Beruflich Dokumente
Kultur Dokumente
5]
(a) Substrate
appeared to be the most satisfactory other nutrients are in excess. As the except that both glucose and prop-
candidate. It was easy to grow, culture grows, the phosphate supply ionic acid are fed in the second
accumulated high levels of high becomes depleted and the culture stage. The hydroxyvaleric content of
molecular weight polymer (up to enters phosphate limitation; little the polymer is controlled by varia-
80% of cell dry weight) 19 and used a PHB has been accumulated in the tion of the ratio of glucose to
range of substrates, some of which cells up to this point in the fermenta- propionate in the feed. Good control
were potentially very attractive for tion. During the next 48 hours, of propionate feed rate is essential,
PHB production. glucose is added to the culture and since a supernatant concentration in
At first sight, the use of hydrogen/ the dry weight of the biomass rises excess of 0.1% is toxic and prevents
carbon dioxide/air as a fermentation considerably due to massive PHB polymer synthesis.
substrate looks an economically accumulation by the cells. Concen- The copolymers have superior
viable alternative. An investiga- trations of PHB up to 75% of the total mechanical properties to the homo-
tion of the engineering problems dry weight of the culture can be polymer. They are more flexible and
involved in building plant and oper- achieved. tougher which extends their versati-
ating a fermentation with flammable The initial growth phase of the lity in end use to applications such
hydrogen/air mixtures, however, fermentation to the point where as bottle and film manufacture
forces the conclusion that the high phosphate limitation is reached which are not possible with PHB.
capital cost of such an installation takes approximately 60 hours. The
would make the process unecono- total fermentation time, including Product recovery
mic. the glucose feeding phase, is in the Polymer can be removed from the
Ethanol gives good yields but is order of 110 to 120 hours. biomass in a number of ways.
more expensive than carbohydrates, -Fig. 3 Through solvent extraction, recover-
as is acetic acid. Therefore, sucrose Cells ies of greater than 90% of the
as either cane sugar or molasses is from fermenter polymer in cells can be achieved.
The cells are refluxed in hot meth-
apparently the best carbon and
energy source.
Additional processing of these
I anol to remove lipids and phospho-
lipids and then extracted with chloro-
Cell ]
carbohydrate sources would be rupture form or methylene chloride. PHB is
necessary: in both cases the sucrose soluble in these solvents. The solu-
would have to be 'inverted' (con-
verted into glucose and fructose
I tion is filtered hot to remove cell
debris and cooled to precipitate
sugar monomers) and molasses
might have to be purified to reduce
I 'nz'°e
treatment I PHB. The PHB, a low bulk density
white powder, is finally vacuum
the nitrogen and phosphorus con-
centration so that fermentations I dried 2°. The process is expensive,
requires large inventories of solvent
could be run under nitrogen- or
phosphorus-limited conditions. Dex- [ Was.ing ] and heavy capital investment in
solvent recovery plant.
trose would also be an attractive Development of an alternative
substrate - it can be obtained in a I non-solvent-based extraction pro-
pure form, in large quantities and Drying cess has been necessary to make the
requires no additional processing. Extrusion overall PHB production process
Unfortunately for those companies Comminution more economically attractive. Cells
operating in the EEC, the Common are ruptured and the resultant debris
Agricultural Policy has meant that I is treated with an enzyme cocktail to
carbohydrate substrates are more Polymer chips solubilize the non-PHB components.
expensive for the EEC chemical Recovery and separation of poly-
After washing and flocculation, PHB
industry than for their competitors mer (PHB) from cell fermentation is recovered as white powder. The
who can obtain sugar at world prices process and subsequent conver- powder is converted to polymer
(Table 3). Nevertheless, growing sion to polymer chips. chips in a final blending, melt
Alcaligenes eutrophus on glucose extrusion, comminution process
still seems the best alternative for a Two fermenter types have been (Fig. 3).
production process. successfully used for PHB produc-
tion: a 35 000 litre air-lift fermenter The future
Production technology and various sizes of conventional The number of microbially pro-
ABcaligenes eutrophus is cur- stirred vessels up to 200 000 litres. duced biopolymers exploited by
rently grown in a glucose-salts industry is small. They are, in the
medium in a fed batch reactor. The Copolymer synthesis main, the obvious products excreted
phosphate content of the medium is If a copolymer of PHB and PHV is or accumulated by a range of rela-
such that it will support only a required, the fermentation is essen- tively well-known microorganisms.
certain amount of cell growth: all tially similar to that described above It seems likely that there are other
TIBTECH - SEPTEMBER 1987 [Vol. 5]
materials waiting to be discovered in 4 Kennedy, J.F. and Bradshaw, L.J. 13 De Smit M., Eggink, G., Witholt, B.,
nature - one has only to draw the (1984) in Progress in Industrial Micro- Kingman, J. and Wynberg, H. (1983)
analogy with the vast range of biology (Vol. 19), (Bushell, M.E., J. Bacteriol. 154, 870-878
antibiotics w h i c h were discovered ed.), pp. 319-371, Elsevier 14 Ajsenjo, J. A. and Snk, J. S. (1986) J.
once the first few h a d been 5 0 g a t a , K. (1975) in Advances in Ferment. Technol. 64, 271-278
characterized. Modern genetic tech- Applied Microbiology (Vol. 19) (Perl-
man, D., ed.), pp. 209-247, Academic 15 Oeding, V. and Schlegel, H. G. (1973)
niques enable copies or analogues of Biochem. J. 134, 239-248
Press
some natural materials to be synthes- 6 Holmes, P. A. (1985) Phys. Techno]. 16 Collins, S. H. (1987) in Carbon Sub-
ized and the biosynthetic p a t h w a y to 16, 32-36 strates in Biotechno]ogy (Vol. 21),
others to be m a n i p u l a t e d to advan- 7 Genetic.. Technology News (1987) (Stowe]l, J. D., Beardsmore, A. J.,
tage. The future for biopolymers (March), 6-7 Keevil, C. W. and Woodward, J. R.,
looks exciting provided that t h e y can 8 Dawes, E. A. and Senior, P. J. (1973) eds), pp. 161-169, IRL Press
fulfil requirements of price and Adv. Microb. Physiol. 10, 135-266 17 Susuki, T., Yamano, T. and Shimigu,
performance. 9 Martin, A., Veldhuis, C., Stegeman, S. (1986) Microbio]. Biotechno]. 23,
V. and Veenhuis, S. (1979) ]. Gen. 322-329
Microbio]. 112,249-355
References 10 Holmes, P.A., Wright, L.E. and 18. Holmes, P. A. in Developments in
1 Kang, K. S. and Cottrell, I. W. (1979) Crystalline Polymers (Vol. 2) (Basset,
Collins, S. H. (1981) European Patent
in Microbial Technology (2nd edn) 0 052 459 D. C., ed.), Applied Science Pub-
(Peppier, H. J. and Perlman, D. eds), lishers (in press)
11 Findlay, R.H. and White, D.C.
Academic Press (1983) App]. Environ. Microbio]. 45, 19 Heinzle, E. and Lafferty, R. M. (1986)
2 Jarman, T. R. (1982) European Patent 71-78 Eur. J. Appl. Microbio]. Biotechno].
0 066 377 12 Wallen, L. L. and Rohwedder, W. K. 11, 8-16
3 Bracke, J. W. and Thacker, K. (1985) (1974) Environ. Sci. Technol. 8, 576- 20 Stageman, J.F. (1983) European
US Patent 4 517 295 579 Patent Application 124 309
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