Microbial cell

1 It is recommended that readers obtain any college-level textbooks in biology and microbiology for further reading and reference as the need occurs. 

Unicellular Protists Algae Protozoa Fungi Molds Yeasts  

Cell wall Cell membrane Ribosome Nuclear region Cytoplasm 

The prokaryotic cell is surrounded with a cell wall and a cell membrane. The cell wall, considerably thicker than the cell membrane, protects the cell from external influences. The cell membrane (or cytoplasmic membrane) is a selective barrier between the interior of the cell and the external environment. The largest molecules known to cross this membrane are DNA fragments and low-molecular-weight proteins. The cell membrane can be folded and extended into the cytoplasm or internal membranes. The cell membrane serves as the surface onto which other cell substances attach and upon which many important cell functions take place. 

Clostridium, a small spindle Pasteurella after Louis Pasteur, Latinized Salmonella after Daniel E. Salmon, Latinized Saccharomyces sugar fungus 

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Modem technology for produciag microbial cells for human food or animal feed emerged ia Germany duriag World War I. Baker s yeast,... 

Experimental procedures have been described in which the desired reactions have been carried out either by whole microbial cells or by enzymes (1—3). These involve carbohydrates (qv) (4,5) steroids (qv), sterols, and bile acids (6—11) nonsteroid cycHc compounds (12) ahcycHc and alkane hydroxylations (13—16) alkaloids (7,17,18) various pharmaceuticals (qv) (19—21), including antibiotics (19—24) and miscellaneous natural products (25—27). Reviews of the microbial oxidation of aUphatic and aromatic hydrocarbons (qv) (28), monoterpenes (29,30), pesticides (qv) (31,32), lignin (qv) (33,34), flavors and fragrances (35), and other organic molecules (8,12,36,37) have been pubflshed (see Enzyp applications, industrial Enzyt s in organic synthesis Elavors AND spices). 

Enzymatic Process. Chemically synthesized substrates can be converted to the corresponding amino acids by the catalytic action of an enzyme or the microbial cells as an enzyme source, t - Alanine production from L-aspartic acid, L-aspartic acid production from fumaric acid, L-cysteine production from DL-2-aminothiazoline-4-catboxyhc acid, D-phenylglycine (and D-/> -hydtoxyphenylglycine) production from DL-phenyUiydantoin (and DL-/)-hydroxyphenylhydantoin), and L-tryptophan production from indole and DL-serine have been in operation as commercial processes. Some of the other processes shown in Table 10 are at a technical level high enough to be useful for commercial production (24). Representative chemical reactions used ia the enzymatic process are shown ia Figure 6. 

Sodium chlorite oxidation of com and rice starches is recommended for the production of textile sizes (101) and oxidized starch is recommended as a hardening agent in the immobilization of microbial cells within gelatin (102). 

It is necessary to estabUsh a criterion for microbial death when considering a sterilization process. With respect to the individual cell, the irreversible cessation of all vital functions such as growth, reproduction, and in the case of vimses, inabiUty to attach and infect, is a most suitable criterion. On a practical level, it is necessary to estabUsh test criteria that permit a conclusion without having to observe individual microbial cells. The failure to reproduce in a suitable medium after incubation at optimum conditions for some acceptable time period is traditionally accepted as satisfactory proof of microbial death and, consequentiy, stetihty. The appHcation of such a testing method is, for practical purposes, however, not considered possible. The cultured article caimot be retrieved for subsequent use and the size of many items totally precludes practical culturing techniques. In order to design acceptable test procedures, the kinetics and thermodynamics of the sterilization process must be understood. 

N. S. Ryder ia C. Nombela, ed.. Microbial Cell Wall Synthesis andMutolysis Elsevier Science PubHshers, Amsterdam, The Netherlands, 1984, pp. 

Recombinant DNA technology has already provided several products of therapeutic interest from mammalian cells. Table 2 gives examples of products from mammalian cells, the use, and the technology used for production. Technology development for these products has centered around the differences in characteristics of mammalian versus microbial cells, notably, the shear sensitivity and susceptibiUty to contamination of the mammalian lines. 

Considerable work has been done to try to explain why quats are antimicrobial. The following sequence of steps is beheved to occur in the attack by the quat on the microbial cell (/) adsorption of the compound on the bacterial cell surface (2) diffusion through the cell wall (J) binding to the cytoplasmic membrane (4) dismption of the cytoplasmic membrane (5) release of cations and other cytoplasmic cell constituents (6) precipitation of cell contents and death of the cell. 

Recovery. The principal purpose of recovery is to remove nonproteinaceous material from the enzyme preparation. Enzyme yields vary, sometimes exceeding 75%. Most industrial enzymes are secreted by a microorganism, and the first recovery step is often the removal of whole cells and other particulate matter (19) by centrifugation (20) or filtration (21). In the case of ceU-bound enzymes, the harvested cells can be used as is or dismpted by physical (eg, bead mills, high pressure homogenizer) and/or chemical (eg, solvent, detergent, lysozyme [9001 -63-2] or other lytic enzyme) techniques (22). Enzymes can be extracted from dismpted microbial cells, and ground animal (trypsin) or plant (papain) material by dilute salt solutions or aqueous two-phase systems (23). 

Because enzymes can be intraceUularly associated with cell membranes, whole microbial cells, viable or nonviable, can be used to exploit the activity of one or more types of enzyme and cofactor regeneration, eg, alcohol production from sugar with yeast cells. Viable cells may be further stabilized by entrapment in aqueous gel beads or attached to the surface of spherical particles. Otherwise cells are usually homogenized and cross-linked with glutaraldehyde [111-30-8] to form an insoluble yet penetrable matrix. This is the method upon which the principal industrial appHcations of immobilized enzymes is based. 

FIG. 18-28 Usually, the gas-liquid mass-transfer coefficient, K, is reduced with increased viscosity. This shows the effect of increased concentration of microbial cells in a fermentation process. 

Plant and animal cells have numerous chromosomes. Growth rates are relatively slow. A typical nutrient medium will contain a large number of vitamins and growth factors in addition to complex nitrogen sources, because other specialized cells in the original structures supply these needs. A plant or animal cell is not hke a microbial cell in its ability to function independently. 

Mammalian Cells Unlike microbial cells, mammalian cells do not continue to reproduce forever. Cancerous cells have lost this natural timing that leads to death after a few dozen generations and continue to multiply indefinitely. Hybridoma cells from the fusion of two mammalian lymphoid cells, one cancerous and the other normal, are important for mammalian cell culture. They produce monoclonal antibodies for research, for affinity methods for biological separations, and for analyses used in the diagnosis and treatment of some diseases. However, the frequency of fusion is low. If the unfused cells are not killed, the myelomas 1 overgrow the hybrid cells. The myelomas can be isolated when there is a defect in their production of enzymes involved in nucleotide synthesis. Mammahan cells can produce the necessary enzymes and thus so can the fused cells. When the cells are placed in a medium in which the enzymes are necessaiy for survival, the myelomas will not survive. The unfused normal cells will die because of their limited life span. Thus, after a period of time, the hybridomas will be the only cells left ahve. 

Monod kinetics Kinetics of microbial cell growth as a function of substrate concentration proposed by Jacques Monod and widely used to understand growth-substrate relationships. 

In most animal, plant, and microbial cells, the enzyme that phosphorylates glucose is hexokinase. Magnesium ion (Mg ) is required for this reaction, as for the other kinase enzymes in the glycolytic pathway. The true substrate for the hexokinase reaction is MgATP. The apparent K , for glucose of the animal... 

The resultant fermentation broth was centrifuged to harvest the microbial cells, and they were washed with water and centrifuged a second time, whereupon a living cell paste was obtained. (There was obtained an amount of cells equivalent to 54 parts on a dry basis, which contained 920 /Jg of ubiquinone-10 per gram of dry cells.)... 

Microbial cells are very attractive as a source of catalysts for the production of organic chemicals because of their broad range of enzymes capable of a wide variety of chemical reactions, some of which are illustrated in Table 2.1. 

Microbial cells, rather than plant and animal cells, are generally preferred for the production of organic chemicals. There are several reasons for this. 

We can also use microbial cells (fermentation) containing the desired catalytic activity without isolating the enzymes responsible. 

Synthesis of industrial chemicals by microbial cells may be by fermentation (free, living cells), immobilised growing cells, immobilised resting cells or immobilised dead cells. 

Several of the problems associated with whole cell bioprocesses are related to the highly effective metabolic control of microbial cells. Because cells are so well regulated, substrate or product inhibition often limits the concentration of desired product that can be achieved. This problem is often difficult to solve because of a poor understanding of the kinetic characteristics of the metabolic pathway leading to the desired product. 

Single cell protein, normally called simply SCP, is the term used to describe microbial cells, or proteins from them, which are used as food (food for humans) or feed (food for farm animals or fish). Although the term micro-organisms covers viruses, bacteria, fungi, algae and protozoa, viruses and protozoa are not considered suitable for SCP production. 

Micro-organisms are rich in protein. Microbial cells can contain as much protein as conventional foods. Bacteria can contain 60-65% (as a % of dry weight) protein whereas fungi and algae contain about 40%. In addition, microbial cells can be a rich source of fibre, unsaturated fats, minerals and vitamins. They are low in saturated fats and sodium. 

What are they like to eat Humans are particular about the organoleptic properties of their food. Microbial cells may have little taste or smell, or even smell or taste unpleasantly to some people. The texture may not be the same as in conventional foods, particularly with unicellular organisms. These draw-backs can be overcome by adding a proportion of SCP to manufactured foods. However, even when SCP is incorporated into manufactured foods it may not have suitable characteristics such as stability, ability to bind water or fats, or ability to form gels, emulsions or foams. SCP for feed does not have to meet such strict requirements. 

Study the unlabelled block diagram, and then replace the question marks with the words and phrases to give a generalised scheme of an industrial fermentation. Assume in this example that the product is excreted from the microbial cells. 

Many different types of carbohydrate-containing molecules are located on the surface of microbial cells. Some of these are components of die microbial cell wall and are limited to certain types of micro-organisms such as bacterial peptidoglycan, lipopolysaccharides, techoic adds and yeast mannans. Other polysaccharides are not... 

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