Team:Alberta-North-RBI E/projectsolution

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== Our Microscopic Friend ==
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==Our Solution==
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As bio-processing technologies continue to improve, the use of biomass as a renewable resource will become more and more attractive. Biomass exists in many forms and is produced by diverse industrial processes. Paper recycling plants, in particular, generate a copious amount of waste biomass in the form of paper sludge. Costly disposal of this waste stream typically occurs via landfill, or through inefficient combustion. This is somewhat shocking because paper sludge is actually a significant source of valuable cellulose. Through our integrated bio-processing strategy, we can exploit this cellulose to produce high-value chemicals while simultaneously having a positive impact on waste management.
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At '''Upcycled Aromatics''', work is underway on the design of an integrated bioprocessing facility. In conjunction with an enzymatic hydrolysis, engineered bacteria will transform the paper sludge waste stream into aromatic platform chemicals. The engineered bacteria will have the ability to produce a variety of aromatic chemicals - controlled by process conditions such that modification of one parameter results in the production of one specific chemical. The biosynthetic pathway under investigation will allow us to produce shikimic acid, cinnamic acid and it's derivatives.
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We intend to engineer a ''Pseudomonas putida'' strain containing a “switch” system enabling the manufacture of either shikimic acid (SA) or cinnamic acid (CA) as a major product. Future embodiments of this strain will contain additional directing mechanisms whereby a multitude of derivative chemicals can be favored.
 
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The enzymatic activities leading to the production of SA will be enhanced by the addition of extra-chromosomal copies of endogenous genes modified such that expression is constitutive. Where necessary, these genes will be further modified through site-directed mutagenesis to abolish feedback regulation. Optimization of metabolism to SA is based on the invention(s) described in US patents 5168056 and 6613552.
 
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Enzymatic activities which lead from SA to phenylalanine (Phe) will be abolished by removing the underlying endogenous genes. These endogenous genes, in addition to a codon-optimized ''Rhodosporidium toruloides'' gene encoding for a phenylalanine ammonia-lysase (PAL) enzyme, will be re-introduced at an extra-chromosomal locus as modified genes containing inducible promoter/repressor “switches” allowing us to direct metabolism toward CA at will. The “switch” mechanism will require “cis” DNA elements immediately upstream of each enzyme’s coding sequence (CDS). In addition, each regulatory system (ie. induction, repression) will require a constitutively-regulated “trans” element encoded at a different locus which produces the protein component.
 
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To enhance the yield of our desired products, two additional categories of changes will be made to our host strain. Enzymatic activities which direct metabolites away from the desired pathway will be abolished by removing the underlying endogenous genes and transporter proteins responsible for re-uptake of SA or CA from the medium will be removed in the same manner.
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<p>The details of our solution can be found in two sections: </p>
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Latest revision as of 20:32, 26 October 2012

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Our Solution

As bio-processing technologies continue to improve, the use of biomass as a renewable resource will become more and more attractive. Biomass exists in many forms and is produced by diverse industrial processes. Paper recycling plants, in particular, generate a copious amount of waste biomass in the form of paper sludge. Costly disposal of this waste stream typically occurs via landfill, or through inefficient combustion. This is somewhat shocking because paper sludge is actually a significant source of valuable cellulose. Through our integrated bio-processing strategy, we can exploit this cellulose to produce high-value chemicals while simultaneously having a positive impact on waste management.


At Upcycled Aromatics, work is underway on the design of an integrated bioprocessing facility. In conjunction with an enzymatic hydrolysis, engineered bacteria will transform the paper sludge waste stream into aromatic platform chemicals. The engineered bacteria will have the ability to produce a variety of aromatic chemicals - controlled by process conditions such that modification of one parameter results in the production of one specific chemical. The biosynthetic pathway under investigation will allow us to produce shikimic acid, cinnamic acid and it's derivatives.



The details of our solution can be found in two sections:



















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