HOW TO CHOOSE THE APPROPRIATE RECOMBINING MICROORGANISM

FOR YOUR BIOPRODUCTION

One of the questions that is mandatory and sometimes becomes a real headache is: "How do
I choose the best recombinant microorganism for what I want to produce in my industry?" ...
and the answer to this question, as you know , is of fundamental importance for you and your
company.

You must choose very carefully the recombinant microorganism that you are going to use,
since this will largely determine the productive success of your manufacture. Additionally,
you have to take into account that genetically transformed strains are very expensive and,
from this point, you will start your monetary investment, which you must take very good care
to sustain the profitability of your business.

With this in mind, let's get into the matter ...

INDEX.
• What is a recombinant microorganism.
• A little history about Recombinant DNA.
• Recombinant microorganisms and their role in the contemporary industry.
• Criteria for selecting a recombinant microorganism.
• Conclusions.
• Bibliography.
WHAT IS A RECOMBINANT MICROORGANISM.
Basically, a recombinant microorganism is one that has been genetically modified to
give rise to new microbial products. This genetic transformation occurs through the
application of recombinant DNA techniques, which consist, in broad strokes, of
inserting pieces of foreign DNA into the DNA of a microorganism. In this way, the
replication and expression machinery of that microorganism is used to give rise to the
product of interest.

A LITTLE HISTORY ABOUT RECOMBINANT DNA.

It all started in 1972, when a student from the Department of Biochemistry at the Stanford
University School of Medicine first proposed the idea of recombinant DNA. The name of this
young man, a graduate of Professor Dale Kaiser, is Peter Lobban.

In the years from 1972 to 1974, the technique was carried out by Lobban and Kaiser, Jackson,
Symons and Berg and Stanley Norman Cohen, Chang, Herbert Boyer and Helling. And, in
parallel, between 1972 and 1973, the first scientific publications on the production and
intracellular replication of recombinant DNA appeared. These findings fundamentally
described the techniques for isolating and amplifying genes or segments of DNA and
insertion techniques for the creation of a transgenic bacterium.En 1978, Werner Arber, Daniel
Nathans and Hamilton Smith received the Nobel Prize in Medicine for new contributions made
in this area, which consisted of the discovery, isolation and application of restriction
endonucleases.

In 1980, the patent requested in 1974 on the process for the production of biologically
functional molecular chimeras, which could not exist in nature, was finally granted to the
University of Standford, in the people of Cohen and Boyer.

And finally in 1982, the Genetech company, authorized by Eli Lilly and Company, developed the
first legal biopharmaceutical, human insulin, using recombinant DNA technology.

From then on, a lot of water has run under the bridge.

RECOMBINANT MICROORGANISMS AND THEIR ROLE IN THE

CONTEMPORARY INDUSTRY.
Most of the work in industrial microbiology is currently being carried out through the use of
microorganisms modified by genetic mutations, which I will call throughout the post
"genetically transformed microorganisms" or, to abbreviate even more its name,
"recombinant microorganisms".

Modern biotechnology, through recombinant DNA techniques, has given rise to

microorganisms with the desired genetic characteristics for the intended purposes. And this
has been a breakthrough in the industry, replacing slower and low performance
biotechnology techniques, with faster, more direct and more efficient techniques for
obtaining the products of interest.

Genetically transformed microorganisms or recombinant microorganisms are currently

used in industrial microbiology to create a wide variety of products. Among them: enzymes
for various uses and applications, vitamins, antibiotics, vaccines and recombinant insulin,
acidifying agents, biosurfactants, emulsifiers and biofuels in general. As you can see, the
food, pharmaceutical, agricultural and other industries, such as the textile, chemical, etc., are
directly involved in this trend.

The products of industrial microbiology, called "recombinant products", today have a great
impact on all aspects of our lives: antibiotics and other microbial products continue to
contribute to the welfare of animals and humans; bacteria; fungi and viruses are being used
as biopesticides, reducing dependence on chemical pesticides; We can also include anti-
tumor compounds, hybrid antibiotics and biosensors development. Anyway. And this is a
trend that will not stop.

If you want to “get on the train” of bioproduction for the first time, or if you are opening a new
recombinant production line, you will be interested in knowing the criteria to select the most
appropriate strain for your interests.

CRITERIA TO CHOOSE THE MOST APPROPRIATE RECOMBINING

MICROORGANISM.
I told you that it is super important and of fundamental importance to have a clear criterion
when selecting the recombinant microorganism that meets, but exactly, at least as closely as
possible, the conditions necessary for your production interests. Next, I will mention some
key evaluation parameters:

• Biological system.
You must ensure that you acquire a biological system that is well known, that is, that is well
characterized and, additionally, that is easily cultivable. Among the biological systems that
can be used are bacteria, yeasts and recombinant mammalian cells.

• Speed or production time.

The speed of the processes is a parameter of great interest at the level of industrial
production. Therefore, you must take into account that, depending on the biological system
chosen and due to its intrinsic biological characteristics, its growth rate and production of the
recombinant product will vary. You should know that bacteria are usually faster than yeasts
and both faster than mammalian cells.

• Yield or large-scale production capacity.

Due to the degree of stability or robustness presented during its growth and its speed of
growth and production, we can discriminate whether we are facing the best biological system
to produce on a large scale. It is known that bacteria and yeasts have a high capacity for large-
scale production, but mammalian cells do not.
Note: Here it is also important to determine the size of the productive batches that one aspires
to obtain and whether that recombinant product must necessarily be produced on a large
scale. It is also necessary to determine if the recombinant product that interests us can be
produced by any biological system or not.

• Quality of the final product.

Understanding as a final product the recombinant product and attributing "quality" to the fact
that it is as similar as possible to the original product in terms of structure and function, it is
important to evaluate the importance of this criterion in the selection of the biological system
to be used. In this sense, bacteria and yeasts are classified as intermediate quality compared
to the high quality of mammalian cells.

• Contamination risks
This criterion is very important to study in advance, since it determines controls that will have
to be introduced to the production process with the costs that this entails and the risks
involved in losing entire batches due to contamination. Bacteria usually have risks of
contamination by endotoxins and mammalian cells by viruses and prions, but yeasts have a
low risk of contamination, which could in any case be caused by bacterial contamination
during cultivation.

• Genetic stability
Genetic stability is nothing more than the tendency to keep the genotype stable over time. It is
also important that our biological system brings us that security. It would be worthwhile to
acquire a very genetically transformed biological system, but that presents ease of losing its
post transformation genotypic characteristics.

• Cost
It is no accident that you put this parameter last. I do it this way, because at the time of
acquiring a biological system for our productive process, costs should not be spared. If all the
previous points (and some more) are evaluated, a certain recombinant strain is favorable, it
is best to pay it at any cost. However, if this is not possible, then a second best option should
be available… but always giving priority to the aforementioned criteria.

Conclusions
Acquiring a recombinant strain in order to produce on an industrial level is a very important
issue. If the choice of the strain is not the best, if the strain is not ideal for the purposes and
purposes expected of it, it can eventually mean a problem that will creep over time and that
may lead to a decrease of profitability and even economic losses. That is why you must
establish your bioproduction objectives very well and analyze each one of the parameters
mentioned in this post in order to create a criterion that guides you and allows you to acquire
the ideal recombinant microorganism or at least as adjusted to your requirements possible.

I hope you enjoyed this post, that you found interesting, useful and, above all, that it serves as
an orientation and a starting point to improve your business.

Regards,
Liliana Berlioz.
President and Executive Director.
Biotech Consulting, LLC.
www.lilianaberlioz.com
Bibliografía.
 Prescott, L. Harley, J. Klein, D. (2002). Microbiología. España, Madrid. McGraw Hill.
 Argenbio. “Las proteínas recombinantes en la industria”. Argenbio,
http://www.argenbio.org/index.php?action=novedades&note=245. Accedida el
21/08/2019.
 María Iranz. “Bioproductos”. María Iranz. Blog de una biotecnóloga.
http://www.mariairanzobiotec.com/bio-productos/. Accedida el 21/08/2019.
 José Antonio Villaverde. “ADN recombinante”. El enciclopedista. 28/10/2012.
http://www.elenciclopedista.com.ar/adn-recombinante/.
 Wikipedia. “Tecnología de ADN recombinante”. Wikipedia. 27/07/2019.
https://es.wikipedia.org/wiki/Tecnolog%C3%ADa_de_ADN_recombinante.
 Principios de Biotecnología. “Principios de Biotecnología”. Principios de Biotecnología.
26/04/2013. http://caiagronomia.blogspot.com/.
 ADN recombinante. “Historia ADN recombinante”. ADN recombinante. 12/10/2016.
http://dnarecombinanteud.blogspot.com/2016/10/historia-adn-recombinante.html.
 ArgenBio. “Las proteínas recombinantes en la industria”. ArgenBio.
http://www.argenbio.org/index.php?action=novedades&note=245. Accedida el
22/08/2019.