Biopolym. Cell. 2002; 18(2):164-170.
Molecular and Cell Biotechnologies
Investigation the influence of cultivation conditions on production of human alpha-2b interferon in Escherichia coli
1Slavchenko I. Yu.
  1. PSRS "Biotechnolog"
    150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680

Abstract

The influence of glucose and maltose presence in a growth medium, as well as the temperature of producer cultivation on the production level of the human alpha-2b interferon (1FN) in E. coli have been studied. E. coli SG30 (pIF-16) harbouring plasmid pIF-16 bearing tandem of the artificial IFNs genes under the control of the trp promoters' tandem has been used as strain-producer of IFN. Results indicate that during E. coli SG30 (pIF-16) cells cultivation at 37 degrees C in the presence of maltose or glucose as carbone sourse in growth medium the yield of IFN was appreciably decreased in comparison with carbons – free medium. We have also demonstrated that the level of IFN production depends on the concentration of these carbons in a medium and the cultivation temperature of producer. Glucose exerts negative influence on synthesis IFN In lower concentration, than maltose. It was shown this study, that the addition of maltose to the medium resulted higher yield of cells biomass, than the addition of glucose during growth E. coli SG30 (pIF-16) both 37, and 28 degrees C.

References

[1] Mashko SV. Optimization expression of foreign genes in E. coli cells. Biotekhnologiya. 1998. 6:3-23.
[2] Baneyx F. Recombinant protein expression in Escherichia coli. Curr Opin Biotechnol. 1999;10(5):411-21.
[3] Weickert MJ, Doherty DH, Best EA, Olins PO. Optimization of heterologous protein production in Escherichia coli. Curr Opin Biotechnol. 1996;7(5):494-9.
[4] Jacques N, Guillerez J, Dreyfus M. Culture conditions differentially affect the translation of individual Escherichia coli mRNAs. J Mol Biol. 1992;226(3):597-608.
[5] Murby M, Uhl?n M, St?hl S. Upstream strategies to minimize proteolytic degradation upon recombinant production in Escherichia coli. Protein Expr Purif. 1996;7(2):129-36.
[6] Winther-Larsen HC, Josefsen KD, Brautaset T, Valla S. Parameters affecting gene expression from the Pm promoter in gram-negative bacteria. Metab Eng. 2000;2(2):79-91.
[7] Heyde M, Laloi P, Portalier R. Involvement of carbon source and acetyl phosphate in the external-pH-dependent expression of porin genes in Escherichia coli. J Bacteriol. 2000;182(1):198-202.
[8] Chagneau C, Heyde M, Alonso S, Portalier R, Laloi P. External-pH-dependent expression of the maltose regulon and ompF gene in Escherichia coli is affected by the level of glycerol kinase, encoded by glpK. J Bacteriol. 2001;183(19):5675-83.
[9] Tseng CP, Yu CC, Lin HH, Chang CY, Kuo JT. Oxygen- and growth rate-dependent regulation of Escherichia coli fumarase (FumA, FumB, and FumC) activity. J Bacteriol. 2001;183(2):461-7.
[10] Tao H, Bausch C, Richmond C, Blattner FR, Conway T. Functional genomics: expression analysis of Escherichia coli growing on minimal and rich media. J Bacteriol. 1999;181(20):6425-40.
[11] Ertl P, Unterladstaetter B, Bayer K, Mikkelsen SR. Ferricyanide reduction by Escherichia coli: kinetics, mechanism, and application to the optimization of recombinant fermentations. Anal Chem. 2000;72(20):4949-56.
[12] Jeong KJ, Lee SY. High-level production of human leptin by fed-batch cultivation of recombinant Escherichia coli and its purification. Appl Environ Microbiol. 1999;65(7):3027-32.
[13] Schmidt M, Babu KR, Khanna N, Marten S, Rinas U. Temperature-induced production of recombinant human insulin in high-cell density cultures of recombinant Escherichia coli. J Biotechnol. 1999;68(1):71-83.
[14] Charpentier B, Bardey V, Robas N, Branlant C. The EIIGlc protein is involved in glucose-mediated activation of Escherichia coli gapA and gapB-pgk transcription. J Bacteriol. 1998;180(24):6476-83.
[15] Li X, Taylor KB. Effect of glucose on the expression parameters of recombinant protein in Escherichia coli during batch growth in complex medium. Biotechnol Prog. 1994;10(2):160-4.
[16] Brautaset T, Petersen S, Valla S. An experimental study on carbon flow in Escherichia coli as a function of kinetic properties and expression levels of the enzyme phosphoglucomutase. Biotechnol Bioeng. 1998 Apr 20-May 5;58(2-3):299-302.
[17] Aristidou AA, San KY, Bennett GN. Improvement of biomass yield and recombinant gene expression in Escherichia coli by using fructose as the primary carbon source. Biotechnol Prog. 1999;15(1):140-5.
[18] Kagawa N, Cao Q. Osmotic stress induced by carbohydrates enhances expression of foreign proteins in Escherichia coli. Arch Biochem Biophys. 2001;393(2):290-6.
[19] Barth S, Huhn M, Matthey B, Klimka A, Galinski EA, Engert A. Compatible-solute-supported periplasmic expression of functional recombinant proteins under stress conditions. Appl Environ Microbiol. 2000;66(4):1572-9.
[20] Blackwell JR, Horgan R. A novel strategy for production of a highly expressed recombinant protein in an active form. FEBS Lett. 1991;295(1-3):10-2.
[21] Slavchenko IYu. Study bacteriophage L efficiency for human a2 interferon production in Escherichia coli. Auth. Thesis. ... kand biol nauk. Kiev, 1990. 19 p.
[22] Slavchenko IYu. Isolation of clones sensitive to bacteriophage ? from the phage resistant Escherichia coli strain. Biopolym Cell. 2001; 17(2):160-5.
[23] Slavchenko IYu. The expression of human alpha-2b interferon in different strains of Escherichia coli. Biopolym Cell. 2001; 17(6):546-50.
[24] Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab, 1982; 545 p.
[25] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[26] AS. SSSR N 1092176. A method for preparing an artificial human interferon gene al and method for producing a polypeptide with an interferon activity microbiological synthesis. MN Kolosov, VG Korobko, VN Dobrynin, IV Severtsova, SA Chuvpilo, NS Bystrov, YuA Berlin, AL Kayushin, VV Butkus, IA Polyakova, EF Boldyreva, LS Sandakchiev, SG Popov, TN Shubina, VV Kravchenko, OI Serpinskiy, VF Yamschikov, SI Belikov, AN Sinyakov, GF Sivolobova. Publ. in BI N 18, 1984.
[27] Kravchenko VV, Guileva IP, Shamin VV, Kulichkov VA, Dobrynin VN, Filippov SA, Chuvpilo SA, Korobko VG. Construction of a tandem of artificial genes encoding human leukocyte interferon and its expression as a part of polycistronic template with coupled translation. Bioorg Khim. 1987; 13 (9):1186-93.
[28] Akimenko ZA, Zykov SA, Shaprov VV, Ofitserov VI, Gileva IP, Kravchenko VV, Sandakhchiyev LS. [A chemically synthesized gene assures the biosynthesis of a polypeptide in Escherichia coli cells, the structure of which corresponds to human leukocytic interferon alpha 2]. Dokl Akad Nauk SSSR. 1991; 319(5):1248-51.
[29] Schwartz M. The adsorption of coliphage lambda to its host: effect of variations in the surface density of receptor and in phage-receptor affinity. J Mol Biol. 1976;103(3):521-36.
[30] Kolb A, Busby S, Buc H, Garges S, Adhya S. Transcriptional regulation by cAMP and its receptor protein. Annu Rev Biochem. 1993;62:749-95.