New Breakthrough Biotechnology Adds Value to Corn Growers Output & Reduces Costs of Ethanol Biofuel.

A new cheaper way of liquifying corn starch for biofuel production is making its way to the market from Swiss biotechnology company Syngenta (Syngenta AG (ADR) Public, NYSE:SYT)

Syngenta have developed a catalyst works in water that's hotter even than boiling water.

This a real step forward for renewable fuels technology that will improve the economics of greenhouse gas emission control via biofuels, and add value at the farm-gate to corn grains used for fuels.

Syngenta has developed their heat-resistant amylase starch liquification catalyst (AMY797E) to be expressed in corn Line 3272 grain crops that are destined for dry-grind fuel ethanol production in the United States.

The fact that the catalyst for starch processing comes from bacteria that grow in water that is hotter than boiling, being under natural high pressures in the deep ocean, shows that these are truly amazing catalysts. A rich record of scientific research as been done on them, as illustrated by publications noted at the end of this post.

In the near future, this new corn (maize) will most likely be used widely in the USA to reduce the costs of making ethanol biofuels.

The by-products of this biofuel process - such as distillers solubles - are very suitable for the animal feed markets. For this reason, food use regulatory approval for the feed is needed in countries to which the USA might export the corn, and this is now being pursued world-wide by Syngenta.

Already in South Africa, the potential competitive edge provided by this new corn to American corn growers has been recognised.

Microbial enzymes already are widely used in the starch processing industry already, often as Genetically Engineered versions of natural catalysts.

In this case a new version of the microbial enzyme amylase has been developed that is active when exposed to heat needed to ready corn starch for ethanol production. It comes as a hybrid enzyme generated from three closely related Archaea bacteria (Thermococcales, Thermococcus), long familiar to microbiologists because their relatives make methane. The bacteria (more correctly Archaea) in this case come from the sea. This is very innovative and exciting modern biotechnology.

Not only that, Syngenta's approaches cuts out manufacturing steps in amylase production by getting the corn plants to make their own catalyst amylase out in the corn fields. This is a truly green and clean approach.

Microbially produced alpha-amylases are already commonly used commercially in the starch-processing step during corn dry-grind and wet milling processing. The purpose of the development of Line 3272 corn, as it is called, is to use the corn grain as the source of amylase enzyme in the dry-grind ethanol production, replacing presently added of microbially produced enzyme, and reducing costs.

For links to other recent crop technology breakthroughs, and more details on heat-tolerant amylases click this hyperlink.

See also
Diversa Corporation News:

Amylase-T

Corn Amylase Improves the Efficiency of Ethanol Production

Syngenta is developing a genetically modified strain of corn that expresses high levels of alpha amylase—a thermal-tolerant digestive enzyme developed by Diversa that turns the corn’s starch into sugar for ethanol.

The engineered plants are designed to reduce costs by eliminating the need for mills to add liquid enzymes. The Amylase-T seeds do not increase the yield, rather they make corn easier to process which translates into substantial savings for mill operators.

Syngenta has announced that pilot trials have been successfully conducted and that it anticipates launch of this product in 2008.

Main Story Ends Here, Technical Details follow:

An application for regulatory approval of this corn, related to its value as an animal feed, has already been submitted to Australian Food Safety Agency FSANZ.

FSANZ. INITIAL ASSESSMENT REPORT, APPLICATION A580
FOOD DERIVED FROM AMYLASE-MODIFIED CORN LINE 3272

Line 3272 falls within the scope of the US Food and Drug Administration’s (FDA) 1992 Statement of Policy: Foods Derived from New Plant Varieties, including genetically engineered varieties pursuant to 21 CFR Section 192.25 of the Federal Food, Drug, and Cosmetic Act. Syngenta has initiated a consultation with FDA and filed a Pre-market Biotechnology Notification (PBN) in September 2005.

A Petition for the Determination of Nonregulated Status for Corn Line 3272 was submitted to the USDA in October 2005.

During 2006, dossiers for food and/or feed approvals will be submitted to the relevant authorities in the European Union, Canada, China, Japan, Korea, Philippines, Taiwan, Russia, South Africa and Switzerland.

Technical details:

Event 3272 is a genetically modified (GM) maize that expresses a synthetic amy797E gene encoding the thermostable AMY797E alpha-amylase protein. Alpha-amylases are key additional components in the production of ethanol derived from maize. These enzymes catalyse hydrolysis of starch into smaller and less complex carbohydrate molecules during the starch liquefaction step of the dry-grind ethanol process. The expression of AMY797E is specifically targeted to the grain endosperm and the genetic modification has been engineered to retain the protein in the endoplasmic reticulum of the cells. Maize grain from Event 3272 expressing the AMY797E alpha-amylase enzyme will serve as the source of amylase enzyme in the dry-grind ethanol process, replacing the external addition of microbially produced enzyme. Event 3272 also expresses the PMI protein that allows transformed maize cells to utilize mannose as a sole carbon source and acts as a selectable marker.

The amy797E gene, a chimeric [hybrid] gene derived from three wild-type alpha-amylase genes from the archael order Thermococcales. The gene encodes a thermostable alpha-amylase protein, selected for its increased thermostability and activity during high temperatures required for starch hydrolysis in dry-grind ethanol production

The most relevant published scientific papers follow. Clearly many others are treading the same path as Syngenta:

Landry TD, Chew L, Davis JW, Frawley N, Foley HH, Stelman SJ, Thomas J, Wolt
J, Hanselman DS. Safety evaluation of an alpha-amylase enzyme preparation derived from the archaeal order Thermococcales as expressed in Pseudomonas fluorescens biovar I. Regul Toxicol Pharmacol. 2003 Feb;37(1):149-68.
The Dow Chemical Company, Midland, MI 48674, USA.
[Pundit note: Probably this is not related to Syngenta project directly]

BD5088 alpha-amylase derived from archaeal sources has characteristics of pH and temperature tolerance that are well suited to hydrolysis of starch in food processing applications. The production microorganism recipient strain, Pseudomonas fluorescens biovar I, strain MB101, was avirulent after oral administration to mice and does not represent an infectious threat to humans.
Repeated dose gavage studies with BD5088 enzyme preparation, up to 13 weeks in duration, showed no systemic toxicity due to the oral route with an NOAEL of 890 mg/kg/day as Total Organic Solids. Some irritation occurred in the respiratory tract, which was considered to be a consequence of reflux and aspiration of test material that contained lipopolysaccharide from the Pseudomonas production strain. A 2-week dietary study (0 and 310 mg/kg/day) confirmed that there were no respiratory tract effects related to oral ingestion. There was no genotoxic activity based on Ames, mouse lymphoma, mouse micronucleus, and rat lymphocyte chromosomal aberration tests. There was no evidence of allergenic potential based on a comparison of the primary sequence of BD5088 with sequences in an allergen database. The enzyme was labile to pepsin digestion. Based on these data, BD5088 alpha-amylase preparation may be considered safe for use in food production such as corn wet milling.

Leveque E, Haye B, Belarbi A.
Cloning and expression of an alpha-amylase encoding gene from the hyperthermophilic archaebacterium Thermococcus hydrothermalis and biochemical characterisation of the recombinant enzyme. FEMS Microbiol Lett. 2000 May 1;186(1):67-71.

Dickmanns A, Ballschmiter M, Liebl W, Ficner R.
Structure of the novel alpha-amylase AmyC from Thermotoga maritima. Acta Crystallogr D Biol Crystallogr. 2006 Mar;62(Pt 3):262-70. Epub 2006 Feb 22.

Yang SJ, Lee HS, Park CS, Kim YR, Moon TW, Park KH.
Enzymatic analysis of an amylolytic enzyme from the hyperthermophilic archaeon Pyrococcus furiosus reveals its novel catalytic properties as both an alpha-amylase and a cyclodextrin-hydrolyzing enzyme. Appl Environ Microbiol. 2004 Oct;70(10):5988-95.


Grzybowska B, Szweda P, Synowiecki J.
Cloning of the thermostable alpha-amylase gene from Pyrococcus woesei in Escherichia coli: isolation and some properties of the enzyme. Mol Biotechnol. 2004 Feb;26(2):101-10.

Berk H, Lebbink RJ.
High-throughput screening of mutant alpha-amylase libraries for increased activity at 129 degrees C. Methods Mol Biol. 2003;230:127-35. No abstract available.

Savchenko A, Vieille C, Kang S, Zeikus JG.
Pyrococcus furiosus alpha-amylase is stabilized by calcium and zinc. Biochemistry. 2002 May 14;41(19):6193-201.

Savchenko A, Vieille C, Zeikus JG.
alpha-Amylases and amylopullulanase from Pyrococcus furiosus. Methods Enzymol. 2001;330:354-63. No abstract available.

Linden A, Niehaus F, Antranikian G.
Single-step purification of a recombinant thermostable alpha-amylase after
solubilization of the enzyme from insoluble aggregates. J Chromatogr B Biomed Sci Appl. 2000 Jan 14;737(1-2):253-9.

Jones RA, Jermiin LS, Easteal S, Patel BK, Beacham IR.
Amylase and 16S rRNA genes from a hyperthermophilic archaebacterium. J Appl Microbiol. 1999 Jan;86(1):93-107.

Janecek S.
Sequence of archaeal Methanococcus jannaschii alpha-amylase contains features of families 13 and 57 of glycosyl hydrolases: a trace of their common ancestor? Folia Microbiol (Praha). 1998;43(2):123-8.