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Ultrafiltration of fermented broths and solvent extraction of antibiotics

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Abstract

Experiments of ultrafiltration with diafiltration were performed on fermented broths of benzylpenicillin and clavulanic acid produced at pilot scale at CIPAN, SA to achieve solid/liquid separation, antibiotic recovery and soluble protein retention. This study was done using a laboratorial/pilot scale ultrafiltration facility from Paterson Candy International Ltd. whose modules have a tubular configuration. PVDF membranes with a molecular weight cut-off of 100 000 (FP10) and 200 000 (FP200) were assessed. Comparison between solvent extraction of benzylpenicillin and clavulanic acid from filtered broth and ultrafiltered broth was also done. It was concluded that ultrafiltration with diafiltration of fermented broths of antibiotics using the FP10 membrane is a valid alternative to the use of rotary vacuum filters, flocculants and filter aid, deserving a further study to establish the best operating conditions. An economic appraisal also seems to be of paramount importance.

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Abbreviations

CF:

volumetric concentration factor

C mg/1:

concentration

¯J 1/h m2 :

average permeate flux

J(t) 1/h m2 :

permeate flux at time t

Mass WB g:

mass of fermented broth

P i :

bar inlet pressure of feed

R BP %:

benzylpenicillin recovered by mass balance to the permeate, defined by equation (4)

R CA%:

clavulanic acid recovered by mass balance to the permeate, defined by equation (4)

ℜsolprot%:

retained soluble protein by mass balance to the permeate, defined by equation (5)

SI%:

solvent increase

SL%:

solvent losses

T °C:

temperature

t min:

time

Δt :

min time difference

Titre IU/g or IU/ml:

concentration of benzylpenicillin

Titre μg/ml:

concentration of clavulanic acid

V 1 or ml:

volume

η%:

yield of antibiotic

μ cp:

fluid viscosity

BP:

benzylpenicillin

CA:

clavulanic acid

FP10:

100 000 MWCO ultrafiltration membrane

FP200:

200 000 MWCO ultrafiltration membrane

FB:

filtered broth

IU:

international unit of activity

ML:

mother liquors, i.e., extracted liquors

MWCO:

molecular weight cut-off

PCI:

Paterson Candy International Ltd.

Prot FB:

protein in the filtered broth

PVDF:

polyvinylidene fluoride

RS:

rich solvent

RFB:

rich filtered broth

UF:

ultrafiltration

1 V diaf :

one diafiltration volume

WB:

fermented broth (whole broth)

C :

concentrate

FB:

filtered broth

Prot:

protein

Prot FB:

protein in the filtered broth

0:

initial value

P :

permeate

Ext:

extraction

Fil:

filtration

UF:

ultrafiltration

ML:

mother liquors, i.e., extracted liquors

G :

global

RS:

rich solvent

WB:

fermented broth (whole broth)

References

  1. Hersbach, G.J.M.; ZZZVan der Beek, C.P.; ZZZVan Dijck, P.W.M.: The Penicillins: properties, biosynthesis and fermentation. In: Vandamme, E.J. (Ed.): Biotechnology of industrial antibiotics, vol. 22, pp 45–119. New York: Marcel Dekker, Inc. 1984

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  2. Butterworth, D.: Clavulanic acid: properties, biosynthesis and Fermentation. In: Vandamme, E.J. (Ed.): Biotechnology of industrial antibiotics, vol. 22, pp 225–234. New York: Marcel Dekker, Inc. 1984

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  3. Paterson Candy International, Ltd.: Cleaning, sterilization and preservation procedures, S05 Cleaning 041 (1988)

  4. Code of Federal Regulations, CFR. 21§ 436.205

  5. Bird, A.E.; Bellis, J.M.; Gasson, B.C.: Spectrophotometric assay of Clavulanic acid by reaction with imidazole. Analyst. 107 (1982) 1241–1245

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  6. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye. Anal. Biochem. 72 (1976) 248–254

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Author information

Authors and Affiliations

  1. Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, 1000, Lisboa, Portugal

    A. M. A. Nabais & J. P. Cardoso

  2. Companhia Industrial Produtora de Antibióticos, CIPAN, SA, Vala do Carregado, 2580, Carregado, Portugal

    J. P. Cardoso

Authors
  1. A. M. A. Nabais
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  2. J. P. Cardoso
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Additional information

To CIPAN-SA in which this work was done and where the fermented broths were produced. To JNICT/FSE for the funding of the project in the scope of which this work was done.

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Nabais, A.M.A., Cardoso, J.P. Ultrafiltration of fermented broths and solvent extraction of antibiotics. Bioprocess Engineering 13, 215–221 (1995). https://doi.org/10.1007/BF00367257

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  • DOI: https://doi.org/10.1007/BF00367257

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