Nutritional Requirements

Microorganisms obtain energy for support of biosynthesis and growth from their environment in a variety of ways nutritional requirements are complex and varied Not only are the types of microorganisms diverse (bacteria, molds, yeast), but the species and strains become very specific; the products All microorgnism requires water, source of energy, C, N, mineral elements, possibly vitamins plus oxygen if aerobic

Produce max yield of prod. or biomass per g substrate used Produce max concentration of prod. permit max rate of prod formation Permit min yield of undesired prod Consistent quality & available throughout year Cause min problems during preparation Cause min problems during production & down stream process

availability of nutrients required by microorganisms: energy, grow, production fermentation conditions: substrate pH, MC, incubation temperature stage of growth of micro-organisms presence of other competing micro-organisms Preparation & downstream process

Natural Media:

Formulated-MEDIA

Cheaper High variation/batch Unpredictable product/biomass More complex in down stream process Difficult to detect small yield improvement

Easily reproduce Specific to the strain More expensive More predictable result More simple process cheaper Easier to detect process improvement

Usually, complex media for production of new secondary metabolites are empirically formulated, since (in most cases) little is known about the microbe or biosynthesis of the desired compounds.

Essential

stage in manufacturing process Composition satisfy for supporting of cell biomass & metabolites production Adequate supply of energy for biosynthesis & cell maintenance Considering stoichiometry for growth & product formation For aerobic fermentation: C(energy sources)+N source+O2+others biomass+P+CO2+H2O+heat Product capacity min. biomass nutrition requirement Some components need for product formation but not for biomass formation Commercially, trade secret

Require materials for energy, biosynthesis of cellular matter, products in cell operation, maintenance and reproduction predominant atomic constituents of organisms, C, H, N, 0, P, and S, making up the molecules of living matter. All living cells on earth contain water as their predominant constituent. Cell: lipids, polysaccharides, nucleic acids, proteins and few common salts 40-50% C, 30-50% O2, 6-8% H2, 3-12 N

comp. require in fermentation Use for solvent within the cell Different in percentage it has some unusual properties ionizes into acid and base, and has a propensity for hydrogen bonding May need treatments Consider: pH, salts, mineral, effluent contaminant
Major

Obtain energy from their environment for support of biosynthesis and growth; either medium oxidation or light Grouping microorganism base on energy source: autotrophic & heterotropic

AUTOTROPHIC: use CO2 as principal/carbonates/simples inorganic compounds (e.g. ammonium sulfate, magnesium sulfate and sodium chloride) as C source for synthesize all their complex chemical structures (proteins, fats, carbohy-drates, vitamins, cell walls, nucleic acids, etc.) Subdivided on the basis of their ability to utilize the energy for cell growth:light (phototrophs) or the energy of chemical reactions (chemotrophs)

use organic compounds as a source of energy and organic material for synthesis of cellular components. 1. photoautotropic: light as energy source, CO2 as C source (higher plants) 2. Photoheterotrophic: light as energy source, organic comp. as C source 3. Chemoautotropic: chemical as energy source, CO2 as C source; have ability to use reduced inorganic comp as oxidizable energy sources (NH3, NO2, H2S etc) 4. Chemoheterotrophic: chemical as energy source, organic comp. as C source (fungi, great number of bacteria)

Biomass 50% C (db); skeleton of all cellular molecules reduced carbon compounds are utilized as carbon sources for building cell mass and forming product, as well as acting as an energy source Energy sources; secreted as CO2 & organic compounds Single comp for energy & C sources; add nutrients Energy used for methabolism-biosynthetic reaction & cell maintaining

Some of the energy generated during carbon metabolism is also given off as heat

Main product of fermentation & purity determine the choice of C source Carbohydrates: C, O, H & energy 0.2-25% of media 3 classes: mono-, di-, & polysacharides Breakdown of poly- & di- to simpler sugars with enzyme help major source of energy-rich comp Lab media glucose Industrial media starch, maltose, sucrose, fructose, xylose, lactose Oils & Fats Vegetable oil: fatty acid, oleic, linoleic &linolenic acid Typical oil contain~2.4x energy of glucose/weight Less volume space smaller fermenter Hydrocaarbon & their derivatives N-alkanes for organic acid, amino acid, vitamin etc energy ~ 2x C, 3x sugar

N serve to build structural proteins, functional enzymes & nucleic acids N presents in cell as amino groups As organic or inorganic sources Organic acid faster growth & fulfill microorganism requirement N supplied as ammonia salts, amonia gas, amino acids, and urea etc utilizable ammonia from the degradation of organic nitrogenous compounds such as pro-eins, peptides, or amino acids

key nutrient for aerobic microbes commonly found as a constituent of cellular water and organic compounds. Carbohydrates are excellent sources of microorganisms that are dependent on respiration for generating energy require molecular oxygen as the final hydrogen or electron low solubility of oxygen in water Oxygen transfer achieved by shaking, by sparging and agitation.

In mM level : N, P, S, K and Mg In mM level : Fe, Na, Ca, Cu, Co, Zn, Mn etc. Supplied during cell cultivation Various functions: serve in coenzyme, catalyse reactions, vitamin synthesis, cell transport very low levels; sometimes supplied from quantities occurring in water or impurities of other ingredients Primary metabolite production is usually not very sensitive to trace element concentration, but not for secondary metabolite production

metallic elements can be supplied as nutrients in the form of the cations of inorganic salts K, Mg, Ca and Fe are normally required in relatively large amounts and should normally always be included as salts in culture media Not possible to generalize ionic requirements Oxygen is always provided in water Aerobic organisms require molecular oxygen as terminal oxidizing agents to fulfill their energetic needs through aerobic respiration. For obligate anaerobes oxygen is a toxic substance

organic comp, require in very low concentration, perform specific catalytic/structural roles vitamin, sterol, amino acids, fatty acids etc Specific for certain microorganism Required to stimulates microbial growth Many natural C & N contain some growth factors Vitamins are growth factors which fulfill specific catalytic needs in biosynthesis Vitamin function as (part of ) coenzymes to catalyze many reactions The vitamins most frequently required are thiamin and biotin. Required in the greatest amounts are usually niacin, pantothenate, riboflavin, and some folic derivatives, biotin, vitamin B, and lipoic acid are required in smaller amounts