Why Cannabis Grown with the Addition of LAB Might Produce a Superior High

As an organic gardener, and a guy who generally can’t resist reading into why things happen, I’d started using LABS (Lactic Acid Bacteria Serum) in the garden soon after I began learning of its benefits. When greenhouse tomatoes were grown with the addition of a blend of humic fertilizer, produced from vermicompost, and bio-fertilizer, containing Lactobacillus casei, Lactobacillus lactis, Phodopseudomonas palistris, Sаccharomices cerevisiae that the total tomato yield was increased by 19 and 21% after soil application and by 13 and 14% after foliar application of bio-fertilizer and humic fertilizer, respectively.1  Lactic Bacteria have uses beyond just that of a bio-fertilizer though, it can even exhibit antifungal activity.2  I’d first learned about the benefits of lactic bacteria though, through research into EM1 (Effective Microorganisms). ‘The Concepts and Theories of Effective Microorganisms‘ by T. Higa and G. N. Wididana, introduced the concept of Effective Microorganisms (EM / EM1), which described an inoculant of multiple species of beneficial microorganisms, meant to provide a wide variety of uses, including ‘suppression of plant pathogens and diseases, conservation of energy in plants, solubilization of soil minerals, and soil microbial-ecological balance, photosynthetic efficiency, and biological nitrogen fixation’.3 The blend of microorganisms originally described were a mix of lactic acid bacteria, purple bacteria, yeast, actinomycetes, and fermenting fungi . T. Higa went on to create the brand EM™ Technology, and later revised his mixture, to only include lactic acid bacteria, purple bacteria, and yeast.
1 – ‘The Effects of Humic and Bio-Fertilizers on Growth and Yield of Greenhouse Tomatoes’, I. Tringovska
2 – ‘Broad and complex antifungal activity among environmental isolates of lactic acid bacteria’, Jesper Magnusson, Katrin Ström, Stefan Roos, Jörgen Sjögren, Johan Schnürer
3 – ‘The Concepts and Theories of Effective Microorganisms‘ T. Higa and G. N. Wididana, Page 1

We can create something in the same vein as EM1, minus the purple bacteria, by fermenting our own LABS. The process is quite simple, and requires only a few basic ingredients, fresh rice, water, milk, molasses, a jar, and a one way fermentation valve. You soak the rice in the jar, shaking vigorously, then straining the rice, and keeping the now milky water. This water is now a carbohydrate rich solution. Now cover the jar loosely, so bugs don’t get in, and let stand in the back of your cupboard for a few weeks. At the point it forms a skin on the top, and you can see three layers it’s ready. Use a siphon to recover the middle layer. Take that extracted solution and mix it with 10 parts milk, then seal the jar with a one way fermentation valve. Around 7 – 8 days of fermentation, the dropping ph will cause the excess milk fats and carbohydrates to gather into a curd, a probiotic whey produced as a byproduct of the process. This curd can be blended in with a SST, thrown in compost, or fed to a pet as long as the process was done successfully, but what we really want is the water below the whey. Strain this solution and add equal parts molasses (1L of Serum:1L of Molasses), to stabilize the serum.  A more in depth guide to LABS, with pitctures can be found at http://theunconventionalfarmer.com/recipes/lactobacillus-serum/ or for a more scholarly approach from the University of Hawaii at http://www.ctahr.hawaii.edu/oc/freepubs/pdf/sa-8.pdf

Hypothesis: Potential method of actions for the increased trichome to calyx ratio, as well as the increased terpene production observed after use of homemade Lactic Acid Bacteria Serum

My current hypothesis is that homemade LABS contains some quantity of the short-chain fatty acid hexanoate, as a byproduct of microbial esterification during the fermentation of ruminant milk fats .

To follow this I’m assuming you already know some basics regarding fatty acid biosynthesis, and its relation to both cannabinoid, and terpene biosynthesis. These are phytochemical reactions that occurs in plant tissue that eventually end in the cannabinoids and smells we all know and love. If you aren’t yet familiar, here’s an incredibly simplified crash course.

Below is a basic look at cannabinoid biosynthesis. Think of this as the steps the plant has to perform to create cannabinoids and terpenes. (Cannabinoids are terpenophenolic compounds, meaning they are composed of part terpene, and part phenol group.)

This first tree starts from Geranyl Pyrophosphate & Olivetolic Acid (The olivetolic acid production of a plant is what increases when subjected to UV-B spectrum of light, this is what is being talked about when lighting technology mention more trichome through UV spectrum) and ends in the acidic cannabinoids (Acidic cannabinoids being what is found in fresh material, with the additional carboxyl (-COOH ) group. Getting rid of this -COOH group is why we cure our smoke, or decarb our hash. )

This is a great place to start, but we need to go a little more in depth. So below, is a much more robust view of the biosynthesis occurring.

Hexanoyl-CoA, which is derived from the short-chain fatty acid hexanoate, is used as a primer for a polyketide synthase (PKS) enzyme that forms olivetolic acid (OA).
Hexonate is formed via esterification of hexonic acid. So in this case we would see a proposed increase via application of Hexonate (Hexonic acid) stimulating hexonyl-CoA, thereby stimulating olivetolic acid production.

Hexanoic acid (caproic acid), is the carboxylic acid derived from hexane with the general formula C5H11COOH. It is a fatty acid found naturally in various animal fats and oils, and is one of the chemicals that give the decomposing fleshy seed coat of the ginkgo its characteristic unpleasant odor.

Hexanoic acid (6:0) comprises 1-2% of the total fatty acids in ruminant milk triacylglycerols, where most of it is esterified to position 3 of the triacyl-sn-glycerols. It is also found as a minor component of certain seed oils rich in medium-chain saturated fatty acids..
Medium-chain fatty acids, such as octanoic (8:0), decanoic (10:0) and dodecanoic (12:0), are found in esterified form in most milk fats, including those of non-ruminants, though usually as minor components, but not elsewhere in animal tissues in significant amounts. They are never detected in membrane lipids, for example. They are absent from most vegetable fats, but with important exceptions. Thus, they are major components of such seed oils as coconut oil, palm kernel oil and Cuphea species.
Odd-chain fatty acids from 13:0 to 19:0 are found in esterified form in the lipids of many bacterial species, and they can usually be detected at trace levels in most animal tissues, presumably having been taken up as part of the food chain. In particular, they occur in appreciable amounts (5% or more) in the tissues of ruminant animals.

Ruminant milk triacylglycerols, are milk fats, of a sort.
“Ruminant milk fat is of unique composition among terrestrial mammals, due to its great diversity of component fatty acids. The diversity arises from the effects of ruminal biohydrogenation on dietary unsaturated fatty acids and the range of fatty acids synthesized de novo in the mammary gland.”

This hexonic acid is our potential catalyst for increased fatty acid biosynthesis, as well as increasing a variety of plant defense systems that can also have an effect on trichome production. 1 2

1 – Priming of plant resistance by natural compounds. Hexanoic acid as a model
The data presented in this review reflect the novel contributions made from studying these natural plant inducers, with special emphasis placed on hexanoic acid (Hx), proposed herein as a model tool for this research field. Hx is a potent natural priming agent of proven efficiency in a wide range of host plants and pathogens. It can early activate broad-spectrum defenses by inducing callose deposition and the salicylic acid (SA) and jasmonic acid (JA) pathways. Later it can prime pathogen-specific responses according to the pathogen’s lifestyle. Interestingly, Hx primes redox-related genes to produce an anti-oxidant protective effect, which might be critical for limiting the infection of necrotrophs. Our Hx-IR findings also strongly suggest that it is an attractive tool for the molecular characterization of the plant alarmed state, with the added advantage of it being a natural compound.

2 – The hexanoyl-CoA precursor for cannabinoid biosynthesis is formed by an acyl-activating enzyme in Cannabis sativa trichomes
The psychoactive and analgesic cannabinoids (e.g. D9 -tetrahydrocannabinol (THC)) in Cannabis sativa are formed from the short-chain fatty acyl-coenzyme A (CoA) precursor hexanoyl-CoA. Cannabinoids are synthesized in glandular trichomes present mainly on female flowers. We quantified hexanoyl-CoA using LC-MS/MS and found levels of 15.5 pmol g)1 fresh weight in female hemp flowers with lower amounts in leaves, stems and roots. This pattern parallels the accumulation of the end-product cannabinoid, cannabidiolic acid (CBDA). To search for the acyl-activating enzyme (AAE) that synthesizes hexanoyl-CoA from hexanoate, we analyzed the transcriptome of isolated glandular trichomes. We identified 11 unigenes that encoded putative AAEs including CsAAE1, which shows high transcript abundance in glandular trichomes. In vitro assays showed that recombinant CsAAE1 activates hexanoate and other short- and medium-chained fatty acids. This activity and the trichome-specific expression of CsAAE1 suggest that it is the hexanoyl-CoA synthetase that supplies the cannabinoid pathway. CsAAE3 encodes a peroxisomal enzyme that activates a variety of fatty acid substrates including hexanoate. Although phylogenetic analysis showed that CsAAE1 groups with peroxisomal AAEs, it lacked a peroxisome targeting sequence 1 (PTS1) and localized to the cytoplasm. We suggest that CsAAE1 may have been recruited to the cannabinoid pathway through the loss of its PTS1, thereby redirecting it to the cytoplasm. To probe the origin of hexanoate, we analyzed the trichome expressed sequence tag (EST) dataset for enzymes of fatty acid metabolism. The high abundance of transcripts that encode desaturases and a lipoxygenase suggests that hexanoate may be formed through a pathway that involves the oxygenation and breakdown of unsaturated fatty acids.