Mesas Redondas

 

Roundtable 1: Industrial Production of 2G Ethanol in Brazil 

 

606

COORDINATOR: Carlos Alberto Labate (CTBE)

[email protected]


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648

PANELIST: Carlos Eduardo Vaz Rossell (ctbe)

[email protected]

 

Industrial Production of Ethanol 2G: Critical Points

 

In this opportunity critical points to be considered for the design of a 2G ethanol process will be discussed. Initially, a short presentation of the 1G ethanol process will be delivered in order to point how it can contribute for 2G ethanol production. Following, P&D and Engineering requirements of the 2G ethanol process will be commented, concerning:

Pretreatmeant processes;

Enzymatic Hydrolysis: development and formulation of the enzymatic complex; modeling and design of the reactor;

Recovering of pentoses and their further processing to ethanol;

Doubts related to the use of GMO in Industrial bioprocesses at large scale;

After, it will be commented the present situation of the projects related to the Industrial Program of 2G ethanol: pretreatment, enzyme production, enzymatic conversion, etc. Finally, it will be presented the state of the laboratories and pilot plant of CTBE, and how they can contribute to the scientific community that is researching the conversion of lignocellulosic material.

 

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 612

PANELIST: Célia Maria Araújo Galvão (CTC)

[email protected]

 

CTC Technology for Second Generation Ethanol Production (CTC-E2G):

Challenges and Perspectives

 

Sugarcane biomass (bagasse and straw) has been pointed out as a great alternative to fossil resources for the production of fuels. It presents all the requirements for utilization as raw material to supplement the ethanol domestic production, without needing any expansion of the planted area, and clearly increase the productivity of this biofuel per hectare, making the Brazilian bioethanol even more competitive. The technology developed by CTC for the production of second generation ethanol was based on the extensive knowledge that CTC has on the sugarcane industry and is designed to conform to the Brazilian reality, with specific solutions for the utilization of bagasse and straw, besides being fully integrated, in terms of energy and infrastructure, to the current industrial ethanol production – the so-called first generation. This feature is undoubtedly one of the main competitive advantages of CTC-E2G Technology, which is based on the continued development in seven years of research on different scales, from the laboratory to tests with continuous processing of thirty tons of sugarcane biomass. This approach allowed identifying the main bottlenecks of the process and also the most promising technological route and with greater economic viability. The final challenge of the CTC-E2G Technology before its release to the market, scheduled for 2016, is to validate the technical process parameters and consolidate the engineering in order to demonstrate its viability in semi-industrial scale. The expected positive results in the short-medium term will certainly help to secure the leadership position that Brazil now occupies in the biofuels world scenario.

 

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618

 PANELIST: Osmar V. Carvalho-Netto (GRANBIO)

[email protected]


First Industrial Cellulosic Ethanol Plant in the Southern Hemisphere


The increasing cost of oil and concern about the decline of its offer in the future, associated with environmental concerns, have motivated the search for alternative strategies for producing fuels and biochemicals from renewable sources. This source replacement is an enormous challenge that can only be overcome by countries with competitive features in climate, land availability, soil quality and technology. GranBio was born of the vision to bring about a green revolution capable of transforming the real potential of Brazilian biomass into energy abundance. In the last year, the first commercial scale cellulosic ethanol plant in the southern hemisphere was announced by GranBio. The plant will have a nominal production capacity of 82 million liters of ethanol and the operation is expected to start in Alagoas, Brazil, at beginning of 2014. Initially, the production will use sugarcane straw as feedstock and the plant will work in cooperation with the first generation mills, presenting significant synergies to both sides. The company goes beyond producing ethanol. Acting from start to finish in the chain of production, and integrating proprietary technologies with those of the global leaders, GranBio offers cellulose-rich plant species, land use solutions, integrated industrial processes, enzymes and microorganisms adapted for industrial production and biochemicals. Here, we present the state-of-the-art of the technology as well the plans to use energy cane as feedstock for future cellulosic ethanol plants.


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Roundtable 2: Advances in Industrial Biotechnology for Pharmaceuticals and Cosmetics


651

  COORDINATOR: Maria Helena Andrade Santana (UNICAMP)

[email protected]

 

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  690

 PANELIST: Fabiana Munhoz (L’Oréal)

[email protected]


Biotechnologies as sustainable transformations for innovative cosmetic ingredients

 

How the development of biotechnology is transforming the cosmetic industry from today and tomorrow? It allows to design more sustainable and green processes, lowering their environmental impacts, and also to develop innovative ingredients and active, with improved performance and value. Different ingredients can be originated from biotechnology: a surfactant with anti-inflammatory or antimicrobial activities, a new pigment, an active ingredient for skin hydration, a new lipid for hair care or protection: just to name a few. In this presentation, I will show some examples of innovative products developed by L'Oreal that became possible thanks to the development of biotechnology.

 

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 693

 PANELIST: Marcos C. Alegria (CRISTALIA)

[email protected]

 

Biopharmaceuticals research, development and production at the Cristália laboratory

 

The Cristália Laboratory, established since 1972, was initially engaged in psychiatry field, followed by anesthesia and pain treatment. An R&D center was then created in Itapira (São Paulo State), in the mid-eighties, with the aim of establishing the capacity for a wide spectrum of research on health-related issues. Remarkable achievements of this center include the development of medications with improved efficiency or safety, most of which were patented. More than twenty five years after the release of the first human recombinant therapeutic proteins, a new class of pharmaceutical products has attracted the attention of pharmaceutical companies: the biosimilars. We can define these biopharmaceuticals as drugs obtained through biotechnological processes which are capable of retaining the key properties of reference standards with expired patents. The Brazilian production of biopharmaceuticals raises the possibility of achieving national independence in both the public and private sectors. This could also lead to price reduction and the promotion of local industry. It was within this scenario that the Cristália Laboratory, has, since 2005, been investing in the creation of an R&D capacity in the Biotechnology sector with the aim of generating biosimilars. Most particularly, Cristália built a cGMP Biotechnology Production Plant which contains two segregated areas: one for biopharmaceuticals using microorganisms as the expression system and another based in mammalian cells platforms. In 2012 a joint venture between Biolab, Eurofarma and Cristália created Orygen Biotecnologia, a new Brazilian biotechnology company formed as a direct result of a stimulus by the federal government to establish a capacity to manufacture biologics in Brazil. Orygen Biotecnologia will initially focus on the development of biosimilar monoclonal antibodies (Mabs) to supply the demands of the Brazilian Ministry of Health and its unified national health system (SUS), working in collaboration with public laboratories to achieve this aim.

 

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636

 PANELIST: Maria Antonieta Ferrara (FIOCRUZ)

[email protected]

 

Yeast asparaginase: Research and development of a therapeutic enzyme for leukemia treatment

 

The main drug used for the treatment of acute lymphoblastic leukemia is the bacterial enzyme asparaginase, which is available in Brazil as an imported medicine. The high cost and the external dependence supply of such an important drug create an uncomfortable scenario, including recurrent shortage problems. Despite the drug efficacy, the bacterial enzyme causes severe immunological reactions. The asparaginase II of the eukaryote Saccharomyces cerevisiae, coded by the ASP3 gene, is a potential candidate as a drug per se and for patients who have developed hypersensitivity to the bacterial asparaginase. Within this context, Farmanguinhos/FIOCRUZ and the Chemistry Institute/UFRJ have established a partnership aiming at the development of a yeast asparaginase antileukemic drug. As outcomes of the research carried out to date, the following steps have been successfully accomplished: (i) cloning and expression of the ASP3 gene in the methylotrophic yeast Pichia pastoris, under the control of the AOX1 gene promoter (in collaboration with the Department of Cellular Biology/UnB); (ii)  bioprocess for enzyme production at a 2-L bioreactor comprising a multi-stage fed-batch high cell density culture (in collaboration with the School of Chemistry/UFRJ); (iii) high-yield process for enzyme extraction from the yeast periplasm; (iv) protocol for enzyme purification at analytical scale (in collaboration with Oswaldo Cruz Institute/FIOCRUZ). These processes allowed the production of a high-purity asparaginase that is currently under structural and biochemical characterization. Subsequent efforts will focus on: (i) development of a scalable purification process; (ii) pre-clinical assays; (iii) development of enzyme formulation in order to increase the half-life and decrease the drug toxicity.

 

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642 

 PANELIST: Vânia Maria Pacchioni (CRODA)

[email protected]


CRODA: A Short Presentation


In this presentation we intend to show who is Croda, its markets of actuation, strategy and future beliefs. Why we believe Biotechnology is the best way looking for the sustainability. Examples of Croda products using white biotechnology (biotechnology applied to industrial processes), green biotechnology (biotechnology applied to plants and agriculture) and blue technology (biotechnology applied to marine and aquatic products). It will be also made an overview on the importance of biotechnology in the Croda history and the most recent installations to research and manufacturing biotechnological products.


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Roundtable 3: Challenges for the Brazilian production of enzymes for industrial applications

 

687  

 COORDINATION: Elba Pinto da Silva Bon

[email protected]

 

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654

PANELIST: Álvaro Batista Neto (Verdartis, UNESP)

[email protected]

 

Challenges for the Brazilian Industrial Enzyme Production

 

Nowadays, several industrial processes can use enzymes; however, a customization technology is necessary to produce enzymes with the desired characteristics. The Verdartis technology platform is able to adapt enzymes for a specific industrial process. The adaptation of enzymes for pulp bleaching at a process temperature above 80oC is an example of the application of the Verdartis technology. In this process, enzymes generations were derived from an enzyme with optimum activity at 55oC, changing its maximum activity for temperatures above 80oC. This enzyme provided the reduction of 18 to 25% of chlorine dioxide employed in the process of pulp bleaching pulp, without any increasing the effluent Biochemical Organic Demand (BOD) or impacts over the desired yield of the process. The good result obtained was related to the previous study of the enzyme wild characteristics in the beginning of evolution process. Molecular biology techniques for enzyme development and molecular simulation by bioinformatics methods were previously used to evaluate the enzyme characteristics. Besides adapting enzymes for the pulp bleaching operation, it was also necessary the development of new process for efficient engineered enzyme production. Regarding this aspect, a process development from Verdartis Company increased about 85 times the enzyme production as compared to the initial condition. The process developed by used proper culture media, optimized cultivation conditions for the microorganism that carried the genetic information of the evolved enzyme. The Verdartis technology was developed with governmental financial support from CNPQ, FAPESP and FINEP. Private groups partnership is not still accomplished due to the lack of tests in industrial scale which do not allow the industrial evaluation of thermostable enzyme application in the cellulose and pulp process.

 

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657

 PANELIST: José G.C.Pradella (CTBE)

Priscila S. Delabona, Daniel Kolling (CTBE), Cristiane S. Farinas (EMBRAPA)

[email protected]

 

Trichoderma harzianum and Penicillium echinulatum glucohydrolase process production development: from screening towards pilot plant

 

Second generation bioethanol production rests on the production of a cheap and effective cellulolytic cocktail to promote enzymatic hydrolysis of lignocellulosic material. Cellulolytic but also accessory enzyme activities have being demonstrated to be important to breakdown hemicelluloses and pectin plant cell wall components increasing cellulose accessibility to cellulases. We have being working with the isolation, screening and selection of biomass-degrading fungi species and analyzed the enzymatic complex produced by selected strains. Those data demonstrated that a new strain of Trichoderma harzianum P49P11 was a potential source of glycosyl hydrolases (GH) for lignocellulosic biomass deconstruction. Aiming the on-site GH production the influence of pretreated sugarcane bagasse (pSB), sucrose (SUC) and soybean flour (SF) was investigated in shaking flasks and controlled bioreactor experiments using experimental mixture design (EMD). It was found that pH 5.0 controlled with NH4OH aqueous solution and an optimized culture medium maximized Fpase biosynthesis rate up to 20 FPU/L h, 90.000 IU/L xylanase and 25.000 IU/L β-glucosidase activities using proper combination of pSCB, SF and SUC. Complementary analysis of T. harzianum secretome was performed to identify enzymes that could be missing in the enzymatic complex to achieve high-yield pSB hydrolysis. These analyses enabled the selection of pectinase and α-L-arabinofuranosidase (AF) to be further investigated as supplements to the crude enzymatic extract produced by T. harzianum. Penicillium echinulatum studies aimed to modulate cellulases and accessory enzymes hemicellulases and pectinase activities in using components culture media manipulation in shaking flasks and controlled bioreactor experiments with the aid of Design of Experiment (DOE). It has been shown that cellulase induction may be attained using steam pretreated sugar cane bagasse (pSB) (cellulose content of 60% w/w) and that industrial grade yeast extract (YE), soybean flour (SF) and wheat flour (WF) helped to modulate and significantly increase Fpase, xylanase, β-glucosidase and pectinase activities in the enzyme complex up to 5 FPU/ml, xylanase 250 IU/L. Moreover, mathematical modeling from DOE helped to tune the enzyme cocktail profile in terms of those enzymatic activities. In a hole it was possible to shape enzyme cocktails with these relevant enzymatic activities and it is believed that this strategy may be useful in plant cell wall deconstruction. The process was scale up and influence of agitation rate on GH rate production was assessed. Overall, it was demonstrated that T. harzianum P49P11 and P. echinulatum enzymes have a great potential to be used in the industrial deconstruction of biomass.

 

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660

 PANELIST: Rafael de Araújo Borges, Mauricio Sergio Esteller (PROZYN)

[email protected]

 

Enzymes Application in Industrial Processes

 

Nowadays at many industries, the use of biocatalysts has been used to replace chemical compounds that can be toxic and not safe to the environment. Also the biocatalysts are been used to develop new products that couldn’t be produced without the enzymes. Beverage, bakery, fuel ethanol, sucrose, chocolate, meat, gelatin, coffee, leather, paper and pulp, feed, dietary supplements, detergents, oil extraction, among others are some examples of common application of enzymes in industrial processes. Enzymes are produced by all live organisms. It accelerates the chemical reactions in selective ways as part of the metabolism. The enzymes act under mild conditions, what mean that it is ideal for use in food technology where the customer intend to modify selectivity food raw materials preserving nutrients properties. The industrial enzyme market size has been expanding in recent years with the improvement of R&D technology and the extension of application range. From 2001-2011, the global industrial enzyme market had grown of 6.8 percent, and in 2011 the global market valued at $3.5 billion, with a year-on-year growth rate of 6.1 percent. The Global Industrial Enzymes market is expected to grow at 7.49 percent over the period 2012-2016. One of the key factors contributing to this market growth is the increase in the end-user applications of industrial enzymes.

 

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Roundtable 4: Advances in Food Industrial Biotechnology

 

663

 COORDINATION: Francisco Maugeri Filho (UNICAMP)

[email protected]

 

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672

PANELIST: Carlos Ricardo Soccol (UPR)

[email protected]

 

Application of the biorefinery concept in the soybean industrialization and production of biochemical products

 

The global production of soybean, for the harvest of 2012/2013, is estimated as 285 million tons (USDA, 2013) and the Brazilian production will be 83 million tons, which represents around 30% of the global production. The major producers are the states of Mato Grosso do Sul and Paraná, contributing to 27 and 22% of the national production, respectively. Soybean cultivation in Brazil corresponds to a harvested area of 27,65 million hectares (CONAB,2013). The major part of the soybean produced and exported by Brazil is in the raw, non-industrialized form. The industrialization of soybean is extremely important for Brazilian economy because it would generate thousands of new employments. In this lecture we will present some results about an integrated process, in which the biorefinery concept was applied for integral exploitation of an industrial residue generated during manufacturing of Protein-concentrate of soybean and production of biochemical products. (bioethanol, xanthan gum, alpha-galactosidase and lactic acid).

 

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675

 PANELIST: Gláucia Pastore (UNICAMP)

[email protected]

 

 Transforming bioaromas

 

Aroma compounds influence greatly the flavor of food products and govern their acceptance by consumers and market success. The increasing consumer preference for natural products has encouraged remarkable efforts towards the development of biotechnological processes for the production of natural flavor compounds. Biotransformation of terpenes represents a very promising alternative for production of aromas, since allows overcoming the problems associated with chemical synthesis. The biological route implies low generation of toxic waste and high regio-and stereo-selectivity reactions. In addition, products obtained by microbiological methods can be labeled ´natural´ under Brazilian regulations.

 

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678 

 PANELIST:

Murillo Villela Filho (Evonik)

[email protected]

 

Biotechnology Beyond Technology - Transforming Ideas into Solutions

 

The growing world population raises the question how to nourish people in a world with limited resources. This question is particularly relevant regarding protein supply. With the worldwide enhancement of the standard of living, the demand for protein is constantly increasing. The supplementation of animal feed with limiting amino- acids enables a resource-efficient production of animal protein. Thus, methionine is produced from petrochemical feedstock to serve as a feed additive. An even higher overall amount of lysine is demanded for the supplement of poultry and swine nutrition. It is possible to produce lysine from renewable raw materials using a biotechnological process. However, the industrial production of lysine requires not only a suitable process, but also a specific set of conditions. This contribution describes how to find the conditions for the utilization of biotechnology in order to help the food production.  The existence of a technical process still requires the proper environment to be transformed into an industrial application. The biotechnological production of lysine with Corynebacterium glutamicum is an established process1. Nevertheless, its application requires a set of conditions leading to the attractiveness of the enterprise. Among the aspects to be regarded there are:

Feedstock: Abundant and affordable feedstock is a pre-condition for the operation of a process. Besides, the year-round availability and the price stability must be regarded.

 Energy: Sources must be available at the site of production.

 Equipment: Besides the sourcing of special equipment necessary for the high-tech process, the continuous operation must be warranted with maintenance and spare parts. This might be a challenge.

 Market for the final product: A production is only attractive if there is a market willing to reward the producer’s efforts. Thus it must be evaluated and quantified in advance. 

Supply chain & logistics: The raw materials must be available at the production site timely. In the same way, the product must be transported to the final customer with its desired properties. The supply chain concept is to be elaborated prior to start-up. 

Legal environment: The development of proprietary Intellectual Property (IP) is an integral part of an innovative and competitive industry, however resources consuming and expensive, Thus, its protection must be regarded prior to utilization.

Human resources: The availability of highly qualified and motivated personnel is essential for the operation and development of a high-tech process. This aspect must be carefully treated prior to and during production planning.

 

Based on the case study of Evonik Industries’ plans to implement a Biolys production facility in Brazil, all these aspects of biotechnology are described and commented. Only the combination of accretive conditions with the suitable process will enable biotechnology to deliver real solutions. Many of these conditions must be regarded at early stages of the process development. This case study calls the attention of the community for this essential aspect of their work, which is often neglected.

 

REFERENCES

1 Pfefferle, W.; Möckel, B.; Bathe, B.; Marx, A. Advances in biochemical engineering/biotechnology. 2003 59–112

 

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