Bacteria and fungi are living organisms that produce specific enzymes to break down compounds so that they can be ingested through the bacterial cell wall. Each type of enzyme is highly specific, targeting only one type of molecule. To completely degrade a mixed waste, selected groups of complementary enzymes must be produced. Most strains of bacteria are capable of producing 2 or more enzyme types. A selection process determines which enzymes are produced and in what strength. Although all enzymes are proteins, not all proteins are enzymes. File updated 10/12/2004

BASIC CLASSES OF ENZYMES:

• Oxidoreductases - catalyze oxidations or reductions. Examples: dehydrogenases, oxidases, and peroxidases.
• Transferases - catalyze the transfer of a group from one molecule to another. Examples: Phosphatases, transaminases, and transmethylases.
• Hydrolases - catalyze hydrolysis reactions. Examples: digestive enzymes such as sucrase, amylase, maltase, and lactase.
• Lyases - catalyze the removal of groups in non-aqueous media. An example would be the decarboxylases.
• Isomerases - catalyze the isomerization of molecules. Examples: racemases, and cis-trans isomerases.
• Ligases (aka synthetases) - catalyze condensation reactions where smaller molecules are connected with the resulting removal of a water molecule. This is accompanied by the formation of a high energy Phosphate link that stores energy. An example would be the amino acid RNA ligases.
  

SOME INDIVIDUAL ENZYMES PRODUCED BY BACTERIA IN ALKEN-MURRAY FORMULAS:

• 3,4-Dihyroxy-3,4-dihydrophenanthrene dehydrogenase - breaks 3,4-Dihyroxy-3,4-dihydrophenanthrene into 3,4-Dihydroxyphenanthrene
• Amylase - digests starch (polymers of glucose) and carbohydrates from potatoes and grains
• Acid phosphatase (a hydrolase type enzyme that catalyses the conversion of an ortho-phosphoric monoester and water to an alcohol and ortho-phosphate. Enzyme more commonly produced by gram-negative species, but recently discovered in strains of Bacillus that also produce high levels of esterase and esterase-lipase enzymes.
• Alcohol dehydrogenase (S & R) - reduce ketones.
• Alcohol oxidase - uses primary alcohol and molecular oxygen as an electron acceptor
• Aldehyde hydrogenase - together with others, breaks aliphatic hydrocarbons into fatty acids
• Alkyl sulfatase - a detergent splitting enzyme
• Alkaline phosphatase - an enzyme that scavenges phosphate from organic sources, when inorganic phosphate is limited in wastewater.
• Ammonia monoogygenase - used by Nitrosomonas europaea to convert ammonia to nitrite.
• Alpha galactosidase - digests poly and oligo-saccharides from plant origin, such as from legumes, usually can also digest sugars including raffinose, stachiose and mellibiose. Bacteria that produce this enzyme are especially useful in green-waste composting , etc. - see CF 1008, 7014, 7016, 7114 7119 & 8000..
• Beta-1, 4-Galactosyltransferase - synthesis of disaccharides.
• Beta-galactosidase - hydrolyzes ONPG to o-nitrophenol and galactose
• Beta-glucanase (aka 1,3-beta-D-glucan-3-glucanohydralase or 1,4-beta-D-glucan-4-glucanohydralase) one of three enyzmes responsible for cellulose degradation, specifically breaks down beta-linked glucose polymers often associated with grains, such as barley, oats, wheat, soy bean meal, locust bean gum, etc. Broadly classified as a "gumase" activity.
• Casease - a protease that breaks down casein in milk and cream
• Carbonyl reductase - reduces carbonyl to alcohols
• Cellulase - Degrades the main structural components of cell wall material. Degrades cellulose, thus initiating the digestion of sawdust, grass clippings, paper, toilet paper, tissue, etc. Works together with hemicellulases (xylanases) to fully digest the above-named list.
• Chymotrypsin -secondary enzyme to trypsin, usually found together from animal or bacterial source.
• Citrase - enzyme converts citrate to pyruvic acid, acetic acid and carbon dioxide, which raises the pH of the agar slant turning it blue, when positive.
• Cytochrome C. oxidase - used by Nitrobacter winogradskyl to recover energy when converting nitrite to nitrate.
• Cyanide dihydratase one of the enzymes used to convert cyanide to ammonia and carbon dioxide. Other enzymes needed to complete the pathway are: :cyanide dihydratase, Cyanide hydratase, Beta-cyanoalanine synthase & asparagene synthase.
• Esterase - breaks down "esters of short-chain fatty acids and glycerol". The most common substrates used to trigger production of the esterase enzyme are para-nitrophenyl butyrate and para-nitrophenyl acetate. bioMerieux' API-ZYM enzyme screening assay card includes test wells that test for the presence of the esterase, esterase-lipase and lipase enzymes.
• Esterase-Lipase - This enzyme is considered to be mid-way between the esterase and lipase enzymes When Tween 80, the oleic monoester of polyoxyethylene sorbitan is used as an optimal substrate to clearly demonstrate this enzyme, especially when coupled with Victoria Blue B dye that stands overnight to increase color-depth prior to pouring and inoculating agar plates. The bioMerieux API-ZYM includes a test for esterase-lipase, in addition to individual screening ampules for esterase and lipase alone.
• Hemicellulase - a class of enzymes that breaks down the plant-derived heteropolysaccharides, also identified as "xylans", which bind cellulose and lignin molecules together. The most well-known hemicellulase is xylanase, one of the enzymes included in screening tests performed by Alken-Murray Corporation.
• Hydroxylamine oxidoreductase - another enzyme used by Nitrosomonas europaea
• Hyponitrate reductase - catalyst to turn hyponitrate to nitrogen gas, part of denitrification.
• Invertase (aka Beta-fructofuranosidase) - hydrolyzes saccharose (sucrose) into glucose + fructose. Invertase is the actual enzyme that changes the pink bioMerieux API-50CHB media in the saccharose ampule to a yellow color, when the API-50CH card is read at 24 or 48 hours, post inoculation.
• Lactase - digests lactic acid in milk products. Given as supplement to people lacking this enzyme if they wish to ingest dairy foods.
• Leucine arylamidase - enzyme that breaks down amino acid leucine into acetyl acetate and acetyl-coA, More common in gram-negative bacteria, but produced by a nice selection of Alken-Murray's Bacillus strains.
• Lipase - an enzyme that digests animal and vegetable fats and oils. Lipase substrates are specifically identified as "emulsified long-chain fatty acids", which include Tributyrin, Triolein and olive oil. Since this enzyme separates fatty acids from their glycerol backbone, releasing glycerol acts as a reward to encourage greater production of lipase enzymes by cultures that enjoy eating glycerol and are capable of producing lipase to gain it. This enzyme does NOT degrade petrochemical oils.
• Lipolase - lipolytic enzyme (a particular lipase) produced for incorporation into automatic laundry detergents. Lipolase catalyses the hydrolysis of triglycerides into more soluble materials, usually a mixture of mono- and di- glycerides, glycerol and free fatty acids. Lipolase has broad activity and promotes the hydrolysis of a wide variety of fatty substances. A dry enzyme, stable for a least one year refrigerated.
• Mono-methanoxigenase - together with others, breaks aliphatic hydrocarbons into fatty acids
• Nitrate reductase - used bycertain Paracoccus pantotrophus. Nitrosomonas europaea, Bacillus pumilus, Bacillus megaterium and Bacillus licheniformis, among other strains, under anoxic or anaerobic conditions to convert nitrate to nitrite. This enzyme performs the first step in denitrification, and more strains can perform this step than can reduce nitrate all the way to nitrogen gas or nitrous oxide.
• Nitrite oxidoreductase - an enzyme of Nitrobacter winogradskyl, etc. used to convert nitrite to nitrate
• Nitrite reductase - used by a number of denitrifying bacterial strains to convert nitrite to nitrogen gas, completing denitrification. Most strains of denitrifying bacteria, including Pseudomonas, can only denitrify under anoxic conditions, preferring dissolved oxygen when it is available, however, a few rare strains of bacteria appear to prefer nitrate and nitrite over dissolved oxygen, as long as dissolved oxygen levels are moderately low. These species include Paracoccus pantotrophus and some Bacillus, which are facultative denitrifiers, capable of utilizing oxygen when it is available to them, Thiobacillus denitrificans, which is an obligate chemotrophic denitrifier, killed by high levels of dissolved oxygen. Alken-Murray also has two strains of Bacillus pumilus, both capable of utilizing oxygen, but preferring to reduce nitrate and nitrite, even when dissolved oxygen levels approach saturation.
• Nitrogenase - reduces acetylene to ethylene
• Pectinase - digests pectin in fruit, food supplement from Aspergillus niger
• Pentosanase - digests pentosan gum from wheat flour
• Peptidase - assists protease in the digestion of proteins.
• Phenanthrene dioxygenase - breaks phenanthrene into cis-3,4-Dihyroxy-3,4-dihydrophenanthrene
• Protease - digests proteins from gelatin, meat, grains & vegetable extracts, releasing amino acids and small peptides. Bacillus subtilis protease enzyme extract is approved as GRAS food supplement. Protease from several strains of Bacillus amyloliquefaciens in the Alken-Murray collection are more powerful than those produced by strains of B.subtilis.
• Pyruvate Decarboxylase - breaks down pyruvate.
• Subtilisin aka Alcalase - is a proteolytic enzyme (one subset of protease) designed to hydrolyze all kinds of proteins including hemoglobin. Subtilisin is readily soluble in water at all reasonable use concentrations. Subtilisin is active in the hydrolysis of a wide variety of proteins.. Subtilisin is also known as biosurfactant with antibiotic properties. The original name of this enzyme was taken from its source bacteria, Bacillus subtilis. Alken-Murray utilizes four (4) unrelated strains of Bacillus subtilis in various Alken-Murray products.
• Sulfide oxidase - oxidizes sulfide into sulfate or sulfuric acid
• Tannase - hydrolyses polymeric gallate into gallate and alcohol or glucose; also hydrolyses ester links in other tannins (useful for waste handling for leather manufacturing industry)
• Trypsin - peptide hydrolase, another pathway to protein digestion. Comparable to pancreatic enzyme that allows humans and animals to digest legumes and related plants.
• Urease - breaks down urea into ammonia Used in wine production,
• Xylanase - breaks down a gummy substance of the pentosan class, present in woody tissue, and yielding xylose on hydrolysis (aka wood gum). Several AMH series Bacillus, isolated from an ~ 4,000 year old humified peat soil deposit, and two AMP series strains of Bacillus amyloliquefaciens, isolated from pine forest soil in Texas, produce both xylanase and xylosidase enzymes. Xylanase is the most well-known hemicelluase enzyme. Sigma-Adrich sells Endo-Beta, 1,4- Xylanase enzyme, isolated from a Trichoderma viride fungi.

Updated 01-31-2007