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Disruption of microbial cells for intracellular products
YUSUF CHISTI and MURRAY MOO-YOUNG*
Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario, CanadaN2L 3GI
Summary. Disintegration of microbial cells is a necessary first step for the production of intracellular enzymes and organelIes. With increasing use of intracellular microbial material in industry and medicine, the cell disruption unit operation is gaining in importance.
This review examines the state of the art of the large-
scale cell disruption technology and disruption methods of potential commercial value.
Keywords.. Disruption of microorganisms; cell disintegration; intracellular enzymes
The importance of microorganisms as a source of commer- cially useful chemicals, antibiotics and enzymes has been recognized for a very long time. Nearly all chemicals of microbial origin produced industrially today are of the extracellular type. That is, they are produced within the microbial cell, but are then excreted into the surrounding environment. A much larger proportion of the potentially useful microbial products is retained within the cells. A vast majority of the enzymes known, for example, are intra- cellular.1 Even greater use of microbial products, many of which will be intracellular, can be expected from the predicted surge in biotechnology.2
The isolation of intracellular material requires that the
*To whom correspondence should be addressed
cell either be genetically engineered so that what would normally be an intracellular product is excreted into the environment, or it must be disintegrated by physical, chemical or enzymatic means to release its contents into the surrounding medium. The genetic manipulation of microbial cells to make them leaky is limited in scope. Making the ceil fully permeable to any significant fraction of the intracellular products and enzymes would not only be difficult, but also will imply discontinued existence of the cell. It is in this context that the unit operation of microbial cell disruption for intracellular product isolation will become of increasing importance.
Probably because of the high capital and operating costs of pilot plants for large-scale isolation of intracellular products and the requirement of sizeable teams of scientists and technical staff to obtain meaningful biochemical engineering design data, few studies have been published on the subject.3 This review examines the current state of microbial cell disruption technology from an industrial applications point of view. The disruption techniques of potential industrial use are also discussed.
In the last few years several intracellular enzymes have begun to be produced industrially: for example, glucose oxidase for food preservation, penicillin acylase for anti- biotic conversion, and asparaginase for possible cancer therapy.4 Other examples of intracellular microbial enzymes produced commercially are given in Table l.
The necessity of harvesting the producing cells, in order subsequently to extract an internal constitutent, is a major economic disadvantage and, in part, results in the present preoccupation with the manufacture of pro- ducts of very high value,s Possible cost reductions, how- ever, may be achieved by simultaneous isolation of a number of intracellular products following cell disruption.6 This would be even more so if extracelhilar product isola- tion preceded the intracellular product isolation from the same fermentation batch.
Table 1 Some examples of intracellular microbial enzymes produced commercially. [Reproduced from Lilly, M. D. in Applied Biochemistry and Bioengineering (Wingard Jr, L. 8., Katchalski.Katzir, E. and Goldstein, L., eds) Academic Press, New York, 1979, vo[. 2, p. 1 by permission of Academic Press ©]
L-AsparagJnase (EC 188.8.131.52 )
Catalase (EC 184.108.40.206) Cholesterol oxidase (EC 220.127.116.11) /3-Galactosidase (EC 3,2.1.23)
Glucose isomerase (EC 18.104.22.168) Glucose oxidase (EC 22.214.171.124)
Glucose-6-phosphate dehydrogenase (EC 126.96.36.199) Invertase (EC 188.8.131.52)
Penicillin acylase (EC 184.108.40.206 )
194 Enzyme Microb. Technol. 1986, vol. 8, April
Erw/nia carotovora Escherichia coil Aspergillus niger Nocardia rhodochrous Kluyveromyces fragilis Saccharomyces lactis Bacillus coagulans
Streptomyces sp. Aspergil/us niger
Saccharomyces cerevisiae Escherichia coil
Examples of use
Treatment of acute lymphatic leukaemia
Removal of H=O: after milk sterilization Serum cholesterol analysis
Hydrolysis of lactose in milk/whey
Production of high-fructose glucose syrups
Serum glucose analysis Removal of oxygen from foods Clinical analysis
Deacylation of benzylpenicillin
© 1986 Butterworth & Co. (Publishers) Ltd
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