Bacteria protein production

Streptokinase. The fibrinolytic activity of streptokinase, isolated from strains of hemolytic Streptococci, was first demonstrated in 1933 (63). Streptokinase is a secreted protein product inasmuch as filtrates free of demonstrable bacteria were found to dissolve fibrin clots with rapidity. 

Other Gram-positive food-grade bacteria, such as the corynebacterium Corynebacterium glutamicum, may soon become attractive hosts for heterologous protein production due to low... 

At the moment, most of these more exotic bacteria are mainly used in expression for fundamental studies. Considering the speed of developments in biotechnology, their application for industrial or pharmaceutical protein production might become even more important very soon. 

Host Cell Impurities Various organisms have been used to produce recombinant proteins yeast, bacteria (e.g., E. coli), insect cells, and mammalian cells such as Chinese hamster ovary (CHO) cells. During the purification process, some HCPs can copurify with the protein product. Because of the specificity of the antigen-antibody interaction, an ELISA can be used to detect and quantitate the contaminating HCPs. Detecting host impurities is important for quality process control as well as for product safety issues. The intent is to avoid unsafe levels of residual HCPs which might lead to adverse reactions.11... 

J. Lee and W.F. Ramirez. Mathematical modeling of induced foreing protein production by recombinant bacteria. Biotechnol. Bioeng., 39 635-, 1992. 

Of the particulate stimuli certain ones are far more active when they are coated with proteins from serum (opsonized) than when they are not. However, others like latex beads elicit formation of Oj" by PMNs without opsonization DeChatelet et al. found that the production of O by PMNs from man and rabbit was stimulated by opsonized but not unopsonized zymosan (fragments of yeast cell walls). Bacteria alone were found to stimulate the formation of O but in the presence of serum bacteria stimulated the formation of O7 three fold However, the stimulatory effect of bacteria appeared to be caused by changes which the bacteria produced by an interaction with constitutents of serum, because serum itself after exposure to the bacteria stimulated production of O by PMNs. The active component from serum was heat sensitive (100°) and not sedimentable at 105,000 g. Whether this material was derived from the components of serum or from the bacteria is not clear but may have been a protein of the complement system. 

Discuss the changes that must be made in a typical eukaryotic structural gene to allow its protein product to be synthesized in bacteria. 

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