Parker & Valentine-Cole -- Cannabis Compost US Patnet
Application

![](0logo.gif)  
**[rexresearch.com](../index.htm)**

---



**David PARKER & Virginia
VALENTINE-COLE**

**Cannabis Composter**




---



**US2009042974**   
**Compositions And Methods Relating To
Extensible Transgenic Vector Assembler, Pestilence Ridder,
Plus Cannabinoid Producer**

**Inventor(s):  PARKER ANSON DAVID [US]; VALENTINE-COLES
VIRGINIA LEE [US]**

**Abstract** -- The following provisional patent application
refers to the methods and compositions relating to a novel use
for enzymatic catalysis of C21-H30-O2 (delta-nine
tetrahydrocannabinol (THC)) as an insect repellent, bactericide,
and fungicide and dispensation methods as commercial reagent.
The bio-synthesis of cannabinoids represents a landmark
achievement in the field of composting, vector removal and
ecological reconstitution. Although the benefits have been known
for millennia, the advent of modern bio-engineering techniques
brings these small seeds of native wisdom to bear on a broader
and more industrialized scale-removing dangerous molds and
pestilences such as mosquitoes from swamped and flooded regions,
raw sewage areas, and disaster sites where ensuing vermin and
harmful vectors may cause greater damage than the initial
catastrophes.; It is the ambition and intention of authors that
tactical usage of this broad-sweeping technique may rapidly and
at low cost satisfy a global demand in what may be termed a
"grass-roots" bio-engineering project worthy of the 3rd
Millennium; bringing to fruition micro-mass-productivity. Should
a clear error be found either in spirit or in factual evidence
presented please do not hesitate to contact me.

**FIELD OF THE INVENTION**

[0001]The present invention relates the compositions and
methods for controlling harmful pestilence in toilets and
sewage, using modular extensible genetic techniques, and the
facilitation of rapid, low-cost, cannabinoid production.

**BACKGROUND OF THE INVENTION AND RELATED ARTS**

[0002]Regions of standing wastewater harboring high
concentrations of unprocessed, unfiltered rubbage and manure can
be natural sites for disease. The need for low-impact,
low-maintenance composting solutions is needed to address city
sewers and streets around the world that routinely overflow with
toxic bilge. Controlling pests in such regions may often require
the use of manufactured chemicals--created along costly and
inefficient energy gradients (USPTO U.S. Pat. No. 5,227,537).
Many of these chemical treatments present long-term hazards to
the environment in the forms of run-off contamination, and
build-up. Once introduced into a system these non-biodegradable
inorganic compounds may not easily be eradicated Although these
methods may be suitable for certain bilge water, especially in
treatment facilities where build-up and run-off are not
concerns, they do not address the needs of farmers dealing with
compost piles, nor sewage water running rampant through the
streets of cities that have been obliterated by tsunamis,
hurricanes, or tornadoes--wherein there may be an immediate need
for rapid decomposition and pestilence ridding. Nor do these
prior arts address the need for low impact conversion and or
transmutation of toxins and pestilence, nor do these methods
establish within the bilge an ecological breeding ground wherein
one skilled in the arts might hope to use the nutrient rich,
albeit highly toxic, solution for the establishment of
biological byproduct. On the other hand, allowing the
degradation of the toxic bilge to take place naturally may not
be a viable option--as the aforementioned pestilence may soon
make a bid to use the nutrient source as a home. In the proposed
invention one skilled in the arts would safely apply numerous
vectors to convert the bilge into valuable natural resources
while simultaneously defending the region from harmful
pestilence both through the creation of anti-microbial
substances and through direct competition for resources--in much
the same way that acidophilus in yogurt out--competes other
microbes.

[0003]With regards to the issue of treating filth dispersed
deep in underground sewers and inaccessible areas--the invention
makes a stark contrast to previous arts. Whereas most chemical
reactions must obey the laws of Brownian motion or undergo
energetically unfavorable processes such as pumping (USPTO U.S.
Pat. No. 5,360,556) or heating (USPTO U.S. Pat. No. 6,753,536),
enzymatic reactions enabled in motile vectors hold a decisive
advantage as they can move through a liquid medium more easily.
As one skilled in the arts appreciates the possibility of using
a motile plant vector such as the sperm of the gingko would
allow even greater motility for the vector. In the preferred
embodiment of this invention catalysts hosted in transgenic e.
coli, transgenic tobacco root hair, and used in modular
extensible vectors controlling the synthesis of compounds such
as tetrahydrocannabinolic acid (THCA), cannabigerolic acid
(CBGA), cannabichromenic acid (CBMA), the associated long term
costs of pestilence control may be reduced dramatically--while
simultaneously enriching the soil with valuable nutrients for
commercial crops. As one skilled in the arts will
appreciate--the long term application of the proposed invention
will manifest itself in stages--much as any great culture
ranging from ancient cheese and yogurt cultures to present day
bio-engineered vectors, each application of the invention may,
in the spirit of evolution, lead to a unique bio-transformation
specifically adapted to its environment. The proposed invention
brings to the table a base level of safer transmutation of
certain toxic fungi (Llewellyn 1977), (Turner 1981), infectious
microbes, (Van Klingeren 1976), (Schmitz 1973), and insect
pests, (Quaghebeur, 1981) as well as infectious disease
transmitted through insects such as West Nile Virus (McPartland,
1993). The nature of this invention is energetically favorable,
easily propagated, and low up-keep in cost making it also ideal
for third-world implementation in the pursuit of cleaner, safer
land. In cases of emergency the preferred embodiment might also
serve as a possible source of the neuroprotectant delta-nine
tetrahydrocannabinol (THC) through the application of heat such
as sunlight or direct flame. In the event of a terrorist attack
of neurotoxins, for instance, one might as a means of last
resort set fire to the growth medium to convert THCA to
THC--which upon inhaling provides neuroprotection (Hampson 1998)
(Van der Stelt 2001) (Mechoulam 2001) (USPTO U.S. Pat. No.
6,630,507). Whereas in prior arts Elsohy et al (USPTO U.S. Pat.
No. 6,730,519) disclosed a method for reduced cost THC
production they also rely on traditional abiotic, inorganic,
energetically unfavorable means for THC extraction and
purification of THC. Moreover their claims depend on natural
growth of Cannabis Sativa, a process that may take up to fifteen
weeks. Clearly this is not an acceptable waiting period in the
case of a terrorist attack. In an alternate embodiment of the
invention a serum of raw nutrients, as opposed to raw sewage,
were used as the basic medium--in this case using modularized
transgenic enzymatic techniques one skilled in the arts might
produce several tons of THC in two to three days.

**SUMMARY OF THE INVENTION--OBJECTS**

[0004]The term "Transgenic Stilbene-carboxylate synthase-like
enzyme (TSCSL)" (see Fellermeier 1998) refers to any enzymatic
reaction that yields Olivetolic Acid. The trigger mechanism. In
alternate embodiments of this invention it is linked operably to
a bioluminescent and equipped with a unique "off switch."

[0005]The term "Transgenic Geranylpyrophosphate Prenylase
(TOAP)" refers to any enzymatic reaction that yields
Cannabigerol (see Fellermeier 1998). In an alternate embodiment
linked operably to a bioluminescent and equipped with a unique
"off switch".

[0006]The term "Transgenic Cannabigerolic Acid Synthase (TCAs)"
(See Raharjo 2002) refers to any enzymatic reaction or nano-bot
that synthesizes Cannabigerolic Acid. In alternate embodiments
of this invention it is linked operably to a bioluminescent and
equipped with a unique "off switch."

[0007]The term "Transgenic Cannabidiolic Acid Synthase (TCBAs)"
(see Taura F. 1996) refers to any enzymatic reaction that
synthesizes Cannabidiolic Acid. In the preferred embodiment of
this invention it is linked operably to a bioluminescent and
equipped with a unique "off switch."

[0008]The term "Transgenic Tetrahydrocannabinolic Acid Synthase
(TTAs)" refers to any enzymatic reaction that synthesizes THCA,
(see reference Taura 2004). In an alternate embodiment of this
invention it is linked operably to a bioluminescent and equipped
with a unique "off switch."

[0009]The term "Transgenic Cannabichromene Synthase (TCBMs). In
an alternate embodiment of this invention it is linked operably
to a bioluminescent and equipped with a unique "off switch."
Such bioluminescent switch might include prior arts described in
USPTO U.S. Pat. No. 6,544,729, although one skilled in the arts
might determine others more suitable.

[0010]Genetic "Off switch"--any of several dozen enzymes with
known lethality targeting specifically the aforementioned
transgenic vectors--each with its own unique off switch.
Including but in no way limited to switches described in USPTO
U.S. Pat. No. 5,328,847.

[0011]The terms "wastewater, raw sewage, bilge water, manure,
compost, toxic sludge, filth, festering rot, crud, crude,
rubbage, and debris" refers to any medium that may need
pestilence management.

[0012]The term "pestilence management" refers to the
control--be it through repellence, extermination, or slowing of
growth rate, of any or several of the following organisms
Alabama argillacea (Riley 1885), Pieris brassicae (Beling 1932),
Melolontha melolontha (Mateeva 1995), and Aphelenchoides
composticola, (Grewal 1989), potato beetle (Leptinotarsa
decemlineata) (Stratii 1976), mosquito larvae (Anopheles and
Culex species)(Jalees et al. 1993), Chilo partellus, (a
lepidopteran borer)(Bajpai and Sharma, 1992), Tetranychus
urticae (Fenili and Pegazzano, 1974). Japanese beetles (Metzger
and Grant, 1932), Heterodera cajani (Mojumder et al. 1989),
Ustilago species (Misra and Dixit 1979, Singh and Pathak 1984),
Neovossia indica (Gupta and Singh 1983), Curvularia (Upandhyaya
and Gupta, 1989), Colletotrichum truncatum (Kaushal and Paul,
1989), Aspergillus, Penicillium, Cladosporium, Drechslera,
Fusarium, Cephalosporium, Rhizopus, Mucor and Curvularia
(Pandey, 1982), gram (+) S. aureus, Bacillus megaterium (Veliky
and Genest 1972), gram (+) Corynebacterium species and gram (-)
Pseudomonas and Agrobacterium species (Bel'tyukova 1962),
Trypanosoma brucei (Nok et al., 1994), Phomopsis ganjae (Charles
and Jenkins 1914, McPartland 1983), Arctia caja (Rothschild et
al., 1977) or any other known or unknown organism with
undesirable trails.

[0013]The term "transgenically enhanced vector" (TEV) refers to
any vector, its parental lineage or its offspring that has been
modified by the use of modern or Mendelian genetic techniques to
produce a compound.

[0014]The term "operably linked" refers to a juxtaposition
wherein the components so described are in a relationship
permitting them to function in their intended manner. A control
sequence "operably linked" to a coding sequence is ligated in
such a way that expression of the coding sequence is achieved
under conditions compatible with the control sequences.

[0015]Floatation system--in the preferred embodiment floating
systems with roots embedded are used to suspend the transgenic
roots as they convert cannabigerolic acid into cannabinoids.

[0016]The term "bioluminescent protein" refers to a protein
capable of causing the emission of light through the catalysis
of a chemical reaction. The term includes proteins that catalyze
bioluminescent or chemiluminescent reactions, such as those
causing the oxidation of luciferins. The term "bioluminescent
protein" includes not only bioluminescent proteins that occur
naturally, but also mutants that exhibit altered spectral or
physical properties.

[0017]The term "transformed" refers to a cell into which (or
into an ancestor of which) has been introduced, by means of
recombinant nucleic acid techniques, a heterologous nucleic acid
molecule.

[0018]The term "transgenic" is used to describe an organism
that includes exogenous genetic material within all of its
cells. The term includes any organism whose genome has been
altered by in vitro manipulation of the early embryo or
fertilized egg or by any transgenic technology to induce a
specific gene knockout.

[0019]The term "transgene" refers any piece of DNA which is
inserted by artifice into a cell, and becomes part of the genome
of the organism (i.e., either stably integrated or as a stable
extrachromosomal element) which develops from that cell. Such a
transgene may include a gene which is partly or entirely
heterologous (i.e., foreign) to the transgenic organism, or may
represent a gene homologous to an endogenous gene of the
organism. Included within this definition is a transgene created
by the providing of an RNA sequence that is transcribed into DNA
and then incorporated into the genome. The transgenes of the
invention include DNA sequences that encode the fluorescent or
bioluminescent protein that may be expressed in a transgenic
non-human animal, the genes required for the synthesis of
cannabinoids, and any additional genetic information necessary
for the greater control of the invention.

[0020]The following terms are used to describe the sequence
relationships between two or more polynucleotides: "reference
sequence", "comparison window", "sequence identity", "percentage
identical to a sequence", and "substantial identity". A
"reference sequence" is a defined sequence used as a basis for a
sequence comparison; a reference sequence may be a subset of a
larger sequence, for example, as a segment of a full-length cDNA
or gene sequence, or may comprise a complete cDNA or gene
sequence. Generally, a reference sequence is at least 20
nucleotides in length, frequently at least 25 nucleotides in
length, and often at least 50 nucleotides in length. Since two
polynucleotides may each (1) comprise a sequence (i.e., a
portion of the complete polynucleotide sequence) that is similar
between the two polynucleotides, and (2) may further comprise a
sequence that is divergent between the two polynucleotides,
sequence comparisons between two (or more) polynucleotides are
typically performed by comparing sequences of the two
polynucleotides over a "comparison window" to identify and
compare local regions of sequence similarity. A "comparison
window", as used herein, refers to a conceptual segment of at
least 20 contiguous nucleotide positions wherein a
polynucleotide sequence may be compared to a reference sequence
of at least 20 contiguous nucleotides and wherein the portion of
the polynucleotide sequence in the comparison window may
comprise additions or deletions (i.e., gaps) of 20 percent or
less as compared to the reference sequence (which does not
comprise additions or deletions) for optimal alignment of the
two sequences. Optimal alignment of sequences for aligning a
comparison window may be conducted by the local homology
algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2:482,
by the homology alignment algorithm of Needleman and Wunsch
(1970) J. Mol. Biol. 48:443, by the search for similarity method
of Pearson and Lipman (1988) Proc. Natl. Acad. Sci. (U.S.A.)
85:2444, by computerized implementations of these algorithms
(GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics
Software Package Release 7.0, Genetics Computer Group, 575
Science Dr., Madison, Wis.), or by inspection, and the best
alignment (i.e., resulting in the highest percentage of homology
over the comparison window) generated by the various methods is
selected. The term "sequence identity" means that two
polynucleotide sequences are identical (i.e., on a
nucleotide-by-nucleotide basis) over the window of comparison.
The term "percentage identical to a sequence" is calculated by
comparing two optimally aligned sequences over the window of
comparison, determining the number of positions at which the
identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs
in both sequences to yield the number of matched positions,
dividing the number of matched positions by the total number of
positions in the window of comparison (i.e., the window size),
and multiplying the result by 100 to yield the percentage of
sequence identity. The terms "substantial identity" as used
herein denotes a characteristic of a polynucleotide sequence,
wherein the polynucleotide comprises a sequence that has at
least 30 percent sequence identity, preferably at least 50 to 60
percent sequence identity, more usually at least 60 percent
sequence identity as compared to a reference sequence over a
comparison window of at least 20 nucleotide positions,
frequently over a window of at least 25-50 nucleotides, wherein
the percentage of sequence identity is calculated by comparing
the reference sequence to the polynucleotide sequence which may
include deletions or additions which total 20 percent or less of
the reference sequence over the window of comparison. As applied
to polypeptides, the term "substantial identity" means that two
peptide sequences, when optimally aligned, such as by the
programs GAP or BESTFIT using default gap weights, share at
least 30 percent sequence identity, preferably at least 40
percent sequence identity, more preferably at least 50 percent
sequence identity, and most preferably at least 60 percent
sequence identity. Preferably, residue positions which are not
identical differ by conservative amino acid substitutions.
Conservative amino acid substitutions refer to the
interchangeability of residues having similar side chains. For
example, a group of amino acids having aliphatic side chains is
glycine, alanine, valine, leucine, and isoleucine; a group of
amino acids having aliphatic-hydroxyl side chains is serine and
threonine; a group of amino acids having amide-containing side
chains is asparagine and glutamine; a group of amino acids
having aromatic side chains is phenylalanine, tyrosine, and
tryptophan; a group of amino acids having basic side chains is
lysine, arginine, and histidine; and a group of amino acids
having sulfur-containing side chains is cysteine and methionine.
Preferred conservative amino acids substitution groups are:
valine-leucine-isoleucine, phenylalanine-tyrosine,
lysine-arginine, alanine-valine, glutamic-aspartic, and
asparagine-glutamine.

[0021]Since the list of technical and scientific terms cannot
be all encompassing, any undefined terms shall be construed to
have the same meaning as is commonly understood by one of skill
in the art to which this invention belongs. Furthermore, the
singular forms "a", "an" and "the" include plural referents
unless the context clearly dictates otherwise. For example,
reference to a "restriction enzyme" or a "high fidelity enzyme"
may include mixtures of such enzymes and any other enzymes
fitting the stated criteria, or reference to the method includes
reference to one or more methods for obtaining cDNA sequences
which will be known to those skilled in the art or will become
known to them upon reading this specification.

**SUMMARY OF THE INVENTION--OPERATION**

[0022]As one skilled in the arts may appreciate the variety of
vectors able to transform of the initial reagents (TSCSL, TOAP,
TCAs, TTAs, TCBMs, TCBAs) into the desired reagents (THC, CBD,
CBM) may result in hundreds or thousands of potential scenarios.
Consider the heat that is generated in many compost conversion
where temperatures may rise above 160 degrees Fahrenheit, in
such cases it may be expedient to use an thermophilic vector,
particularly for the incubation of the TSCSL, and TOAP. In the
preferred embodiment of the invention it should be noted that
the TTAs, TCBMs, TCBAs, are used in either a plant or animal
vector--since cannabinoids exhibits both anti-microbial and
anti-fungal activity it will require a non-microbial and
non-fungal host.

[0023]In its preferred embodiment begin with a gigantic pile of
refuse, that may include fecal matter, untreated sewage water,
and decaying animal parts. It may be to the advantage of the
user to initiate the enzymatic activity in a more sterile
environment with nutrients needed for the synthesis of
precursors of cannabinoids to alleviate environmental pressures
of the sludge. In such cases as necessary the resulting enzymes
and precursor products may be added directly to the filthy
sludge or set aside and used as the growth medium for the
transgenic cannabinoid synthesis with the resulting cannabinoids
added to the filth sludge after their synthesis is completed.

[0024]Expose the pile of refuse to TSCSLs, TOAPs, and TCs
teas--brewed as per the guideline in the literature commonly as
anyone skilled in the arts will appreciate--and genetically
modified to include promoters operably linked to bioluminescent
proteins to help indicate and monitor effectiveness of the
treatment. This tea is given from 24 hours to one month as
indicated by bioluminescence (FIG. 1) to finish blending in with
the refuse--or as long as the bioluminescence appears active.
(FIG. 1. Step: Stilbene-like Synthase). These teas, when mixed
with toxic bilge, enzymatically synthesize cannabinoids.

[0025]Next a root bed made of TTAs, TCBAs, and or TCBMs. Note
the versatility of this invention. Any one of the aforementioned
synthases, or indeed all three may be placed atop the pile bilge
to create the desired reagents (ie THC, CBA, CBM). Also
noteworthy is the elegant closed-loop nature of this system. By
initiating the reaction with microbes that are not themselves
immune to the final product the system will eventually turn
itself off--as the reagent levels rise to higher levels the
TSCSLs, TOAPs, and TCs die.

**IN AN ALTERNATE EMBODIMENT**

[0026]The bucket containing the OAP is loosened atop a pile of
crud, that may consist of any decaying or decayed matter, and
that must consist of some decaying vegetable matter or living
vegetation.

[0027]The bucket containing the CAS is loosened atop the pile
of crud that previously received OAP treatment.

[0028]A blanket of roots from tobacco made of TTAs, TCBAs and
or TCBMs are thrown over the crapulence and festering therein
may it yield bountifully wee little cannabinoids.

**Overview**

[0029]This invention relates to the synthesis of cannabinoids
for the purpose of general pestilence riddance in filthy organic
and inorganic sludge. Through regulated enzymatic reactions,
wherein cannabinoids with known anti-microbial, insecticidal,
nematicidal, fungicidal properties and moreover nutritious, and
neuroprotective, qualities are used to benefit regions where
other commercial chemical reagents would require mechanized
dispersion and cleanup. In plain English for those skilled in
the arts--the genes involved in the enzymatic formulation of
cannabinoids are inserted into foreign vectors thereby
reproducing themselves and generating sufficient quantities of
cannabinoids to clear the region of pestilence.

[0030]The advantages of this system are numerous. Whereas
cannabinoid synthesis may not easily take place in Cannabis
sativa due to its illegality, this invention is highly
preferable. Whereas cannabinoid synthesis using inorganic
techniques is not advantageous due to the inefficiency of
inorganic and organic laboratory chemistry, this invention is
highly preferable. Whereas most chemical synthesis routes for
the creation of cannabinoids relate to the creation of extremely
pure cannabinoids, this invention merely creates sufficient
quantities as needed to rid a region of pestilence, and makes no
claims whatsoever as to purity. Whereas the cost of creating
cannabinoids synthetically would require large sums of money, as
well as recurring costs for reagents, as well as a high degree
of expertise and lab equipment, the invention described herein
requires a single up-front cost to create the necessary vectors,
and thereafter the invention may be distributed and applied to
sludge and filth across the world with almost no requirements
insofar a priori knowledge.

[0031]Using closed-loop modular enzymatic reactions, wherein
each phase of catalysis may be halted by another counter
reaction, and wherein each phase of catalysis may be easily
monitored for effectiveness allows one skilled in the arts to
more safely and effectively treat hazardous waste and the
plethora of contagions therein. This invention refers to
modular, in the sense that along the enzymatic pathway of choice
each enzymatic building block is separated into a unique vector,
uniquely identifiable by means bioluminescence and uniquely
susceptible to a flavor of anti-microbial or anti-fungal such
that the enzymatic process of choice may be halted at any given
phase of production if desired. Modular may also or rather refer
to the system as a whole, in that it should, handled by one
skilled in the arts, leave little or no trace of enzymatically
active reagent and be a closed-loop system--with the
understanding that in nature there exists no such thing as an
entirely closed-loop system, however, the preferred embodiment
of this invention has in its design constructs a self-destruct
or self-neutralizing mechanism for the living reagents. Thus in
the preferred embodiment of the invention the catalysis of
tetrahydrocannabinolic acid results in the recursive destruction
of the initial vectors (Taura 2004).

**DESCRIPTION**

[0032]Polyketide Synthesis converts 3 Malonyl CoA plus 1
n-Hexanoyl-CoA to form OSCoA. This conversion may take place
inside the muck and sludge, or may take place in a contained
area and after the OSCoA

[0033]Stilbene Carboxylate Synthase-Like (STCSL), in the case
of Cannabis Sativa a Chalcone Synthase (CHS) that exhibits
Stilbene Synthase (STS) activity in vivo and as per note in the
literature (Raharjo 2004) there is reason to believe that the
sequis used in the preferred embodiment of this invention and
refers to any enzyme that generates 5-amylresorcinolic acid
(olivetolic acid). While there are several enzymes capable of
synthesizing olivetolic acid in the final analysis any enzyme
capable of Olivetolic Acid synthesis will is sufficient. In the
preferred embodiment the STCSL is inserted into the
mitochondrial genome using the protofection technique (Khan
2004). The STCSL should be operably linked to bioluminescent
protein to facilitate the monitoring of activity. The vector of
the STCSL should also, in the preferred embodiment, have an
operably linked In an alternate embodiment of this invention
olivetolic acid is synthesized through inorganic techniques and
thus added to the filthy sludge as a trigger molecule. In this
manner the invention would have a limiting reagent from the
offset, restricting the final output of pestilence ridders in
such cases wherein limitations might be preferable. In another
alternate embodiment of the invention the STCSL is chimeric with
GOAP, or a pestilence ridding molecule.

[0034]Geranylpyrophosphate Olivetolic Acid Prenylase (GOAP)
(Fellermeier 1998) is an integral part of this invention, and
converts olivetolic acid into cannabigerolic acid. As one
skilled in the art may appreciate any enzyme capable of yielding
cannabigerolic acid is sufficient. In the preferred embodiment
the GOAP is loaded into the vector in the manner described in
Fellermeier's work. In the preferred embodiment of this
invention the GOAP is operably linked to a bioluminescent
protein such as GFP or aequorin, and thus its activation is more
easily monitored with minimal technical expertise. The GOAP is
also operably linked to a promoter capable of up-regulating GOAP
and thereby amplifying GOAP production.

[0035]Products made from these transgenic vectors should
produce THCA, and, in addition, other precursor molecules as
well as the necessary enzymes and proteins requisite for the
aforementioned production, such as, tetrahydrocannibigerolic
acid synthase, cannabigerolic acid synthase (CBGAS),
cannabidiolic acid synthase (CBDAS), cannabichromenic acid
synthase (CBRMAS), tetrahydrocannibinolic acid (THCA),
olivetolic acid, polyketide synthase, and cannabigerolic acid
synthase. Also disclosed is the unique and novel application of
the TTAs in the function of a compost toilet additive and for
the low-impact, sustainable, macrobiotic control of pests
including Alabama argillacea (Riley 1885), Pieris brassicae
(Beling 1932), Melolontha melolontha (Mateeva 1995), and
Aphelenchoides composticola, (Grewal 1989).

**Operation**

[0036]First the transgenically enhanced vectors (TEVs) as
necessary and leading up to the cannabigerolic acid phase of
biosynthesis (FIG. 1) are added into the growth medium and let
to rest for anywhere from 12 hours to several days with a
temperature range of 25-35 degrees centigrade, and also
depending on the volume of waste, the thickness of the muck, and
the general nature of the festering filth. If time is of the
essence one may speed up growth times by dispersing units of
TEVs around the afflicted region through artificial or assisted
means. If precision in timing is desired it may be convenient to
include a bioluminescent protein operably linked a functional
promoter to the TEVs similar in methods to (USPTO U.S. Pat. No.
6,544,729) and created such as to reflect the activity of the
TEVs.

[0037]Next the transgenic plant vector is placed atop the
festering sludge. The transgenic plant vector releases
cannabinoids into the sludge, and as it appropriates greater the
product of transgenic E. Coli(s) so shall it release
cannabinoids--all the while eradicating both the transgenic E.
Coli vector as well as the numerous pathogens, microbes,
insects, fungi etc . . . that are defenseless against the
cannabinoids.

**BRIEF DESCRIPTION OF THE DRAWINGS**

[0038]Many of the attendant advantages of the invention become
more readily apparent as the same become better understood by
reference to the following detailed description, which taken
with the accompanying drawing.

**FIG. 1 provides a basic visual understanding for one
skilled in the arts to process enzymatic THC, a drawing of the
3 phases of reaction.**

![](fig1.jpg)

![](fig2.jpg)

TABLE-US-00001 TABLE 1 Enzyme Reagents Product Time Temp Vector
Key References Accession # Polyketide 3 MalonylCo-A OSCoA 1-24
hrs 25-35 C. E. Coli M15 Raharjo, 2004 AY082343 STCSL/ & 1
n-Hexanoyl- Olivetolic Acid CHS CoA OSCoA CBDAs Cannabigerolic
Cannabichromenic 24-48 hrs 25-35 C. Tobacco Morimoto, 1999 Acid
Acid Root Hairs Prenylase Olivetolic Acid + GPP Cannabigerolic
Acid 1-24 hrs 25-35 C. E. Coli M15 Fellermeier, 1998 CBCAs
Cannabigerolic Cannabidiolic Acid 24-48 hrs 25-35 C. Tobacco
Morimoto, 1999 Acid Root Hairs THCAs Cannabigerolic
Tetrahydrocannabinolic 24-48 hrs 25-35 C. Tobacco Taura, 1995
& AB057805 Acid Acid Root Hairs Sirikantaramas 2004

**REFERENCES IN THE US PATENT OFFICE**

TABLE-US-00002 [0040]  
Author Title USPTO # Keyword Date   
Grobler, Marius; Sewage sludge treatment 20050175516 Compost NE
et al. Liang Shooting mechanism of an 6,615,815 Anti-violence
Sep. 9, anti-violence gun 2003 Becker, et al. Method and
arrangement of 5,692,446 Anti-violence Dec. 2, equipment for the
protection of 1997 buildings and people from acts of violence
Sun Apparatus for preventing 4,811,775 Anti-violence Mar. 14,
1989 criminal's escape or violence Peterson, et al. SPANN:
Sequence processing 5,067,095 Modular Vector Nov. 19, artificial
neural network 1991 Case, et al. Thin membrane sensor with
5,328,847 Modular switch Jul. 12, 1994 biochemical switch
Humphreys, et Apparatus for neutralizing 6,753,536 Wastewater
Jun. 22, 2004 al. chemical and biological threats cleaning
apparatus Ball, et al. Method of feeding wastewater 5,360,556
Wastewater Nov. 1, effluent to filter bed through cleaning 1994
parallel conduits apparatus Hampson, et al. Cannabinoids as
antioxidants 6,630,507 Cannabis Oct. 7, and neuroprotectants
Neuroprotectant 2003 Elsohly, et al. Method of preparing
delta-9- 6,730,519 THC synthesis Dec. 4, tetrahydrocannabinol
2001 Growcock, et al. Vermiculture compositions 6,838,082
compost Jan. 4, biolumin 2005 Sayler; Gary S. Bioluminescent
biosensor 6,544,729 Bioluminbiosensor Apr. 8, 2003 device device
Croteau, et al. Isolation and bacterial 6,258,602 cannabis Jul.
10, 2001 expression of a sesquiterpene insecticide synthase cDNA
clone from peppermint (mentha .times. piperita, L.) that
produces the aphid alarm phromone (E)-.beta.- farnesene Goodwin,
Neil Production of delta 9 20050171361 THC synthesis Aug. 4,
2005 John; et al tetrahydrocannabinol Martin, Billy R;
Cannabinoids 20050165259 Cannabinoids Jul. 28, 2005 et al.
Moore, Bob M. Cannabinoid derivatives, 20040242593 THC synthesis
Dec. 2, II; et al. methods of making, and use 2004 thereof
Chowdhury, Tetrahydrocannabinol 20040229939 THC Nov. 18, Dipak
K.; et compositions and methods of manufacture & 2004 al.
manufacture and use thereof use Webster, et al. Cannabinoid
extraction method 6,403,126 Cannabinoid Jun. 11, 2002 extraction
McKinney Method and apparatus for 4,279,824 THC extraction Jul.
21, 1981 processing herbaceous plant materials including the
plant cannabis

**REFERENCES IN THE LITERATURE**

[0041]1. Abe I, Watanabe T, Noguchi H. Enzymatic formation of
long-chain polyketide pyrones by plant type III polyketide
synthases. Phytochemistry. 2004 September; 65(17):2447-53.
[0042]2. Abrol B. K. and I. C. Chopra, 1963. Development of
indigenous vegetable insecticides and insect repellents.
Bulletin Jamu Regional Res. Lab 1:156. 1963.   
[0043]3. Bajpai N. K. and V. K. Sharma. Possible use of hemp
(Cannabis sativa L.) weeds in integrated control. Indian
Farmers' Digest 25(12):32, 38, 1992 [0044]4. Beling I.
Schadlingsbekampfung im 18. Jarhhundert. Anz.
Schadlingbekampfung 8(6):66-69., 1932.   
[0045]5. Business Alliance for Commerce in Hemp, "Hemp: Friend
to People and Ecology" Los Angeles, Calif., April 1994   
[0046]6. Cekmecelioglu D, Demirci A, Graves R E., Feedstock
optimization of in-vessel food waste composting systems for
inactivation of pathogenic microorganisms., J Food Prot. 2005
March; 68(3):589-96.  
 [0047]7. Chopra R. N., R. L. Badhwar and S. L. Nayar,
1941. Insecticidal and piscicidal plants of India. J. Bombay
Nat. Hist. Soc. 42:854-902.   
[0048]8. Dahiya M. S. and G. C. Jain, 1977. Inhibitory effects
of cannabidiol and tetrahydrocannabinol against some soil
inhabiting fungi. Indian Drugs 14(4):76-79.  
 [0049]9. Deportes I, Benoit-Guyod J L, Znirou D, Bouvier M
C., Microbial disinfection capacity of municipal solid waste
(MSW) composting., J Appl Microbiol. 1998 August; 85(2):238-46.
[0050]10. Eckermann C, Schroder G, Eckermann S, Strack D,
Schmidt J, Schneider B, Schroder J. Stilbenecarboxylate
biosynthesis: a new function in the family of chalcone
synthase-related proteins. Phytochemistry. 2003 February;
62(3):271-86.  
 [0051]11. Eisenreich, W., Schwarz, M., Cartayrade, A.,
Arigoni, D., Zenk, M. H. & Bacher, A. (1998) The
deoxyxylulose phosphate pathway of terpenoid biosynthesis in
plants and microorganisms. Chem. Biol. 5, R221''R233.   
[0052]12. Fellermeier M, Eisenreich W. Bacher A, Zenk M H.,
Biosynthesis of cannabinoids. Incorporation experiments with
(13)C-labeledglucoses. Eur J Biochem. 2001 March;
268(6):1596-604.   
[0053]13. Ferenczy L. Antibacterial substances in seeds. Nature
178:639-640., 1956.   
[0054]14. Ferenczy L., L. Gracza and I. Jakobey. An
antibacterial preparatum from hemp (Cannabis sativa).
Naturwissenschaften 45:188., 1958.   
[0055]15. Ferrer J L, Jez J M, Bowman M E, Dixon R A, Noel J P.,
Structure of chalcone synthase and the molecular basis of plant
polyketide biosynthesis. Nat Struct Biol. 1999 August;
6(8):775-84. Bioresour Technol. 2001 December; 80(3):217-25.   
[0056]16. Gal I. E., O. Vajda and I. Bekes. A kannabidiolsav
nehany tulaj-donsaganak vizsgalata elelmiszertartositasi
szempontbol. Elelmiszervizsgalati Kozlemenyek 4:208-216.1969.
[0057]17. Grainge M. and S. Ahmed. Handbook of Plants with
Pest-Control Properties. John Wiley and Sons, NY. 470 pp., 1988.  
 [0058]18. Grewal P. S. Effects of leaf-matter
incorporation on Aphelenchoides composticola (Nematoda),
mycofloral composition, mushroom compost quality and yield of
Agaricus bisporus. Annals Applied Biology 115:299-312., 1989.   
[0059]19. Gupta R. P. and A. Singh. Effect of certain plant
extracts and chemicals on teliospore germination of Neovossia
indica. Indian J. Mycology and Plant Pathology 13(1):116-117.
1983. [0060]20. Hampson A J, Grimaldi M, Axelrod J, Wink D.
Cannabidiol and (-)Delta9-tetrahydrocannabinol are
neuroprotective antioxidants. Proc Natl Acad Sci USA 1998 Jul.
7; 95(14):8268-73   
[0061]21. Hassen A, Belguith K, Jedidi N, Chemf A, Chemf M,
Boudabous A., Microbial characterization during composting of
municipal solid waste.   
[0062]22. Jager E, Ruden H, Zeschmar-Lahl B., [Composting
facilities. 1. Microbiological quality of compost with special
regard to disposable diapers], Zentralbl Hyg Umweltmed. 1994
October; 196(3):245-57. German.   
[0063]23. Jalees S., S. K. Sharma, S. J. Rahman and T Verghese,
1993. Evaluation of insecticidal properties of an indigenous
plant, Cannabis sativa L., against mosquito larvae under
laboratory conditions. J. Entomol. Res. 17:117-120.1993.   
[0064]24. Jenkins, Phil, "Field of Opportunity" Canadian
Geographic, Mar. 19, 1999   
[0065]25. Kane V V, Razdan R K. Constituents of hashish. A novel
reaction of olivetol with citral in the presence of pyridine.
Total synthesis of dl-cannabicyclol and dl-cannabichromene. J Am
Chem Soc. 1968 Nov. 6; 90(23):6551-3.   
[0066]26. Kashyap N. P., R. M. Bhagat, D. C. Sharma and S. M.
Suri, 1992. Efficacy of some useful plant leaves for the control
of potato tuber moth, Phthorimaea operculella Zell. in stores.
J. Entomological Research 16:223-227. 1992.   
[0067]27. Khan S M, Bennett J P Jr., Development of
mitochondrial gene replacement therapy., J Bioenerg Biomembr.
2004 August; 36(4):387-93. Review.   
[0068]28. Kir'yanova E. S. and E. L. Krall. Plant-Parasitic
Nematodes and their Control, Vol. II. Academy of Sciences of the
USSR, Nauka Publishers, Leningrad. 1971.   
[0069]29. Klingeren B. van and M. T. Ham. Antibacterial activity
of delta-9-tetrahydrocannabinol and cannabidiol. Antonie van
Leeuwenhoek 42:9-12., 1976.   
[0070]30. Kok C. J., G. C. M. Coenen, and A. de Heij, 1994. The
effect of fibre hemp (Cannabis sativa L.) on selected soil-borne
pathogens. J. International Hemp Association 1(1):6-9. [0071]31.
Kurilov V. I. and N. S. Kakhta. [More about hemp and the
Colorado beetle.] Zashchita Rastenii 1977 (7):63. 1977.  
 [0072]32. Lenehan N A, DeRouchey J M, Marston T T, Marchin
G L. Concentrations of fecal bacteria and nutrients in soil
surrounding round-bale feeding sites., J Anim Sci. 2005 July;
83(7):1673-9.   
[0073]33. Llewellyn G C, O'Rear C E., Examination of fungal
growth and aflatoxin production on marihuana., Mycopathologia.
1977 Dec. 16; 62(2):109-12.   
[0074]34. Loewe S., 1946. Studies on the pharmacology and acute
toxicity of compounds with marihuana activity. J. Pharmacology
and Expermental Therapeutics 88:154-164.   
[0075]35. Mackiewicz S., 1962. The effect of hemp on the density
of the potato beetle and the bean aphid. Biul. Ochrona Roslin
Inst. 16:101-131.   
[0076]36. Mateeva A. Use of unfreindly plants against root knot
nematodes. Acta Horticulturae 382 (February):178-182. 1995.  
 [0077]37. McPartland J. M., Fungal pathogens of Cannabis
sativa in central Illinois. Phytopathology 73:797., 1983,   
[0078]38. McPartland J. M. Pathogenicity of Phomopsis ganjae on
Cannabis sativa and the fungistatic effect of cannabinoids
produced by the host. Mycopathologia 87:149-153., 1984 [0079]39.
McPartland, John M., Cannabis as repellent and pesticide,
Journal of the International Hemp Association 4(2): 87-92, 1997
  
[0080]40. Mechoulam R, Hanu L. The cannabinoids: an overview.
Therapeutic implications in vomiting and nausea after cancer
chemotherapy, in appetite promotion, in multiple sclerosis and
in neuroprotection. Pain Res Manag 2001 Summer; 6(2): 67-73.   
[0081]41. Mechoulam R, Spatz M, Shohami E. Endocannabinoids and
neuroprotection. Sci STKE April 23; (129):RE5. 2002.   
[0082]42. Metzger F. W. and D. H. Grant. Repellency to the
Japanese beetle of extracts made from plants immune to attack.
USDA Technical Bulletin no. 299. 21 pp., 1932.   
[0083]43. Misra S. B. and S. N. Dixit. Antifungal activity of
leaf extracts of some higher plants. Acta Botanica Indica
7:147-150, 1979.   
[0084]44. Mojumder V, S. D. Mishra, M. M. Haque and B. K.
Goswami. Nematicidal efficacy of some wild plants against pigeon
pea cyst nematode, Heterodera cajani. Int. Nematol. Network
Newsletter 6(2):21-24, 1989.   
[0085]45. Nok A. J., S. Ibrahim, S. Arowosafe, et al. The
trypanocidal effect of Cannabis sativa constituents in
experimental animal try-panosomiasis. Veterinary and Human
Toxicology 36:522-524, 1994. [0086]46. S. Morimoto, F. Taura, Y.
Shoyama, Biosynthesis of cannabinoids in Cannabis sativa L,
Curr. Top. Phytochem. 2 (1999) 103-113.   
[0087]47. Oku, T. and Katsura, Y "Sequence analysis encoding
alpha-helix-turn-alpha-helix motif of HrpX in plant pathogenic
Xanthomonas", Unpublished   
[0088]48. Pandey K. N. Antifungal activity of some medicinal
plants on stored seeds of Eleusine coracana. J. Indian
Phytopathology 35:499-501.1982. [0089]49. Pandey J. and S. S.
Mishra. Effects of Cannabis sativa L. on yield of rabi maize
(Zea mays L.). in Abstracts of papers, Annual conference of
Indian Society of Weed Science. Bihar, India. 1982.   
[0090]50. Prakash A., I. C. Pasalu and K. C. Mathur, 1982.
Evaluation of plant products as paddy grain protectants in
storage. International J. Entomology 1:75-77.   
[0091]51. Prakash A., J. Rao and I. C. Pasalu, 1987. Studies on
stored grain pests of rice and methods of minimising losses
caused by them. Final Project Report (RPF-III) Ent-6/CRRI/ICAR
(India). 33 pp.  
 [0092]52. Quaghebeur K, Coosemans J, Toppet S, Compemolle
E, Cannabiorci- and 8-chlorocannabiorcichromenic acid as fungal
antagonists from Cylindrocarpon olidum., Phytochemistry. 1994
September; 37(1):159-61.   
[0093]53. Radosevic A., M. Kupinic and L. Grlic. Antibiotic
activity of various types of Cannabis resin. Nature
195:1007-1009. 1962.  
 [0094]54. Raharjo, Tri J; Chang, Wen-Te; Verberne,
Marianne C; Peltenburg-Looman, Anja M G; Linthorst, Huub J M;
Verpoorte, Robert, Cloning and over-expression of a cDNA
encoding a polyketide synthase from Cannabis sativa, Plant
Physiology and Biochemistry--Paris, 42 (4), 291-298, 2004.   
[0095]55. Riley C. V. and L. O. Howard. Hemp as a protection
against weevils. Insect Life (USDA) 4: 223 1892.   
[0096]56. Rothschild M., M. R. Rowan and J. W. Fairbairn.
Storage of cannabinoids by Arctia caja and Zonocerus elegans fed
on chemically distinct strains of Cannabis sativa. Nature
266:650-651. 1977.   
[0097]57. Schmitz J A, Olson L D., Duration of viability and the
growth and expiration rates of group E streptococci in soil.,
Appl Microbiol. 1973 February; 25(2):180-3.   
[0098]58. Shoyama, Y., Hirano, H., and Nishioka, I. (1978) J.
Labelled Ccmpd. Radiopharm. 14, 835-842 [0099]59. Sirikantaramas
S, Morimoto S, Shoyama Y, Ishikawa Y, Wada Y, Shoyama Y, Taura
F., The gene controlling marijuana psychoactivity: molecular
cloning and heterologous expression of
Delta1-tetrahydrocannabinolic acid synthase from Cannabis sativa
L., J Biol Chem. 2004 Sep. 17; 279(38):39767-74. Epub 2004 June
9.   
[0100]60. Stratii Y. I. Hemp and the Colorado beetle. Zashchita
Rastenii 5:61., 1976.   
[0101]61. Taura, F., Morimoto, S., Shoyama, Y., and Mechoulam,
R. (1995) J. Am. Chem Soc. 117, 9766-9767   
[0102]62. Taura, F., Morimoto, S., and Shoyama, Y. (1996) J.
Biol. Chem. 271, 17411-17416   
[0103]63. F. Taura, S. Morimoto and Y. Shoyama, Biosynthesis of
marihuana compounds--Purification and characterization of
biosynthetic enzymes. Current Topic in Plant Biology, Vol. 2 p
63-73 (2000), 2000.   
[0104]64. Turner C E, Elsohly M A., Biological activity of
cannabichromene, its homologs and isomers., J Clin Pharmacol.
1981 August-September; 21(8-9 Suppl):283S-291S.   
[0105]65. Van der Stelt M, Veldhuis W B, Bar P R, Veldink G A,
Vliegentharet J F, Nicolay K. Neuroprotection by
Delta9-tetrahydrocannabinol, the main active compound in
marijuana, against ouabain-induced in vivo excitotoxicity. J
Neurosci 2001 Sep. 1; 21(17): 6475-   
[0106]66. Van Haute, E., Joos, H., Maes, M., Warren, G., Van
Montagu, M., and Schell, J. EMBO J. 2, 411-417, 1983. [0107]67.

Van Klingeren B, Ten Ham M. Antibacterial activity of
delta9-tetrahydrocannabinol and cannabidiol., Antonie Van
Leeuwenhoek; 42(1-2):9-12 1976. [0108]68. Veliky I. A. and R. K.
Latte. Antimicrobial activity of cultured plant cells and
tissues. Lloydia 37:611-620., 1974. [0109]69.   
Vijai P., I. Jalali and R. D. Parashar. Suppression of bacterial
soft rot of potato by common weed extracts. J. Indian Potato
Association 20:206-209, 1993. [  
0110]70. White, F. F., and Nester, E. W. J. Bacteriol. 141,
1134-1141, 1980.   
[0111]71. T. Yamaguchi, F. Kurosaki, D.-Y. Suh, U. Sankawa, M.
Nishioka, T. Akiyama, M. Shibuya, Y. Ebizuka, Cross-reaction of
chalcone synthase and stilbene synthase overexpressed in
Escherichia coli, FEBS Lett. 460 457-461, 1999.   
[0112]72. Zelepukha S. I., 1960. The third conference on the
problem of phytoncides. J. Mikrobiol, Kiev 22(1):68-71.A

---