Methylation is one of the most important biological processes in the human body. It is foundational for our mental, physical, and emotional health, affecting neurotransmitter production, cell function, DNA repair, gene expression, detoxification, and much more.

The problem, however, is that our methylation ability is vulnerable to being impaired, leading to health issues.

This can happen due to poor diet, nutritional imbalances, environmental toxins, oxidative stress, and certain genetic mutations (e.g. MTHFR).

When this happens, we can experience negative symptoms such as mood swings, depression, anxiety, food sensitivities, fatigue, sleep issues, hormone dysregulation, brain fog, and much more.

The good news is that there is a lot we can do to improve our methylation ability.

However, in order to understand how to improve methylation, we must first define it.

Table of Contents

• What is Methylation?
• Why is Methylation Important?
• Nutrients Involved in Methylation
• The Methylation Cycle
• Causes of Poor Methylation
• How to Improve Methylation
• Beware of Folic Acid
• Dietary Sources of Folate
• How to Test Methylation Status
• Conclusion

What is Methylation?

When we consume food, our body takes the nutrients from that food puts them through a series of biochemical processes to transform them into new, more useable forms of those nutrients so that they can be used to activate a host of cellular processes critical to survival.

This process of converting raw materials into more usable forms that the body can use is called methylation.

In order to change nutrients into different forms, our body uses these things called methyl groups, which are molecules that consist of one carbon atom bound to three hydrogen atoms (CH3).

Methylation, in scientific terms, is the process of transferring these methyl groups from one molecule to another [1][2][3][4][5].

Our body is constantly attaching and detaching these methyl groups to various molecules, proteins, and DNA in order to change their structure and/or function.

Methylation is responsible for a vast range of functions including:

• DNA repair – Modifying DNA structure and influencing gene expression
• Detoxification – Enabling the liver to process and eliminate toxins
• Gene expression – Supporting cellular energy metabolism
• Energy production – Influencing immune cell activity and responses
• Neurotransmitter production – Enabling the synthesis of dopamine, serotonin, etc.
• Hormone regulation – Regulating hormone production, release, and metabolism
• Homocysteine Metabolism: Maintaining healthy homocysteine levels [2][5][7]

When methylation is functioning optimally, it is supporting our heart health, our mental health, our energy production, detoxification systems, our hormones, and much more.

Why is Methylation Important?

Methylation is happening every second in our body, affecting nearly every aspect of our health. Given this fact, maintaining healthy methylation is vital if we want to experience optimal mental and physical health.

There are two different scenarios we can find ourselves in when methylation is out of balance:

1. Undermethylation
2. Overmethylation

Undermethylation

Undermethylation can happen when our body is unable to generate enough methyl donors, such as folate, in order to methylate properly or if the body is being bombarded with environmental stressors that are draining our body of its methyl groups. Both of these situations leave our body struggling to keep up with its methylation demands.

People who are under methylating will typically experience things like:

• Poor mental health (depression, anxiety, OCD, etc.)
• Negative thought patterns
• Nervous system issues
• Addiction
• Gut health issues
• Fatigue
• Thyroid health issues
• Trouble sleeping
• Poor exercise capacity
• Histamine intolerance
• Heart problems
• Food sensitivities
• Increased risk of fatty liver disease
• Elevated homocysteine [1][5]

Overmethylation

Overmethylation can happen if we are lacking certain nutrients that regulate our methylation (such as glycine), preventing our body from clearing excess methyl groups, or if we are consuming too many methyl donors, like vitamin B12 or folate, leaving us with an excess and kicking our methylation into overdrive. Both of these scenarios leave us with methylation that is too high.

People experiencing symptoms of overmethylation typically report the following:

• Difficult time focusing
• Easily distracted
• Substance abuse
• Impulsivity
• ADHD
• Anger/irritability
• Anxiety/panic attacks
• Decreased ability to break established habits
• Insomnia [1]

How do we balance methylation?

The key to balancing the methylation system is to consume plenty of nutrients that support our ability to methylate while also consuming adequate amounts of nutrients that act as a buffer against excess methyl groups.

This way we supply the body with what it needs to methylate while also giving it the resources it needs to metabolize excess methyl groups, preventing both undermethylation and overmethylation.

This leads to the next question: Which nutrients are important for methylation?

Nutrients Involved in Methylation

Proper methylation depends on an adequate supply of certain micronutrients that act as methyl donors and cofactors.

The following are the key nutrients involved in methylation:

• Folate (B9)
• Riboflavin (B2)
• Vitamin B6
• Vitamin B12
• Methionine
• S-adenosylmethionine (SAM)
• Betaine (trimethylglycine)
• Choline
• Glycine
• Homocysteine

Folate 

Folate is a water soluble B vitamin (B9) that cannot be synthesized endogenously and must be consumed through diet and supplementation.

The primary function of folate is the transfer of methyl groups to allow homocysteine to be recycled back to methionine, a process called remethylation.

There are two separate remethylation pathways, one uses folate while the other uses betaine.

Folate is essential for DNA synthesis, cellular growth, protein metabolism, and neurotransmitter synthesis [1].

Riboflavin

Riboflavin, vitamin B2, is responsible for supporting vitamin B12 and folate in recycling homocysteine back into methionine [1].

Riboflavin is also used by the MTHFR enzyme to produce methyl folate.

Vitamin B6

Like riboflavin, vitamin B6 is also responsible for supporting vitamin B12 and folate in recycling homocysteine back into methionine [1].

Vitamin B12

Vitamin B12 is used as a cofactor to an enzyme called Methionine Synthase (MS) that converts homocysteine back to methionine.

Methionine

Methionine is an amino acid that is converted into the universal methyl donor called S-adenosylmethionine (SAM). It is always the starting point for providing methyl groups.

Once used, it becomes S-adenosylhomocysteine (SAH), which is then transformed into homocysteine. It is at this point when the body must recycle homocysteine back into methionine so that the process of methylation can start again [1][8].

S-adenosylmethionine (SAM)

The universal methyl donor is S-adenosylmethionine (SAM). SAM is a derivative of adenosine and methionine, which is the most important substrate in methylation process [5].

SAM is necessary for DNA and protein regulation that are important for proper prenatal development and neonatal development.

After delivering its methyl group to target sites like DNA, SAM is transformed into S-adenosylhomocysteine (SAH), which is then transformed into homocysteine and adenosine via the enzyme adenosylhomocysteinase [5].

Betaine

Also known as trimethylglycine, betaine is responsible for the second remethylation pathway, where it converts homocysteine back into methionine.

Betaine is mainly derived from choline, although it can be acquired directly through diet and supplementation as well.

Choline

In the methylation system, choline is converted into betaine where it is used to recycle homocysteine to methionine.

If choline and betaine levels are low, folate and B12 levels will need to be higher to compensate for lowered remethylation through the betaine pathway [8].

Glycine

Glycine is an amino acid that acts as a buffer for excess methyl groups. If we are consuming or producing more methyl groups than our methylation system demands, the body uses glycine to remove the excess to avoid overmethylation.

Homocysteine

Homocysteine is an intermediate amino acid in the production of methionine, which is needed for methylation.

Once methionine donates its methyl group, it is ultimately converted into homocysteine.

It is crucial that this homocysteine be recycled back to methionine so methylation can continue or else homocysteine can accumulate to high levels, leading to symptoms of undermethylation and increases in risk of certain chronic diseases like cardiovascular disease.

Other nutrients

Other important nutrients that are required to support methylation include:

• Thiamine (B1)
• Niacin (B3)
• Iron
• Phosphorus
• Sulfur
• Magnesium
• Potassium
• Zinc
• Vitamin A [1]

The Methylation Cycle

There are two primary cycles that make up the methylation cycle:

1. The Folate Cycle
2. The One-Carbon Group Cycle

The overall purpose of these cycles is to generate methyl groups that will be donated to certain molecules in order to make them usable inside the body [5].

The Folate Cycle

The ultimate goal of the folates cycle is to take folate that we consume from our diet and convert it into its usable form, L-5-methyltetrahydrofolate (L-5-MTHF), so that it can be used by methionine synthase (MS) to convert homocysteine back into methionine.

Depending on the type of folate we consume, this process can take several steps to complete.

For example, if we consume folate in the form of L-5-MTHF then the body doesn’t have to convert it because it’s already in its usable form.

However, if we consume folic acid, which is a synthetic, man-made chemical that doesn’t exist in nature, then it requires several steps to convert it to L-5-MTHF in order to be used by the body.

This conversion happens in the following sequence:

1. Folic acid to DHF

Folic acid is taken from the blood and converted into dihydrofolate (DHF) by dihydrofolate reductase (DHFR).

2. DHF to THF

DHF is then converted into tetrahydrofolate (THF) by the same enzyme, DHFR.

3. THF to 5-10-MTHF

THF is then converted into 5-10-methylenetetrahydrofolate (5-10-MTHF) by serine hydroxymethyl-transferase (SHMT).

4. 5-10-Methylene THF to L-5-Methyl THF

5-10-MTHF is finally converted into L-5-methyltetrahydrofolate by the infamous MTHFR enzyme, methylenetetrahydrofolate reductase [7].

The One-Carbon Cycle

The ultimate goal of the one-carbon cycle is to generate the universal methyl donor, SAM, so that it can be used for methylation processes such as neurotransmitter synthesis, DNA repair, detoxification, etc.

The One-Carbon Cycle uses the L-5-MTHF that’s generated from the folates cycle to accomplish this.

Steps:

1. Methionine to SAM

Methionine, obtained from dietary protein, is converted to S-adenosylmethionine (SAM) by the enzyme methionine adenosyltransferase.

2. SAM to SAH

SAM donates methyl groups to various targets, including DNA and proteins, becoming S-adenosylhomocysteine (SAH).

3. SAH to Homocysteine

SAH is converted to homocysteine.

4. Homocysteine Back to Methionine

Homocysteine is converted back to methionine through a process called remethylation.

There are two routes available for remethylation:

1. Methionine Synthase (MS) – This pathway uses folate (5-MTHF) as the methyl donor and vitamin B12 as a cofactor to convert homocysteine back into methionine

2. Betaine-Homocysteine Methyltransferase – This pathway uses betaine, or trimethylglycine, as the methyl donor to convert homocysteine back into methionine

Alternatively, homocysteine can enter the transsulfuration pathway where it’s used to generate glutathione (GSH), which is our body’s master antioxidant.

Causes of Poor Methylation

Key Takeaways:

• Methylation can be negatively impacted by certain genetic mutations, oxidative stress, diet and lifestyle choices, and nutrient deficiencies
• The following genetic polymorphisms, or mutations, can lead to dysfunction of certain enzymes involved in methylation: MTHFR (the most well-known), SLC19A1, MTHFD1, MTRR, PEMT
• MTHFR
  • MTHFR is the gene responsible for creating the enzyme that generates methyl folate
  • Mutations in MTHFR can lead to reduced production of methyl folate
  • Two primary mutations: C677T & A1298C
  • These mutations can lead to a 30 – 75% reduction in MTHFR activity
  • Around 25 – 30% of the global population has one or more of these mutations
  • Simply getting enough riboflavin could potentially offset any reduction in methylation due to MTHFR mutations
• SLC19A1
  • SLC19A1 is an enzyme that transports folate into cells.
  • Defects in this gene can lead to low intracellular folate.
• MTHFD1
  • MTHFD1 is an enzyme that produces methylenefolate, which is the precursor to methylfolate.
  • Defects in this enzyme can decrease the intracellular supply of methylenefolate, leading to increased risk of anemia.
• MTRR
  • Under conditions of oxidative stress, this defect can lead to increased risk of vitamin B12 deficiency
• PEMT
  • An enzyme that uses the methylation system to produce phosphatidylcholine.
  • Decreased PEMT activity leads to increased risk of fatty liver and liver damage in the context of a low-choline diet.
• These genetic mutations can have varying degrees of effects on individuals, largely influenced by the quality of their diet and lifestyle choices as well as environmental toxin exposure
• Oxidative stress resulting from a diet high in ultra-processed foods and low in antioxidants as well as environmental toxin exposure (tap water, personal care products, household cleaning products, plastics, pesticides, etc.) can lead to decreased methylation.
• Certain nutrient deficiencies can lead to decreased methylation such as: riboflavin, vitamin B6, folate, vitamin B12, and choline.

Methylation is negatively impacted by several things:

1. Certain genetic polymorphisms
2. Oxidative stress resulting from dietary and lifestyle choices and environmental exposures
3. Nutrient deficiencies

1. Genetic Polymorphisms

To preface this, just because you may have certain genetic polymorphisms that can negatively impact your methylation, this doesn’t necessarily mean that you will experience symptoms.

The MTHFR polymorphism, for example, is actually very common, but its effects on people can vary significantly. This variation is influenced by other genetic factors, dietary and lifestyle choices, and environmental exposures.

Experts on methylation like Chris Masterjohn and Ben Lynch have said the same thing. These genetic mutations may increase your risk for having poor methylation activity, but your genes do not determine your destiny, and what truly matters is how those genes are expressing themselves. Gene expression can be modulated by the way we live our lives and the foods we eat.

In addition, there are specific nutrients we can consume to offset the impact that these mutations have, which we will cover in the next section.

So, if you are someone who is actively experiencing symptoms and/or whose lab results indicates methylation issues (e.g. high homocysteine), there is no need to panic or stress as there is much we can do to improve it.

The following genetic mutations can impair our body’s ability to methylate:

• MTHFR
• SLC19A1
• MTHFD1
• MTRR
• PEMT

MTHFR

MTHFR is a huge buzzword in the health and wellness space and is probably the most highly talked about genetic mutation.

But what exactly is it and why is it relevant?

MTHFR is the gene that is responsible for providing our body with instructions for creating the enzyme 5, 10-methylenetetrahyfrofolate reductase (MTHFR), which is the enzyme that transforms 5,10-methylenetetrahydrofolate into the active form of folate, 5-methyltetrahydrofolate (5-MTHF) [5][1].

Basically, it is the final enzymatic step in our body’s process of converting dietary folate, or folic acid, into the usable form of folate that our body will use for methylation.

Genetic mutations called single nucleotide polymorphisms (SNPs) can occur in the MTHFR gene, resulting in many variants of the enzyme, reducing its ability to generate 5-MTHF [1].

There are two common MTHFR genetic mutations or SNPs that can hurt MTHFR activity:

1. C677T – People who carry this mutation will have a reduced capacity to create L-methylfolate (5-MTHF)
2. A1298C – This polymorphism reduces MTHFR enzyme activity to a lesser extent than C677T

When these mutations are present, the MTHFR enzyme activity is reduced, leading to low production of folate (5-MTHF), which leads to poor methylation.

Impact on MTHFR Activity

C677T

• Heterozygous (C/T) = 33 – 35% reduction in MTHFR activity
• Homozygous (T/T) = 70 – 75% reduction in MTHFR activity [15][19]

A1298C

• Heterozygous (A/C) = 15 – 17% reduction in MTHFR activity
• Homozygous (C/C) = 30 – 39% reduction in MTHFR activity [15][19]

C677T + A1298C

• Having one C677T allele (677 CT) + one A1298C allele (1298 AC) = 53% reduction in MTHFR activity [15]

Prevalence

The frequency of these genetic polymorphisms varies depending on geographic location and ethnicity, but according to the Genome Aggregation Database and the 1000 Genomes Project, here is the prevalence of each polymorphism:

C677T

• Global Population: 25 – 30%
• Hispanics: 47%
• Europeans: 36% heterozygous (677 CT); 13.5% homozygous (677TT)
• East Asians: 30%
• South Asians: 12%
• Africans: 9% [11][17]

A1298C

• Global Population: 25 – 29%
• South East Asians: 42%
• Europeans: 31% heterozygous (1298 AC); 11% homozygous (1298 CC)
• Hispanics: 15%
• Africans: 15% [11][17]

C677T + A1298C

• Combined two variants, both heterozygous: >20%
• Combined two variants, one homozygous + one heterozygous: 0.4% (rare)
• Combined two variants, both homozygous: None detected (most likely a lethal combination [11]

The Bottom Line

As you can see, MTHFR polymorphisms are very common. Given that fact, it’s unlikely that these mutations are a random, bad luck of the draw. They must have served us at some point in human history.

Nutrition scientist and methylation expert, Dr. Chris Masterjohn, believes that the MTHFR mutations likely had no negative impact on humans in the context of our ancestral diet due to high intake of riboflavin (vitamin B2).

Our hunting and foraging ancestors ate large amounts of animal meat, fat, and organs, a diet that provides high amounts of riboflavin.

Riboflavin is the vitamin that the MTHFR enzyme uses to in convert dietary folate into the usable form, 5-methyltetrahydrofolate (5-MTHF).

Dr. Masterjohn theorizes that MTFHR polymorphisms only decrease MTHFR activity because most people in whom the activity has been measured have inadequate levels of riboflavin [20].

His reasoning is that the MTHFR polymorphisms decrease the activity of the MTHFR enzyme by making it bind more weakly to riboflavin, and since the MTHFR enzyme needs to bind to riboflavin in order to carry out its task of generating 5-MTHF, low levels of riboflavin reduces the activity of the MTHFR enzyme even more [20].

Studies have shown that at high enough concentrations of riboflavin, enzymatic activity the MTHFR enzyme is restored to normal.

For example, supplementation with 1.6 mg of riboflavin has been shown to decrease homocysteine levels by 40% in those with the MTHFR C667T polymorphism + poor riboflavin status [22].

This means that by simply getting enough riboflavin through our diet and/or supplementation, we could potentially cancel out any decrease in MTHFR enzyme activity caused by the MTHFR polymorphisms.

In addition, nutritional researcher Dr. Alex Leaf has found that in humans with the MTHFR C667T polymorphism, all of the elevated homocysteine is concentrated in those with low levels of riboflavin [20].

SLC19A1

• SLC19A1 is an enzyme that transports folate into cells.
• Defects in this enzyme can decrease the intracellular supply of methylenetetrahydrofolate, or simply methylenefolate, which is the form of folate that prevents anemia.
• This mutation can raise one’s risk of developing anemia.
• Homozygosity for G80A allele decreases SLC19A1 enzyme activity by 50% [15].

MTHFD1

• MTHFD1 is an enzyme that produces methylenefolate, which is the precursor to methylfolate.
• Defects in this enzyme can decrease the intracellular supply of methylenefolate, leading to increased risk of anemia.
• Homozygosity for the G1958A allele decreases MTHFD1 enzyme activity by 34% [15].

*Since the two polymorphisms above impact the supply of methylenefolate, folate supplementation should include a mix of folinic acid and methylfolate since folinic acid will be more easily converted to methylenefolate.

MTRR

• MTRR is an enzyme that repairs vitamin B12 when it has been damaged by oxidative stress.
• Homozygosity for A66G or C524T lowers the activity of the enzyme by 300-400%.
• Under conditions of low oxidative stress, these mutation causes no issues.
• Under conditions of oxidative stress, these mutations increase the risk of vitamin B12 deficiency.
• These mutations do not affect nutritional requirements [15].

PEMT

• An enzyme that uses the methylation system to produce phosphatidylcholine.
• Decreased PEMT activity leads to increased risk of fatty liver and liver damage in the context of a low choline diet.
• The way to mitigate this is to consume adequate amounts of choline [15].

2. Oxidative Stress

Our bodies naturally produce molecules known as reactive oxygen species (ROS) as a result of normal biological processes such as cellular respiration, where oxygen is used to produce energy.

ROS are unstable molecules that cause damage in the body.

Our bodies have built-in antioxidant systems to counteract ROS and prevent them from causing damage.

However, when ROS production exceeds our body’s ability to neutralize them, it can lead to oxidative stress, which is a state where cellular damage occurs due to the harmful effects of ROS. This can lead to premature aging and a host of chronic diseases.

When we experience oxidative stress for too long, we can also start experiencing issues with methylation.

For example, prolonged oxidative stress will lead to increased demand for our body’s master antioxidant, glutathione. Since we need to break down homocysteine to make glutathione, this means that we will have excessive breakdown of homocysteine and less available to be recycled back into methionine by MTHFR. This leads to lower levels of our universal methyl donor, SAM [8].

That begs the question: What sorts of things lead to excessive ROS production?

Our modern environment is plagued with ROS that generate oxidative stress in the body.

Key examples are:

• An unhealthy diet
  • Ultra-processed foods
  • Hydrogenated seed oils
  • Refined sugar and flour
  • Artificial food dyes
  • Preservatives, etc.
  • Low intake of antioxidants
• Environmental toxins
  • Air pollution
  • Smoking
  • Tap water
  • Heavy metal exposure
  • Mold toxicity
  • BPA & other plastics
  • Phthalates
  • Sunscreens, lotions, soaps
  • Personal care products
  • Household cleaning products
  • Pesticides and herbicides
  • PFAS, PCBs, VOCs, etc.
  • Poor indoor air quality
• Nutrient Deficiencies
  • Vitamin B2 (riboflavin) – Low riboflavin means the MTHFR enzyme won’t be able to produce methyl folate as effectively, leading to low folate levels and high homocysteine.
  • Vitamin B6 (pyridoxine) – B6 helps in recycling homocysteine back to methionine. Low levels can cause homocysteine to accumulate in the body.
  • Vitamin B9 (folate) – Low dietary intake of folate can lead to decreased methylation and high homocysteine.
  • Vitamin B12 (cobalamin) – Low intake of B12 can reduce the conversion of homocysteine back to methionine, leading to high homocysteine.
  • Choline – Low intake of choline will cause low levels of betaine, which is needed to convert homocysteine back to methionine using the alternative remethylation pathway, betaine-homocysteine methyltransferase.

How to Improve Methylation

Since we know that methylation is negatively impacted by genetic mutations, nutrient deficiencies, and unhealthy diet and lifestyle choices, our solution will be to address each one of these.

Our overall objective will be to supplement the specific nutrients that support these genetic mutations and neutralize their impact, reduce oxidative stress by cleaning up our diet and lifestyle, and reducing our toxic burden by cleaning up our environment.

However, while the recommendations below can be helpful, it is crucial to recognize that outcomes from supplementation or lifestyle changes may vary depending on the individual.

Having said that, we can take a comprehensive approach to improve our methylation using the following strategies:

1. Nutritional Supplementation
2. Dietary Changes
3. Lifestyle Changes

1. Nutritional Supplementation:

The following nutritional supplementation guidelines were pulled together based on recommendations made by several methylation experts including Dr. Chris Masterjohn and Dr. Ben Lynch. We also performed independent research and have personally experimented with these protocols.

It’s important to note that improving methylation through supplementation requires a very personalized approach as everyone will respond differently to these nutrients. It’s best to introduce one supplement at a time and track changes before moving on to the next.

Key Takeaways:

1. Folate: 400-1000 micrograms/day of methylfolate or folinic acid
2. Betaine: 1000 mg/day of trimethylglycine (500mg twice/day)
3. Riboflavin: 3-400 mg/day (experiment with the dose)
4. Vitamin B12: Consume animal foods for vitamin B12 and/or supplement with hydroxycobalamin or methylcobalamin (no specific dosage recommended)
5. Glycine: 10 g/day (3 g/meal)
6. Creatine: 3-5 g/day of creatine monohydrate
7. Phosphatidylcholine: 1200 mg/day
8. B-complex and trace minerals: Use as recommended; make sure b-complex is methylated

Objective:

The overall objective is to have balanced methylation. We achieve this by giving the body a sufficient supply of nutrients that offer methyl groups on one hand while consuming plenty of glycine as a buffer excess methyl groups on the other.

Here is a breakdown of the steps:

1. Support folate remethylation pathway
2. Support betaine remethylation pathway to take burden off of folate remethylation pathway
3. Improve MTHFR enzyme function
4. Increase intake of cofactors necessary for utilization of folate
5. Support methyl buffer system
6. Reduce demand on the methylation cycle
7. Support Methylation With B-Complex and Trace Minerals

Support Folate Remethylation Pathway

• Supplement with methylfolate or folinic acid
• Since the MTHFR enzyme isn’t functioning properly and isn’t generating enough methyfolate, we can make up the difference by supplementing with methylfolate
• Folinic acid still needs to be turned into methyfolate by MTHFR
• This makes it less likely to cause side effects related to a sudden increase in methyl groups
• The FDA has requirements regarding how folic acid is listed on the label for dietary supplements. For these products, the amount is listed as mcg of dietary folate equivalents (DFE) and then as mcg of folic acid. The DFE measure is used because the body absorbs folic acid easier than folate. So when you look at the label, you’ll see something like this: Folate 665 mcg DFE (400 mcg Folic Acid).
• Dosage: 1000 DFE (Dietary Folate Equivalents) mcg/d

Note: You’ll notice on the supplement facts label that the amount of folate is listed as DFE (Dietary Folate Equivalents) and then again as mcg of folic acid. This does not mean that the supplement contains folic acid. The DFE measurement is used because the body absorbs folic acid better than folate. The label is showing you the equivalent amount of folic acid that you would be consuming with the given amount of methyl folate in the supplement. The FDA requires this type of labeling on all supplements containing folate or folic acid.

Support Betaine Remethylation Pathway To Reduce Demand on Folate Remethylation Pathway

• Supplement with trimethylglycine (TMG)
• TMG serves as an alternative to methylfolate
• It allows us to bypass an underperforming folate remethylation pathway and use the betaine remethylation pathway instead
• It reduces burden on the already struggling folate pathway
• Dosage: 500mg twice a day

Improve MTHFR Enzyme Function

• Supplement with riboflavin
• Riboflavin helps improve the ability of the MTHFR enzyme to generate methylfolate
• Dosage: 3-400 mg/d

Increase Intake of Cofactors Necessary For Utilization of Folate

• Supplement with B12 if deficient
• B12 is vital to use the methylfolate to convert homocysteine back to methionine via the methionine synthase enzyme (MS)
• If B12 levels are too low, the body won’t be able to use the methylfolate to convert homocysteine back to methionine, even if folate levels are optimal
• This can lead to build up of methylfolate and homocysteine
• If you are not deficient in B12, supplementation isn’t as important

Support Methyl Buffer System

• Supplement with glycine
• Glycine acts as a buffer to excess methyl groups, storing them and retrieving them when needed
• This prevents a buildup of methyl groups which can lead to over methylation which has negative consequences
• It serves to maintain stable methylation activity, preventing swings in things like energy, mood, etc.
• Dosage: 3 grams per meal (3-10g/day total)

Reduce Demand on the Methylation Cycle

• Supplement with creatine monohydrate
• Creatine synthesis is estimated to consume around 40-45% of the body’s methyl groups, which is a significant amount [13]
• By supplementing with creatine, we can reduce the body’s reliance on endogenous creatine synthesis, thereby easing demand on methyl groups
• Dosage: 3-5 g/d
• Supplement with phosphatidylcholine or choline
• Phosphatidylcholine synthesis consumes another 30-45% of methyl groups [14]
• Best source is alpha-GPC as it contains 40% choline
• Other sources: CDP choline (aka citicoline), choline bitartrate, phosphatidylcholine
• Best to consume a mixture of phosphatidylcholine and choline
• Dosage: 1000 mg/d (8 eggs equivalent) or use Chris Masterjohn’s Choline Calculator for a more accurate recommendation based on your genetic testing – How Much Choline Should I Eat?
• Choline-rich foods: liver, eggs, lecithin, salmon
• Use Chris Masterjohn’s Choline Food Database – The Choline Database |

Support Methylation With B-Complex and Trace Minerals

• While the nutrients listed above are required to improve methylation in the context of reduced MTHFR activity, many other nutrients are needed in the background to keep methylation running smoothly.
• These include a broad spectrum of B-vitamins along with a range of trace minerals including:
  • Vitamin B1 (thiamin)
  • Vitamin B3 (niacin)
  • Vitamin B6 (pyridoxine)
  • Vitamin A
  • Iron, phosphorus, sulfur, magnesium, potassium, manganese, and zinc
• A diet emphasizing animal foods like red meat, pork, chicken, eggs, fish, and dairy should provide plenty of these nutrients
• However, supplementation may be needed in some cases, and it doesn’t always hurt to experiment and see what works
• If supplementing, the best thing to do is purchase a methylated B-complex supplement along with a trace mineral supplement

2. Dietary Changes

In order to minimize oxidative stress, we must consume a diet that focuses on eliminating inflammatory foods, including plenty of nutrients that support methylation, including antioxidants that combat oxidative stress, and eliminating pesticides to reduce toxic burden.

To achieve this, follow these tips:

1. Eliminate ultra-processed foods
   • Anything containing: hydrogenated seed oils, refined flour and sugar, pasteurized and homogenized dairy, high fructose corn syrup, synthetic food dyes, artificial flavors, preservatives, etc.
2. Eliminate gluten from your diet.
3. Reduce or eliminate commercial dairy
   • Opt for raw goat and cow’s milk instead
4. Eat a whole foods diet with an emphasis on animal foods (red meat, chicken, fish, eggs, dairy, etc.)
5. Eat nose-to-tail (organs, bones, cartilage, fat, etc.)
   • Liver, kidney, heart, spleen, etc.
   • Bone broth
   • Suet, tallow, lard, etc.
6. Consume fruits and vegetables while being mindful of any food sensitivities
7. Consume plenty of antioxidants like vitamin a, vitamin c, vitamin e, polyphenols, selenium, manganese, zinc, Co. Q-10, and glutathione
   • Consume things like citrus fruits, berries, green and black tea, matcha, liver, heart, eggs, and seafood
   • Things like glutathione and vitamin c can be supplemented as well (liposomal versions are the most effective)
8. Eat organic as much as possible to avoid pesticides, like glyphosate, that poison our cells, degrade our gut barrier, and damage our microbiome

3. Lifestyle Changes:

Lifestyle changes should consist of reducing environmental toxins, reducing habits that cause oxidative stress in the body, and supporting our body so it can combat oxidative stress and eliminate toxins more effectively.

Key changes should include:

• Eat organic food as much as possible
• Choose grass-fed beef, free-range, hormone-free, and antibiotic-free meats and eggs
• Drink filtered water or spring water only
  • Remineralize with trace minerals if filtering with an intense filter
  • Consume plenty of remineralized water daily
• Avoid drinking tap water
• Buy toxin-free cosmetics that don’t include typical endocrine disruptors such as parabens, benzophenones, bisphenols, and phthalates
  • Use the Environmental Working Group’s (EWG) website to find safer, less toxic brands
• Buy non-toxic household cleaning supplies (dish detergent, laundry detergent, kitchen and bathroom cleaner, etc.)
  • Use the Environmental Working Group’s (EWG) website to find safer, less toxic brands
• Avoid commercial sunscreens
  • Use the Environmental Working Group’s (EWG) website to find safer, less toxic brands
• Avoid cooking, drinking, storing, and heating food or drinks in plastic containers
• Avoid plastics (receipts, bottled water, cling film, plastic tupperware, etc.)
• Use an air purifier in your home and office
• Avoid using over-the-counter medications such as NSAIDS, antiacids, etc. as they can damage the gut lining, impair our absorption of nutrients, and add to our toxic load
• Stop smoking and vaping
• Limit alcohol
• Engage in activities that promote sweating such as sauna, exercise, sports, etc.
• Get plenty of high quality sleep each night
  • Shoot for 7-9 hours
  • Eliminate artificial light after sunset
  • Cool your home at night to help prepare the body for sleep (63-70 degrees Fahrenheit)
• Exercise regularly
  • Use a combination of resistance training, moderate intensity aerobic conditioning, walking, and stretching
  • Lift weights at least 2-3 times per week
  • Add in some moderate-intensity aerobic exercise like running, biking, swimming, etc.
  • Walk daily
  • Include some time each week for stretching and/or yoga
• Get adequate amounts of sunlight each day
  • Get morning sunlight for at least 10 minutes
  • Get midday/afternoon sunlight for vitamin D production
  • Sunlight is vital for immune system function, dopamine production, and hormone regulation

Other considerations:

• Remove carpets from your home and replace with low VOC wood or tile flooring
• Filter chlorine from your drinking water, shower, and bath
  • You can purchase a whole-home water filtration system if desired
• Remove mercury amalgams and root canals with the assistance of a trained biological dentist
• To promote detoxification, in addition to sweating, you can supplement with things that help pull toxins out of the body such as:
  • Liposomal glutathione
  • N-Acetyl Cysteine (NAC)
  • Binders like activated charcoal, zeolite/bentonite clay, etc.
  • Humic and fulvic acids
  • Immunoglobulins (IgG)

Beware of Folic Acid

Folic acid is a synthetic, man-made version of natural folate that is found in food.

It’s the version of folate you’ll likely see in most supplement formulations, especially prenatal multivitamins. It’s added to fortified foods like cereals, bread, flour, pasta, and rice.

It’s also the version of folate recommended by doctors for pregnant women to prevent neural tube defects.

However, consuming folic acid can cause negative effects in the body. It can negatively affect enzyme function in the folate pathway, may decrease vitamin B12 absorption in the small intestine, and ironically even potentially cause a folate deficiency.

But why do we even have folic acid in the first place, and why is it recommended by the medical community over natural versions of folate?

Before we answer that, let’s compare the different types of folate.

Types of folate:

1. L-Methylfolate: A general term for any type of folate that is methylated, such as L-5-MTHF.
2. L-5-Methyltetrahydrofolate (L-5-MTHF): This is the body’s most usable form of folate. It is the end-product of the folate pathway that supports methylation and the production of SAMe. It is also referred to as just 5-methyltetrahydrofolate (5-MTHF).
3. 5-10-Methylenetetrahydrofolate (5,10-MTHF): 5,10-MTHF is a substrate in the folate metabolic pathway. It is a crucial intermediate in the metabolism of folate, and the enzyme 5,10 methylenetetrahydrofolate reductase (MTHFR) converts it to 5-methyltetrahydrofolate (L-5-MTHFR), the main form of folate in the body.
4. Tetrahydrofolate and Dihydrofolate: These are reduced forms of folate that are found naturally in foods. Our body can easily convert these into the active form, 5-MTHF.
5. Folinic Acid: Folinic acid (not FOLIC acid) is a natural and active form of folate. It often used by the body for DNA and RNA synthesis. It is also used to treat folate deficiency that causes anemia. In the body it can either be used in its current form or converted into 5-MTHF. It’s a great alternative to 5-MTHF for individuals who are sensitive to methylated folate.
6. Folic Acid: This is the man-made, synthetic form of folate that is used to fortify foods and is commonly found in mainstream multivitamin supplements. Folic acid has no biological activity unless it is converted into folates.

Side Effects of Folic Acid

1. It does not metabolize well – The liver can only process a certain amount of folic acid, at which point any excess begins to accumulate in the blood. This is a bad thing since this accumulated unmetabolized folic acid can lead to various health issues. This does not happen with natural forms of folate [23][24].
2. It can result in a functional folate deficiency – High folic acid intake can impair the body’s ability to convert it to other active forms of folate, such as tetrahydrofolate. This leads to accumulation of UMFA in the blood. This UMFA can impair folate metabolism, leading to a state of high blood folic acid levels but impaired utilization by the body, a situation referred to as a “functional folate deficiency” [25].
3. It can mask a folate deficiency – If you’re getting lab work done, you’re folate levels might look normal even though you could be suffering from a functional deficiency. This is because most lab tests measure folate levels in the blood, not the cells.
4. It can cause adverse health effects – High blood concentrations of folic acid can lead to a number of issues including impaired cognitive development and lowered IQ in children, immune system dysregulation, cognitive impairment in adults, and increased colorectal cancer risk [26].
5. It can reduce the bioavailability of methyl folate – Folic acid has a higher affinity for folate receptors, which are important for transporting folate into cells. This means that folic acid may be preferentially transported into cells, potentially displacing true folate. Transport of methyl folate into the brain is carried out by the folate receptor, for example. So folic acid in the blood might inhibit the transport of methyl folate into the brain. In addition, transport of unmetabolized folic acid into cells can occur via the folate receptor, which means the cells are being filled with a substance the body cannot utilize [27].
6. It inhibits the MTHFR enzyme – High folic acid intake reduces MTHFR enzyme activity, creating a pseudo-MTHFR deficiency (i.e. reduced methylation capacity). This deficiency can have serious health implications like potential liver damage, increased risk of neural tube defects during pregnancy, and increased risk of certain cancers [28][29].
7. It reduces DHFR enzymatic activity – DHFR converts folic acid into its active form, tetrahydrofolate (THF). DHFR is also involved in generating biopterin, which is a coenzyme that is essential for dopamine and serotonin production as well as nitric oxide production. Folic acid competes for DHFR binding sites, leading to low biopterin levels. This can lead to impaired brain chemistry and cardiovascular issues [30][31][32].
8. High folic acid + B12 deficiency can cause adverse effects – Low vitamin B12 status is very common among older adults. Low B12 status combined with high serum folic acid levels is associated with anemia and cognitive impairment in the elderly, after controlling for factors such as demographic characteristics, cancer, smoking, alcohol intake, etc. [34][35].

Dietary Sources of Folate

The best dietary sources of folate are:

• Liver (high bioavailability)
• Egg yolks (high bioavailability)
• Dark green leafy vegetables (spinach, mustard greens, turnips, lettuce, cabbage, etc.)
• Asparagus
• Okra
• Brussels Sprouts
• Avocado
• Broccoli
• Oranges
• Beans
• Papaya
• Bananas
• Grapes
• Strawberries
• Walnuts
• Peanuts

How to Test Methylation Status

There are several ways to test your methylation status:

1. Genetic testing
2. Measure homocysteine
3. Experiment with supplementation

Genetic Testing

Tests: StrateGene, Genova Methylation Panel

Genetic testing can tell you whether or not you have certain genetic polymorphisms like MTHFR or SLC19A1, but they don’t always guarantee that you are suffering from symptoms of poor methylation.

Regardless, it’s good information to have and it’s a good place to start.

There are several companies that offer genetic testing for methylation-related genetic mutations.

One that comes highly recommended is StrateGene.

It tests for over 100 genetic variations over 9 different pathways.

In addition, you can upload your raw data file from gene sequencing companies like 23andMe or Ancestry to Dr. Chris Masterjohn’s Choline Calculator for free where it will show you which genetic polymorphisms you have.

Measure Homocysteine

Tests: Genova Methylation Panel, Quest

Elevated homocysteine is the most standard biomarker for suboptimal methylation. Several companies offer blood tests that check homocysteine levels: Quest, and the Genova Methylation Panel.

The optimal range is between 6 and 9 [18].

In addition to homocysteine, several other lab markers are important for gaining insight into your methylation status:

• SAM-to-SAH ratio
• Plasma or serum folate
• Plasma methionine, glycine, and sarcosine

All of these markers are found on the Genova Methylation Panel.

Experiment With Supplementation

The third way to test for poor methylation is to simply add in methylation supplements like folate, betaine, riboflavin, etc. and evaluate how you respond.

This was my personal approach. I found it was cheaper and quicker to order the vitamins and simply try one at a time while evaluating whether or not I noticed a difference.

It also seems to be the most reliable way to know that your methylation is suboptimal since responding positively to supplementation leaves little room for doubt while genetic testing and even lab testing may not necessarily mean you are experiencing symptoms.

In my personal case, I noticed that riboflavin supplementation (200 mg/d) made my mood and cognition noticeably worse, while B-complex and methylfolate significantly improved my mood, energy, drive, and resilience to stress.

Conclusion

Now we know how important methylation is for our mental and physical health as well as how easy it can be for our ability to methylate to get thrown off by diet, lifestyle, and certain genetic polymorphisms.

The strategies mentioned in this article have been used be me personally and have greatly improved my quality of life, and I hope it can do the same for you.