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Vitamin K2

Chrestina Abounaoum, Ester Fernandez, Rebecca Graff, Megan Hupp, Britt Robinson,
Alexandra Stronge

Introduction
Vitamin K may not be very well known, but it is a nutrient that is able to make a
big difference for a healthy heart and bones (Jacob 2013). There are three known types
of vitamin K: vitamin K1(phylloquinone), vitamin K2(menaquinone) and vitamin
K3(menadione) (Mercola 2004). Vitamin K1 can be found in green vegetables and is
known to help maintain healthy blood clotting by going straight to the liver (Ramirez
2014). Vitamin K3, menadione, is a synthetic form, which is not recommended to take
because it has been known to cause toxicity in infants who have been treated with it
(Mercola 2014). Vitamin K2, menaquinone, is primarily found in the bacteria that line the
gastrointestinal tract but goes straight to the walls of blood vessels, bones, and other
tissues than your liver (Mercola 2014). Throughout this paper some benefits of vitamin
K2, the chemistry and biological functions of the molecule, the sources and the
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deficiencies caused by the lack of the vitamin and some research that has been done
will be discussed.
Vitamin K2 is a fat-soluble vitamin and therefore, for proper absorption, some fat
must be eaten along with it (Leech 2015). Vitamin K2 is a vitamin that is able to provide
many benefits to those who consume it. One of the most prominent being that it helps to
prevent the hardening of arteries, which are common side effects in coronary artery
disease and heart failure (Ramirez 2014). Research has shown that the vitamin may
even help to keep calcium out of places where it can cause damage, such as body
tissues and artery linings (Mercola 2004). Menaquinone is also very important in
improving bone density, since it is able to pull calcium and other important minerals into
the bone matrix (Mercola 2004). It has been found that Vitamin K2 slowed the growth in
cancer cells in patients with lung cancer and treating leukemia (Aoshima, et al., 2003). A
German research group in 2008 discovered that vitamin K2 is able to provide protection
against prostate cancer (Linseisen, et al., 2008). It has also been found to be helpful
against non-Hodgkin lymphoma, oral cancers, nasopharynx, stomach and colon cancer
(Mercola 2004). Researchers have also seen evidence that vitamin K2 can help with the
development of Alzheimers disease and with type 2 diabetes (Allison, et al., 2001). The
research that has been done, has shown the many benefits that the consumption of
vitamin K2 can offer.

Chemistry & Discovery of Vitamin K2

Vitamin K2 is a general term for all menaquinones. Menaquinones, or MKs for
short, are known as isoprenologues. Isoprenologues have a 2-methyl-1,4-
napthoquinone with differing polyisoprenoid side-chains (Booth, et. al., 2013). The
napthoquinone is composed of two fused benzene rings with oxidized carbons at the 1
and 4 positions. Napthoquinone is a derivative of naphthalene. The polyisoprenoid side-
chains can range anywhere from 5-13 units. Isoprenoids are derivatives of isoprenes or
terpenes. The names of the menaquinones differ depending on the length of the
isoprenoid side-chains. For example, there is Menaquinone-4 (or MK-4) which has four
isoprenoid side chains and Menaquinone-5 (or MK-5) which has five isoprenoid side
chains (Buelens, et. al., 2013).
The most abundant forms of menaquinone in the human diet are MK-4, MK-7,
MK-8, MK-9, and MK-10. MK 4 is considered a short-chain menaquinone whereas MK-7
through MK-10 are considered long-chain menaquinones (Buelens, et. al., 2013).
Mostly all of the menaquinones are synthesized by bacteria; the only exception to this is
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MK-4 which is made in the human body through the conversion of the synthesized
Vitamin K3 (Booth, et. al., 2013).
Vitamin K and menaquinones were first discovered in 1931 at the Ontario
Agriculture College. There was an experiment done by some scientists that involved
chickens and chicken feed. These experimental chickens were commonly fed fish meal
and the scientists originally extracted the fish meal with ether, but they noticed the baby
chicks were hemorrhaging. However, when the fish meal was extracted with water and
then allowed to ferment, the chicks did not hemorrhage. This was because the ether
actually removed all of the bacteria that could synthesize the menaquinones. It was later
discovered that Vitamin K2 was present within the fish meal and responsible for the
differences in hemorrhaging. For a while, putrefied fish
meal was the best way to get this mysterious compound (Bentley, et. al., 1982).
Many bacteria are capable of synthesizing menaquinones. Bacillus cereus,
E.Coli, and Staphylococcus aureus are just a few examples. This synthesizing property
was discovered over time by growing these different bacteria on agar plates and then
Vitamin K was found on the plates (Bentley, et. al., 1982). Bacteria synthesize
menaquinones through a complex chemical pathway. Within the bacteria there is a
certain compound called shikimate. Shikimate gets its interesting name from the
Japanese Star Anise flower (or shakimi flower) that it was first isolated from. The
shikimate pathway is normally used in bacteria to biosynthesize certain aromatic amino
acids that they need, like phenylalanine, tryptophan, and tyrosine. If youve taken
genetics, then you know that bacteria are prototrophs which means that they make
everything they need to survive. The shikimate pathway is an example of how they do
that with certain amino acids. The menaquinone-synthesizing pathway in bacteria starts
with shikimate and then is converted to chorismate. Through a series of steps and
different intermediates, including alpha-ketoglutarate (a byproduct of working with amino
acids) we end up with dimethylmenaquinone (or DMK) and then eventually
menaquinone (Bentley, et. al., 1982).

Brief Overview of Function of Vitamin K2

The biological function of vitamin K2 begins with bacteria, before it begins in
humans. As with people, bacteria also need to synthesize energy (although not on such
a large a scale), and they use similar pathways to make ATP: namely, bacteria use an
electron transport pathway that requires the use of redox reaction-available quinones.
Unlike animals, which use ubiquinone, bacteria synthesize menaquinones, which show
a similar function in bacterial electron transport (Das, et al, 1989).
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When introduced to the human system, menaquinones (the K vitamins) take on a

very different role. Rather than being used for energy production, these menaquinones
interact with vitamin K-dependent proteins, thus named due to their particular interaction
with vitamin K. Vitamin K is seen to be a cofactor in the enzyme gamma-
carboxyglutamate carboxylase, which converts glutamate residues (Glu) in some
proteins to gamma-carboxyglutamate (Gla) this specific interaction of gamma-
carboxyglutamate. There are seventeen proteins in this family known to date, two of
which are focused on here for their major physiological importance: osteocalcin (OC)
and matrix Gla protein (MGP)(Beulens, et al, 2013).
OC is a small protein existing in bone tissue, which makes up 0.5-1.0% of bone
proteins, and 20% of noncollagenous protein. OC exhibits vitamin K dependency with
calcium-binding Gla residues. When activated, this protein binds two moles of calcium
per 6,500 g of OC (Lian, et al, 1978). The purpose of this protein is to bind calcium and
deposit it in the bone matrix, thus fortifying and hardening the bone tissue.
MGP functions in a similar way to OC, in that it relies on post-translational
modification by vitamin K-dependent carboxylase to become active (Glu residues are
converted to Gla residues) (Schurgers, et al, 2007). MGP differs from OC in terms of
body location: MGP is expressed by vascular smooth muscle cells as well as bone,
dentine, and other soft tissue cells. Additionally, MGP functions in a similar way to OC,
by mobilizing calcium, however it is seen to inhibit calcification of soft tissue and blood
vessels, rather than promoting it (Beulens, et al, 2013).
Deficiency of Vitamin K2
Vitamin K2 deficiency can cause many issues such as coronary vascular
disease, heart attack, stroke, cancer, osteoporosis, and arterial calcification (Mercola
2012). Osteoporosis and arterial calcification are the two diseases that have been
studied the most in relation to vitamin K2. Vitamin K2 deficiency is defined as an
increase in the concentration of undercarboxylated osteocalcin (OC) which is a protein
produced by osteoblasts (Beulens, et al., 2013). Vitamin K2 is essential for the
carboxylation of OC and its accumulation in osteoblasts (Beulens, et al., 2013).
Osteoporosis occurs when the mass of the bones is decreased which causes
them to be very brittle. Osteoclasts, which degrade bone (known as resorption) are
doing their job faster than osteoblasts that deposit calcium to rebuild the bone (Beulens,
et al., 2013). Vitamin K2 has been found to induce osteoclast apoptosis, so cells that
degrade bone are diminished, allowing osteoblasts to work more efficiently. Vitamin K2
also sustains lumbar bone mineral density to prevent osteoporosis-related bone
fractures in elderly patients (Iwamota, Sato, Takeda, 2004). One experiment found that
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serum levels of vitamin K2 were lower in patients with osteoporosis and osteoporotic
fractures compared with controls (Aoki, et al., 2000). It was also found that vitamin D
may play a role in the effectiveness of vitamin K2 (Mercola, 2012). Usually doctors tell
their patients to take calcium supplements to help with osteoporosis, however as more
information about vitamin K2 becomes available, researchers are finding that taking
calcium alone may have no effect on osteoporosis, but a negative one on your arteries
(Chierici, et al., 2013).
If a person is deficient in vitamin K2, they should not take calcium supplements
as calcium will build up in the arteries causing calcification without vitamin K2 to take
calcium to the bones (Jacob, 2013). Calcification leads to problems like cardiovascular
disease (CVD). MGP is a protein that inhibits vascular calcification when it is
carboxylated by K2, reducing the risk of CVD (Jacob, 2013). Studies show that a lower
intake of K2 produces more severe aortic calcifications, where a higher intake produces
more mild calcifications (Beulens, et al., 2013).
It is difficult to differentiate between the effects of Vitamin K1 and K2 in the body,
so studies must use the method of direct intake of K2 in order to study its effects. For
this reason, there have been very few experiments to test the amount needed in our
diets (Mercola, 2012).Though it is possible for gut bacteria to produce vitamin K2, it is
not enough to prevent deficiency (Mercola, 2012). There are only estimated values of
intake right now range from 33 to 45 mcgs,
which have only been tested on patients with
osteoporosis (Beulens, et al., 2013).

Vitamin K2 Studies
Many studies have been conducted
using Vitamin K. These studies contain a
wide array of topics. The goals of these
topics are to understand the role of Vitamin
K2 in cardiovascular and bone health, find
treatments for cancer, and gain a better understanding for the way that menaquiones
function.
In the Netherlands a study was conducted to prove that Vitamin K2 can be used
to help reduce the risk of developing heart disease. The statistics of this study are done
based on the participants intake of Vitamin K2. Participants that were in the upper third
of intake of Vitamin K2 showed a 52% decrease in arterial calcification. The way that
Vitamin K was able to accomplish this drastic decrease was by working as a leader for
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calcium. It would lead the calcium in the body away from the arteries and help the
calcium get to the bones. Due to Vitamin K2s ability to keep calcium away from the
blood vessels the study that was conducted also showed a 41% decrease in the
development of heart disease and a 57% decrease in death from heart disease.
After helping the Calcium get out of the arteries Vitamin K2 continues helping to
lead calcium by getting the calcium to the bones. Vitamin D causes our body to create
K2 dependent proteins. These proteins are used to help lead calcium to the right areas
of the body. K2 comes in to activate these proteins and thus strengthening the
interaction between Calcium and Vitamin D. Prior to the understanding of Vitamin K2s
role in this process, studies had come out stating that increased calcium in a consumer
diet put a person at risk for stroke.
A study aiming to understanding Vitamin K2s effect on cancer identified two
mechanisms that helped with the treatment of cancer.The first is to induce cell death
with the help of cotylenin A. This experiment was proven effective in Human Leukemia
HL-60 Cells. In figure 1 we can see the data from this experiment. In A we can see that
cell proliferation was the greatest because it contained neither Cotlyn A or Vitamin K2. B
contained just Cotlyn A and C contained just Vitamin K2. When we put Cotlyn
A and Vitamin K2 together we see the least amount of cells being created.
The second method that Vitamin K2 uses to fight off cancer is through
autophagy. This is a system the body uses to keep cell proliferation in balance. When
Leukemia cells were treated with K2 autophagosomes formed. These helped to regulate
cell death when the cells were not in immediate danger of death. These two different
mechanisms apoptosis and autophagy work together to best fight against cancer.
Apoptosis in times of immediate danger and autophagy in times when long term
regulation can be focused on.
Other research that is being done has lead to the discovery of 5 genes
responsible for Menaquinone synthesis. These were discovered by working with mutant
e.coli strains on glucose media.Scientists then would look to see which phenotypes
were not expressed when any of these 5 genes were mutated. These experiments also
proved that menaquinones have a role in aerobic metabolism. This is done by the
menaquinone working as electron acceptors.

Sources of Vitamin K2
From supplements to food, there are many sources of Vitamin K2. As we
mentioned before the gut is able to synthesize Vitamin K2 from Vitamin K1 (Kresser,
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2008). While its true our gut microbiomes have the ability to synthesize Vitamin K2,
there are many studies that suggest the gut cannot produce sufficient amounts of K2 for
it to be beneficial to our bodies. Another source for Vitamin K2 resides within the food
we eat.
While Vitamin K1s food sources mainly reside in leafy green vegetables
(dandelion and mustard greens, swiss chard etc.) Vitamin K2s food sources are largely
obtained from meats (chicken liver, beef and salami), soft and hard cheeses, and eggs
(National Academies Press, 2002). However Vitamin K2 is not only found in dairy and
meat sources, but also in a vegan variety.
Natto is a traditional Japanese food, it is soybean fermented with Bacillus subtilis
var. natto. A single serving of Natto (100g) will provide you with 23 mcg of Vitamin K2
(which is 29% of the daily value of K2 in Japan). Its rich in the MK-7 variation of Vitamin
K2 (the variation that is synthesized by the gut).
Other than food sources, the last way to obtain Vitamin K2 for your body is
through supplements. There are a wide variety of K2 supplements on the market today.
We normally get MK-4 from animals and MK-7 from our gut bacteria (Hall, 2014). And
the supplements one will find on the shelves mainly focus on delivering MK-7.
There are natural means of producing Vitamin K2, food sources of K2, as well as
supplemental sources of Vitamin K2. With so many readily available sources, one may
think that intaking sufficient amounts of K2 will decrease their risk of osteoporosis and
cardiovascular disease. However in actuality the FDA hasnt approved any form of
vitamin K for the prevention of or treatment of osteoporosis. Thats not to say there are
no correlations between the two, but rather any found correlations havent been founded
or backed up by the FDA and therefore there is no daily recommended value for Vitamin
K2.
All in all, there have been no studies to say ingesting Vitamin K2 will show
adverse health effects. So even though the FDA has not approved K2 to be a
preventative supplement, it doesnt hurt to add more foods rich in Vitamin K2 in your
diet.
Conclusion
As discussed throughout this paper, vitamin K2 can provide numerous health
benefits when it is incorporated in the diet. It has been shown to improve both heart and
bone health greatly, and as research continues on the molecule, even more benefits
may arise. In order to receive all of the benefits that vitamin K2 has to offer, it is
essential to understand some of the chemistry of it and how to properly incorporate it
into the diet.
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Works Cited
Allison AC. The possible role of vitamin K deficiency in the pathogenesis of Alzheimers
disease and in augmenting brain damage associated with cardiovascular
disease. Med
Hypotheses. 2001 Aug;57(2):151-5.
Aoki, C., Miura, M., Shiraki, M., Shiraki, Y. (2000). Vitamin K2 (menatrenone) effectively
prevents fractures and sustains lumbar bone mineral density in osteoporosis.
Journal of Bone and Mineral Research, 15, 515-521. Doi:
10.1359/jbmr.2000.15.3.515.
Aoshima, M. (2003). Apoptosis induction of vitamin K2 in lung carcinoma cell
lines: the possibility of vitamin K2 therapy for lung cancer.
PubMed.gov.
Bentley, Ronald & Meganathan, R. (1982). Biosynthesis of Vitamin K (Menaquinone) in
Bacteria. American Society for Microbiology, Vol. 46, 241-280.
Beulens, J.W.J., Booth, S.L., van den Heuvel, E.G.H.M., Stoecklin, E., Baka, A.,
Vermeer, C.
(2013). The role of menaquinones (vitamin K2) in human health. British Journal
of Nutrition, 110, 13571368. doi:10.1017/S0007114513001013.
Booth, Sarah L., Boyaval, Patrick, Karl, Philip J., & Walther, Barbara (2013).
Menaquinones, Bacteria, and the Food Supply: The Relevance of Dairy and
Fermented Food Products to Vitamin K Requirements. Advances in Nutrition,
Vol. 4, 463-473.
Chierici, E., Di Rienzo, T.A., Di Stasio, E., Flex, A., Flore, R., Gasbarrini, A., Gerardino,
L., Lanti, A., Lupascu, A., Ponziani, F.R., Santoliquido, A., Santoro, L., Tondi, P.,
Zocco, M.A. (2013). Something more to say about calcium homeostasis: the role
of vitamin K2 in vascular calcification and osteoporosis. European Review for
Medical and Pharmacological Sciences, 17, 2433-2440.
Das, A., Hugenholtz, J., van Halbeek, H., & Ljungdahl, L. (1989, July). Structure and
Function of a Menaquinone Involved in Electron Transport in Membranes of
Clostridium thermoautotrophicum and Clostridium thermoaceticum. Retrieved
February 10, 2016, from http://jb.asm.org/content/171/11/5823.full.pdf

Geleijnse, Johanna M. "Journal of Nutrition." Dietary Intake of Menaquinone Is

Associated with
 10

a Reduced Risk of Coronary Heart Disease: The Rotterdam Study. The Journal
of Nutrition, 1 Nov. 2004. Retrieve February 14, 2016, from
http://jn.nutrition.org/content/134/11/3100.long
Hall, Harriet. (2014). K2: The Vitamin not the Mountain. Science-Based Medicine.
Retrieved
from https://www.sciencebasedmedicine.org/k2-the-vitamin-not-the-mountain/
Iwamoto, J., Sato, Y., Takeda, T. (2004). Effects of vitamin K2 on osteoporosis. Current
Pharmaceutical Design, 10, 2557-2576. doi:10.2174/1381612043383782.
Jacob, A. (2013). Vitamin K2 A little-known nutrient can make a big difference in heart
and
bone health. Todays Dietician, 15, 54. Retrieved from
http://www.todaysdietitian.com/newarchives/060113p54.shtml
Kresser, Chris. (2008). Vitamin K2: The Missing Nutrient. Retrieved from
http://chriskresser.com/vitamin-k2-the-missing-nutrient/
Leech, J. (2015, July). Vitamin K2: Everything You Need to Know. Retrieved from
http://authoritynutrition.com/vitamin-k2/
Lian, J., Hauschka, P., & Gallop, P. (1978, October). Properties and Biosynthesis of a
Vitamin
K-dependent Calcium Binding Protein in Bone. Retrieved February 10, 2016,
from
http://www.ncbi.nlm.nih.gov/pubmed/700171
Linseisen, J. (2008). Dietary intake of vitamin K and risk of prostate cancer in
the Heidelberg cohort of the European Prospective Investigation into Cancer
and Nutrition. PubMed.gov.
Mercola, J. (2012). What you need to know about vitamin K2, D and calcium.
Mercola.com.
Retrieved from
http://articles.mercola.com/sites/articles/archive/2012/12/16/vitamin-
k2.aspx
National Academies Press. (2002). Dietary Reference Intakes for Vitamin A, Vitamin K,
Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybednum,
Nickel,
Silicon, Vanadium, and Zinc. Retrieved from
http://www.nap.edu/read/10026/chapter/1
Ramirez, J. (2014, September). The Surprising Longevity Benefits of Vitamin K.
Life Extension.
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Schurgers, L. J., Spronk, H.M.H., Skepper, J. N., Hackeng, T.M. Shanahan, C.M.,
Vermeer, C.,
Weissberg, P.L., & Proudfoot, D. (September 2007). Post-translational
Modifications
Regulate Matrix Gla Protein Function: Importance for Inhibition of Vascular
Smooth
Muscle Cell Calcification. Retrieved February 10, 2016, from
http://onlinelibrary.wiley.com/doi/10.1111/j.1538-7836.2007.02758.x/full