material for the production of plastics) made with sugarcane.

070-073_GranBio_235-02GranBio’s investment has already reached $265 million for a production capacity of 82 million liters per year. As of August 2015, production had reached a total of 3 million liters of ethanol. It was not high as we had hoped, because some industrial processes needed improvement. “But we have already found solutions and are going to implement them in early 2016,” says Pereira.

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