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-----Original Message-----
From: Biotech-Mod3
Sent: 12 December 2008 11:15
To: 'biotech-room3@mailserv.fao.org'
Subject: 75: Re: Biogas production - potential, large plants, education program

This is Emma Kreuger from Sweden again.

In the background document, "second generation biofuels" refers to liquid fuels from lignocellulose. My question is 'Do the fuels need to be liquid': This question is not directly focused on biotechnology but considering the amount of research efforts put into biotechnology to produce liquid fuels (e.g. enzyme production) it is important to consider what we are focusing on and why, as commented earlier by Dr. Ruzena Svedelius.

Also, responding to message 74 by Uwe Bruenjes:

Methane (the energy carrier in biogas and natural gas) can, contrary to what is claimed in the message, be used for cars, trains, ships and plains directly (there are examples of all) or via electricity. [In discussing biogas, Section 3.3.b of the background document reads: "The resulting fuel can be used for heat, electricity and as a vehicle fuel (after the gas has been compressed and using the same engine and vehicle configuration as natural gas).". For more on this subject, see e.g. Borjesson, P. and B. Mattiasson. 2008. Biogas as a resource-efficient vehicle fuel. Trends in Biotechnology, 26: 7-13, or http://www.eurobserv-er.org/pdf/baro186_a.pdf (July 2008, EU biogas barometer, 2 MB)...Moderator].

For countries that do not have a well developed infrastructure, it is definitely worthwhile to think about what would be the best infrastructure. Gas grids instead of electricity grids, cars that run on gas and homes that produce electricity? Or, are electrical vehicles the way to go? Should everything possible be converted to electricity; biogas, biomass, sun, wind etc. Imagine the air in Mexico city if all cars ran on electricity. Imagine how little energy it takes to transport gas in a pipeline.

Anaerobic digestion does not need to be made by "hobby gadgets" just because it is possible. It can be very energy efficient on an industrial scale. What we convert the biomass to, and how we do it, I do not care about. What is important is the total resource efficiency and the environmental effects.

Emma Kreuger,
PhD student
Department of Biotechnology
Lund University
Sweden
emma.kreuger (at) biotek.lu.se
+46 222 81 93
http://www.biotek.lu.se/research/renewable_energy/

-----Original Message-----
From: Biotech-Mod3
Sent: 12 December 2008 11:18
To: 'biotech-room3@mailserv.fao.org'
Subject: 76: Biochemical conversion of LC biomass - GM corn

This is Professor J. Ralph Blanchfield, an independent food scientist and Immediate Past President of the International Academy of Food Science and Technology.

The following may not be on the immediate application agenda, but I am surprised that no contributor so far has pointed out that there is a very well-known biochemical converter of lignocellulosic (LC) biomass, namely the cow. This fact prompted research at Michigan State University (MSU), the results of which were announced in April this year and presented by Mariam Sticklen, MSU professor of crop and soil science, at the 235th national American Chemical Society meeting in New Orleans and later in an article "Plant genetic engineering for biofuel production: Towards affordable cellulosic ethanol" in the June edition of Nature Review Genetics, 9: pages 433-443. [The article's abstract reads: "Biofuels provide a potential route to avoiding the global political instability and environmental issues that arise from reliance on petroleum. Currently, most biofuel is in the form of ethanol generated from starch or sugar, but this can meet only a limited fraction of global fuel requirements. Conversion of cellulosic biomass, which is both abundant and renewable, is a promising alternative. However, the cellulases and pretreatment processes involved are very expensive. Genetically engineering plants to produce cellulases and hemicellulases, and to reduce the need for pretreatment processes through lignin modification, are promising paths to solving this problem, together with other strategies, such as increasing plant polysaccharide content and overall biomass" (http://www.nature.com/nrg/journal/v9/n6/abs/nrg2336.html) ...Moderator].

Quoting from the MSU news story released about this in April 2008 (http://www.news.msu.edu/story/872):

"The enzyme that allows a cow to digest grasses and other plant fibers can be used to turn other plant fibers into simple sugars"..."MSU scientists have discovered a way to grow corn plants that contain this enzyme. They have inserted a gene from a bacterium that lives in a cow's stomach into a corn plant. Now, the sugars locked up in the plant's leaves and stalk can be converted into usable sugar without expensive synthetic chemicals"... "As reported, turning plant fibers into sugar requires three enzymes. The new variety of corn created for biofuel production, called Spartan Corn III, builds on Sticklen's earlier corn versions by containing all three necessary enzymes.

The first version, released in 2007, cuts the cellulose into large pieces with an enzyme that came from a microbe that lives in hot spring water. Spartan Corn II, with a gene from a naturally occurring fungus, takes the large cellulose pieces created by the first enzyme and breaks them into sugar pairs. Spartan Corn III, with the gene from a microbe in a cow, produces an enzyme that separates pairs of sugar molecules into simple sugars. These single sugars are readily fermentable into ethanol, meaning that when the cellulose is in simple sugars, it can be fermented to make ethanol. "It will save money in ethanol production," Sticklen said. "Without it they can't convert the waste into ethanol without buying enzymes - which is expensive."

The Spartan Corn line was created by inserting an animal stomach microbe gene into a plant cell. The DNA assembly of the animal stomach microbe required heavy modification in the lab to make it work well in the corn cells. Sticklen compared the process to adding a single Christmas tree light to a tree covered in lights. "You have a lot of wiring, switches and even zoning," Sticklen said. "There are a lot of changes. We have to increase production levels and even put it in the right place in the cell." If the cell produced the enzyme in the wrong place, then the plant cell would not be able to function, and, instead, it would digest itself. That is why Sticklen found a specific place to insert the enzyme. One of the targets for the enzyme produced in Spartan Corn III is a special part of the plant cell, called the vacuole. The vacuole is a safe place to store the enzyme until the plant is harvested. The enzyme will collect in the vacuole with other cellular waste products. Because it is only in the vacuole of the green tissues of plant cells, the enzyme is only produced in the leaves and stalks of the plant, not in the seeds, roots or the pollen. It is only active when it is being used for biofuels because of being stored in the vacuole."

Prof J Ralph Blanchfield, MBE
Food Science, Food Technology and Food Law Consultant
Fellow, Institute of Food Technologists
Fellow and Immediate Past President (2006-08), International Academy of Food Science and Technology
United Kingdom
Personal Web address: www.jralphb.co.uk
e-mail: jralphb (at) easynet.co.uk

[While on the issue of genetically modifying corn to produce enzymes for biofuel purposes, something recent and related, although dealing with first-generation rather than second-generation biofuels, that might be of interest is that in a news release of 24 November 2008, the United States Department of Agriculture's Animal and Plant Health Inspection Service (APHIS) said it was seeking public comment on a petition to deregulate corn genetically engineered to produce a microbial enzyme that facilitates ethanol production. In the United States, there are two types of ethanol processing plants, dry-grind (accounting for ca. 80% of ethanol production) and wet-milling plants (ca. 20% of ethanol). The corn variety (Event 3272) is being produced for dry-grind ethanol production and it contains the amy797E gene encoding the heat-stable AMY797E alpha-amylase enzyme. Alpha-amylase is used to break down the starch component of the corn into dextrins, maltose and glucose and the product concept is that the Event 3272 grain will serve as the source of amylase enzyme in the dry-grind ethanol process, replacing the addition of microbially produced enzyme. See the news release and documention about Event 3272 corn at http://www.aphis.usda.gov/newsroom/content/2008/11/deregcorn.shtml and http://www.regulations.gov/fdmspublic/component/main?main=DocketDetail&d=APHIS-2007-0016 respectively...Moderator].

-----Original Message-----
From: Biotech-Mod3
Sent: 12 December 2008 11:31
To: 'biotech-room3@mailserv.fao.org'
Subject: 77: Re: Biogas production - potential, large plants, education program

This is Emma Kreuger again, responding to message 73 by Anju Arora about heating of reactors for biogas production:

To keep an anaerobic reactor warm during winter time (and at a stable temperature all year) it needs to be insulated. For a cooperative, I believe it should be possible to finance such investments and also to finance electricity generators. Income can be generated by producing and selling electricity (for vehicles or other purposes) or by selling gas as vehicle fuel. Heat for heating the reactor can be collected from the sun or part of the heat produced together with electricity can be used to heat the reactor.

Anaerobic digestion of cellulosic biomass at lower temperatures has been studied in our group, but it was found to be very slow at temperatures below 25 C (Bohn I, Bjornsson L and Mattiasson B 2007, 2 articles). Some studies indicate that 55 C would be superior to 37 C for cellulose degradation (manure contains much cellulose). By using insulation and heat exchange it doesn´t cost much more energy to run the process at 55 C than 37 C.

Emma Kreuger,
PhD student
Department of Biotechnology
Lund University
Sweden
emma.kreuger (at) biotek.lu.se
+46 222 81 93
http://www.biotek.lu.se/research/renewable_energy/

[The two Bohn et al 2007 articles mentioned are probably: i) The energy balance in farm scale anaerobic digestion of crop residues at 11-37 C. Process Biochemistry, 42: 57-64 and ii) Effect of temperature decrease on the microbial population and process performance of a mesophilic anaerobic bioreactor. Environmental Technology, 28: 943-952...Moderator].

-----Original Message-----
From: Biotech-Mod3
Sent: 12 December 2008 15:33
To: 'biotech-room3@mailserv.fao.org'
Subject: 78: Re: Biotechnology applications and Jatropha curcas

This is from Dr. K. Chalapathy Reddy, Mission Biofuels India Pvt, again

Indian scenario - In problems we find opportunities

All developing nations should work towards becoming 'energy independent' which is one of the important areas to upgrade them to become developed nations in the near future. Among different types of energy sources, bioenergy through plant- and/or animal-based feedstock has to play a great role to reduce the dependence on non-renewable energy. Plant-based feedstock for bioenergy is one among energy sources which is very promising because of its renewable nature and sustained production with low cost maintenance. Plants yielding oil are considered suitable for production of biodiesel. Since there is huge requirement of edible oils for daily consumption, the best alternate option is to go for non-edible oil yielding plants for Indian conditions. In this context, Indian planners, scientists and industries are focusing on non-edible oil yielding plants as a source of ideal feedstock for producing biodiesel. Among plants; Tree Borne Oil Seeds (TBOs) are found more suitable for biodiesel production and among these are Pongamia (Pongamia pinnata), Neem (Azadirachta indica), Jatropha curcas and Mahua (Madhuca longifolia). Each one of these plants has advantages and disadvantages. However, Jatropha curcas has been projected by and large. This plant has the potential and for sure will find itself in the drivers seat as biodiesel feedstock in coming days; provided the following areas needs to be addressed together.

Jatropha curcas: A potential feedstock for biodiesel

Since it is an introduced species to India long back, so this species has a narrow genetic base. However, many have reported the existence of large phenotypic variation across India and this has been attributed to the influence of local environmental factors. There was a Jatropha Network Project initiated by the Government and it is a collaborative project between institutes/universities/organizations. This project has given lot of details on Jatropha germplasm available and the variability existing for agro-economically important traits within India. For example Indian Jatropha curcas material has the variability for few traits like oil content ranges from 27 to 54 percent, test weight ranges from 1250 to 1720 seeds per kilogram; branching angle, sex ratio, fruiting behavior, tolerance to drought and frost. After this, very few institute(s) started continuing the further work on Jatropha independently and few collaborations with industries; we have started creating variability through intraspecific and interspecific crosses. Earlier it was reported one sucessful interspecific cross but now we have observed seed setting in three interspecific crosses. This has resulted in creating more variability for many traits. There is huge scope for creating variability beyond imagination in Jatropha through conventional breeding program.

Conventional and molecular marker breeding

Currently, efforts are focusing on molecular characterization of the existing and developed breeding materials for developing molecular markers. Progenies obtained from interspecific crosses were involved in back crossing with Jatropha curcas. This resulted in developing more variability for agro-economically important traits like female:male flower ratio, fruit size, seed weight and plant type. With every backcross generation, there was improvement in the traits of our interest. These populations were subjected for molecular characterization for developing markers; however this is in initial stage.

Like this few institutes and companies are working in different direction with a strong genetic base but lack facilities for biotechnological tools to combine with conventional breeding program to jumpstart genetic improvement for seed and oil yield characteristics of Jatropha curcas. On the other side, few institutes/organizations have started developing molecular makers for important traits since they have infrastructure but they do not have access to the diversified germplasm materials. So these developments are happening independently across India and other nations. So for quick improvement in Jatropha seed and oil yield we should come forward for working together to reduce the gaps between the institutes/organizations. Otherwise, after two years also we will be debating the same issues of low genetic variability and low yields of Jatropha; so Jatropha curcas is not-feasible as a feedstock for biodiesel production. We should also understand the intellectual property rights (IPR) issues related to sharing the knowledge, technology, breeding materials and/or the IPR generated during the collaborations.

Dr. K. Chalapathy Reddy
Senior Scientist
Mission Biofuels India Pvt Ltd,
608, 6th Floor, Powai Plaza,
Hiranandani Business Park,
Powai, Mumbai- 400076
India
Tel: +91-22-25706216, +91-22-32601001
Telefax +91-22-25706215,
Mob:+91-9323623328
Url : www.missionnewenergy.com
E-mail: dr.chalapathy (at) missionnewenergy.com

-----Original Message-----
From: Biotech-Mod3
Sent: 12 December 2008 15:53
To: 'biotech-room3@mailserv.fao.org'
Subject: 79: Biogas and biotechnology

This is Joy Clancy again.

I wanted to respond to the discussion on biogas and biotechnology. I wrote a paper for the Dutch Ministry of Development Cooperation (DGIS) on the potential contribution of biotechnology for biogas production in developing countries in 1994. One of the recommendations was that priority should be given to biotechnological research which increases the range of feedstocks, through pre-treatment of biomass, and the development of specialised inocula for hostile environments (for example, in cold mountainous regions where biogas production can drop drastically or virtually cease in the winter months). This could contribute to a reduction in digester cost and an increase in the number of people who have access to the technology by extending the resource base. Nevertheless, it has to be concluded that the major constraint on increased access to the technology is primarily a socio-economic one, related to income levels in rural areas, and not one of lack of access to a good technology.

I don't think that any of my recommendations were taken up and I still think the report is valid!

Joy Clancy
Dept of Technology and Sustainable Development (TSD)
Centre for Clean Technology and Environmental Policy (CSTM)
The University of Twente
PO Box 217, 7500 AE Enschede,
The Netherlands
email: j.s.clancy (at) utwente.nl
telephone: +31-53-4893537(direct);3545(sec)
fax: +31-53-4894850
web site: http://www.utwente.nl/cstm/tsd/
Urban energy: http://www.urbanenergy.utwente.nl/


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