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INDUSTRIAL SECRETS OF L-EPHEDRINE PRODUCTION

Chapter 11
Dedication: This publication is dedicated to the head of the DEA Chuck
Rosenberg, head of the Sinaloa Cartel Joaquin Guzman (El Chapo), his attorneys
Juan Pablo Badillo and Jose Refugio Rodriguez, as well as the numerous victims
of a futile and fragile drug war. None offered anything to prevent publication of
these books.
11. Optimizing Yeast Based on Darwins Theory of Evolution

Some publications available on the Internet state that yeast was optimized for L-PAC
production by using ultraviolet rays to induce mutations in the yeast. Although this may
have some merit, this procedure for the most part would serve to produce UV tolerant
yeast, which would be useless in the benzaldehyde biotransformation to L-PAC.

Microorganisms evolve to resist substances that are toxic to their survival, such as
bacteria has evolved to resist the use of antibiotics. Microorganisms are known to evolve
relatively quickly, especially when they are not completely wiped out and the strongest
survive to breed the next generation. This is an occasional occurrence when patients do
not finish an antibiotic cycle, leaving only the strongest bacteria cells. These
strong/resistant cells now breed a new generation, which has lead to super bacteria
and flesh eating diseases that do not respond to conventional antibiotic treatment.

The correct procedure to optimize yeast is to build


or purchase a laboratory sized stirred tank fermenter
(bioreactor), with a self healing inoculation port.
The port may simply be a self sealing, synthetic
rubber port in which needles can be inserted to
inoculate the fermenter or obtain samples from the
bioreactor/fermenter.

A sterile yeast sample should be taken using a sterile


needle near the end of a biotransformation process
of benzaldehyde in a laboratory size fermenter, as

the strongest yeast cells will be the


last that survive. Using an alcohol
swab to sterilize the needle and
inoculation port while extracting
or injecting a sample is highly
recommended.

The needle for

obtaining a sample should be half


filled

with

sterile

6%

dextrose/water, to immediately
lower the toxic levels of L-PAC,
benzaldehyde,

aldehyde,

and

benzyl alcohol that are now killing the yeast, while providing sustenance for the yeast
to grow. This sample should be taken when evolving carbon dioxide has slowed to a
fraction of the gas production peak. The sample should be inoculated onto a sterile
nutrient agar medium in multiple Petri dishes located in a clean room or clean box,
so that these remaining yeast cells may reproduce on the Petri dishes. It is possible to
go too far and have killed/sterilized all the yeast cells before inoculation, so timing with
obtaining the sample is a matter of trial and error. I would recommend for most strains,
a 50% reduction in carbon dioxide output would be a good time to acquire a sample, but
if one is able to take it further to 25% or less and still have remaining viable yeast cells
that breed on the agar, then I would do so as to more rapidly force the evolution.

This is an Example of a Petri Dish Inoculated


By Moving the Needle From Side-to-Side

After several days of incubation at 25C+,


the fastest growing mycelium among the
Petri dishes should be used to inoculate
another batch for L-PAC production within
the laboratory size bioreactor. As the
contents of the Petri dish wont easily inject
through a needle, the use of a sterilized
tissue

homogenizer

will

assist

with

breaking up the yeast and agar into an


injectable solution. All of this should be
performed in a clean room or clean box
to prevent contamination.
The above procedure of small L-PAC
production runs and growing the yeast that
is the last to die should be repeated many
times. Sometimes but not always, different strains of the same type of yeast should be
introduced to encourage sexual reproduction of the yeast and to increase the genetic
potential. Slowly through many batches, the yeast will begin to show tolerances through
a simple but rapid evolutionary process, where the yeast will eventually reach a genetic
peak that cannot be surpassed.

The genetic peak that is reached, where no more gains can be made, is your optimized
yeast. I recommend using high alcohol tolerant strains of Saccharomyces Cerevisiae as
base strains, which are available from any home brew supply store. Although some
literature suggests the use of Candida Utilis has resulted in very high yields, I believe
this may be a publication designed to once again mislead the public. We have found
Candida Utilis to have little to no tolerance to alcohol production, where other
documents recommend Candida Flareri, which can be purchased from a variety of
yeast banks on the Internet. At the end of the day, experiment with any yeast you
want, but S. Cerevisiae bread for alcohol production will work for certain.

After reaching this stage of a genetic peak, UV-C light exposure can then be attempted
to see if any mutations can be induced that would further increase the optimization of
the strain, but I recommend this only after the genetic peak of evolution has been
reached.
Other means of developing an optimized mutant yeast is described in patent:

Process for making phenyl acetyl carbinol (pac), microorganisms for use in the
process, and a method for preparing the microorganisms
EP 0445129 A1
PCT number

PCT/US1989/004423

I have often wondered how easy it would be to simply purchase an optimized yeast
sample from an employee of an Indian or Chinese ephedrine producer. India and China
have low wages compared to western dollars, therefore many employees may be more
than willing to provide a sample in exchange for perhaps a few thousand dollars. This
may save a producer a large amount of work in having to perform their own yeast
optimization.

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