THE NATIONAL TEACHERS COLLEGE

629 J. Nepomuceno St. Quiapo, Manila

School of Advanced Studies

Subject: Special Topics in Biological Sciences

Prepared by: Romar Vincent D. Cabutaje
Professor: Dr. Amparo Sulam

SYNTHESIS:
ROLES OF FUNGI IN THE ENVIRONMENT
Fungi play a crucial role in the balance of ecosystems. They colonize most habitats on
earth, preferring dark, moist conditions. They can thrive in seemingly-hostile
environments, such as the tundra. However, most members of the Kingdom Fungi grow
on the forest floor where the dark and damp environment is rich in decaying debris
from plants and animals. In these environments, fungi play a major role as decomposers
and recyclers, making it possible for members of the other kingdoms to be supplied with
nutrients and to live.
The food web would be incomplete without organisms that decompose organic matter.
Some elements, such as nitrogen and phosphorus, are required in large quantities by
biological systems; yet, they are not abundant in the environment. The action of fungi
releases these elements from decaying matter, making them available to other living
organisms. Trace elements present in low amounts in many habitats are essential for
growth, but would remain tied up in rotting organic matter if fungi and bacteria did not
return them to the environment via their metabolic activity.
The ability of fungi to degrade many large and insoluble molecules is due to their mode
of nutrition. As seen earlier, digestion precedes ingestion. Fungi produce a variety of
exoenzymes to digest nutrients. These enzymes are either released into the substrate or
remain bound to the outside of the fungal cell wall. Large molecules are broken down
into small molecules, which are transported into the cell by a system of protein carriers
embedded in the cell membrane. Because the movement of small molecules and
enzymes is dependent on the presence of water, active growth depends on a
relatively-high percentage of moisture in the environment.
As saprobes, fungi help maintain a sustainable ecosystem for the animals and plants
that share the same habitat. In addition to replenishing the environment with nutrients,
fungi interact directly with other organisms in beneficial, but sometimes damaging,
ways.
ROLES OF FUNGI IN THE INDUSTRY
Many fungi are useful to humans and have been exploited both industrially and
commercially. Societies have utilized fungi for centuries in a wide variety of ways by
capitalizing on the metabolism and metabolites (chemicals made from metabolism)
produced. The oldest and best known example is the use of yeasts performing
fermentation in brewing, wine making and bread making. Yeasts and other fungi play a
critical role in drug production, food processing, bio-control agents, enzyme
biotechnology, as well as research and development.
The use of yeast (e.g. Saccharomyces cerevisae) to make alcohol and carbon dioxide
uses the fermentation process to break down sugars. Up to 50% of the sugar can be
converted to alcohol, but rarely surpasses 15% because the fungi are sensitive to high
concentrations of alcohol. In the beer making industry, cereal grains are fermented to
make the final product. Wine is composed of fermented grapes while hard cider is
essentially fermented apples. Sake is produced by rice fermentation, using Aspergillus
oryzae and then an additional fermentation step utilizing bacteria and yeasts. With
bread making, fermentation utilizes sugar to produce carbon dioxide and alcohol. The
carbon dioxide produces the bubbles and causes bread to rise, while the alcohol
produced evaporates off while baking. A variety of foods and drinks are fermented
before consumption. Some of the largest industries in the west are based around
production of alcohol and bread. In the east, the production of soy sauce and other
fermented soybean products are likely among the largest industries. The peoples of Asia
have developed a wide variety of interesting fermented foods, sauces and drinks, using
fungi. Other examples and the applicable fungi include koji (Aspergillus); miso, soy bean
paste (Aspergillus); sufu, Chinese cheese (Rhizopus), nyufu or fuyu, bean cake or bean
cheese (Rhizopus); shoyu or soy sauce (Aspergillus, Saccharomyces) and tempeh
(Rhizopus).
Another way in which fungi are used industrially in the food industry is in cheese
production. Various cheeses are inoculated with Penicillium roquefortii to impart a
strong and pungent flavor in the resultant cheeses. Examples are Roquefort, Gorgonzola,
Stilton Blue and Danish Blue. The white crust on the outside of the cheeses known as
Brie and Camembert is the mycelium of Penicillium camembertii. These strong flavors
are a result of the fungus producing methyl ketones.
Recombinant Technology and its Application: GENE THERAPY

Recombinant DNA technology has wide spectrum of applications in treating diseases

and improving health conditions. In the past century, the recombinant DNA technology
was just an imagination that desirable characteristics can be improved in the living
bodies by controlling the expressions of target genes. However, in recent era, this field
has demonstrated unique impacts in bringing advancement in human life.

By virtue of this technology, crucial proteins required for health problems and dietary
purposes can be produced safely, affordably, and sufficiently. This technology has
multidisciplinary applications and potential to deal with important aspects of life, for
instance, improving health, enhancing food resources, and resistance to divergent
adverse environmental effects. Particularly in agriculture, the genetically modified
plants have augmented resistance to harmful agents, enhanced product yield, and
shown increased adaptability for better survival.

Moreover, recombinant pharmaceuticals are now being used confidently and rapidly
attaining commercial approvals. Techniques of recombinant DNA technology, gene
therapy, and genetic modifications are also widely used for the purpose of
bio-remediation and treating serious diseases. Due to tremendous advancement and
broad range of application in the field of recombinant DNA technology, this review
article mainly focuses on its importance and the possible applications in daily life.

Gene therapy typically involves the insertion of a functioning gene into cells to correct a
cellular dysfunction or to provide a new cellular function (Culver, 1994). Gene therapy
can be used to correct or replace the defective genes responsible. Gene therapy has
been especially successful in the treatment of combined immunodeficiency syndromes,
showing lasting and remarkable therapeutic benefit.

GENE THERAPHY
A. What is gene therapy? Why is it used?
 an advanced technique with therapeutic potential in health services
 involves the insertion of a functioning gene into cells to correct a cellular
dysfunction or to provide a new cellular function (Culver, 1994)
 an advanced technique with therapeutic potential in health services. The first
successful report in field of gene therapy to treat a genetic disease provided a more
secure direction toward curing the deadliest genetic diseases
 GT is used to correct a deficient phenotype so that sufficient amounts of a normal
gene product are synthesized to improve a genetic disorder.
B. How is Gene Therapy Carried Out?
 Modification of somatic cells by transferring desired gene sequences into the
genome.
 Somatic cells necessary to ensure that inserted genes are not carried over to the
next generation.
C. Different Delivery Systems are Available
 In vivo versus ex vivo
 In vivo = delivery of genes takes place in the body
 Ex vivo = delivery takes place out of the body, and then cells are placed back
into the body
 In vivo techniques usually utilize viral vectors
 Virus = carrier of desired gene
 Virus is usually “crippled” to disable its ability to cause disease
 Viral methods have proved to be the most efficient to date
 Many viral vectors can stable integrate the desired gene into the target cell’s
genome
 Problem: Replication defective viruses adversely affect the virus’ normal
ability to spread genes in the body
 Reliant on diffusion and spread
 Hampered by small intercellular spaces for transport
 Restricted by viral-binding ligands on cell surface therefore cannot advance
far.
 Ex vivo manipulation techniques
 Electroporation
 Liposomes
  Calcium phosphate
 Gold bullets (fired within helium pressurized gun)
 Retrotransposons (jumping genes – early days)
 Human artificial chromosomes
D. Limitations of Gene Therapy
 Gene delivery
 Limited tropism of viral vectors
 Dependence on cell cycle by some viral vectors (i.e. mitosis required)
 Duration of gene activity
 Non-integrating delivery will be transient (transient expression)
 Integrated delivery will be stable
 Gene control/regulation
 Most viral vectors are unable to accommodate full length human genes
containing all of their original regulatory sequences
 Human cDNA often used much regulatory information is lost (e.g. enhancers
inside introns)
 Often promoters are substituted therefore gene expression pattern may be
very different
 Random integration can adversely affect expression (insertion near highly
methylated heterogeneous DNA may silence gene expression)
 Expense
 Costly because of cell culturing needs involved in ex vivo techniques
 Virus cultures for in vivo delivery
 Usually the number of patients enrolled in any given trial is <20
 More than 5000 patients have been treated in last ~12 years worldwide
E. Case of JESSE GELSINGER
 first person publicly identified as having died in a clinical trial for Gene Theraphy.
 suffered from Ornithine Transcarbamylase Deficiency
 symptoms: inability to metabolize amonia.
 had a milder form of disease in which the OTD is mutated in only part of the
patient's cell, a condition is called SOMATIC MOSAICISM
 he joined a clinical trial by the University of Pennsylvania
 On Sept. 13, 1999, he was injected with an Adenoviral Vector
 he died 4 days later at the age of 18, apparently having suffered a massive immune
response triggered by the use of Viral Vector to transport gene into his cell leading
to multiple organ failure and brain death.
 According to FDA investigation, they concluded that the scientist involved in the
trial, including the co-investigator DR. JAMES M. WILSON (Director of the Institute
for Human Gene Therapy) broke several rules of conduct such as:
 Inclusion of Gelsinger as a substitute for another volunteer who dropped out,
despite Gelsinger's having high amonia levels that should have led to his
exclusion from the trial.
 Failure by the University to report that two patients had experienced serious
side effects from the gene theraphy.
 Failure to disclose, in the informed-consent documentation, the deaths of
monkeys given a similar treatment.