EFFECT OF PREINDUCTION SPECIFIC GROWTH-RATE ON RECOMBINANT ALPHA-CONSENSUS INTERFERON SYNTHESIS IN ESCHERICHIA-COLI

被引：57

作者：

CURLESS, C ^{[1
]}

POPE, J ^{[1
]}

TSAI, L ^{[1
]}

机构：

[1] AMGEN INC,1900 OAK TERRACE LANE,THOUSAND OAKS,CA 91320

来源：

BIOTECHNOLOGY PROGRESS | 1990年 / 6卷 / 02期

关键词：

D O I：

10.1021/bp00002a009

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The effect of preinduction specific growth rate on the yield of recombinant alpha consensus interferon in Escherichia coli K‐12 was investigated. The cells used in the investigation contain a temperature‐sensitive, walkaway plasmid bearing an insert that codes for alpha consensus interferon. Transcription of the recombinant gene is controlled by a lambda repressor/pL promoter system. The lambda promoter is regulated by the temperature‐sensitive gene cl857 at 30 °C, but at 42 °C the promoter is derepressed. The cells were grown under glucose‐limited conditions in a chemostat at pH 7 and 30 °C. Once steady state was achieved, the vessel temperature was raised to 42 °C and a fed‐batch mode was initiated. Six dilution rates ranging from 0.025/h to 0.2/h were investigated. Cell dry weight, alpha consensus interferon content, glucose concentration, acetate concentration, and plasmid stability were measured. At each dilution rate, the expression level of alpha consensus interferon (g/g of cell dry wt) reached its maximum value approximately 3 h after induction. In addition, the expression level of alpha consensus interferon increases 4‐fold as the dilution rate prior to induction is increased from 0.025/h to 0.2/h. Consequently, the expression of recombinant protein produced by E. coli is dependent on the preinduction specific growth rate. Copyright © 1990 American Institute of Chemical Engineers (AIChE)

引用

页码：149 / 152

页数：4

共 11 条

[1]

Goeddel D.V., Heyneker H.L., Hozumi T., Arentzen R., Itakura K., Yansura D.G., Ross M.J., Miozzari G., Crea R., Seeburg P.H., Nature, 281, pp. 544-548, (1979)

[2]

Goeddel D.V., Yelverton E., Ulrich A., Heyneker H.L., Miozzari G., Holmes W., Seeburg P.H., Dull T., May L., Stebbing N., Crea R., Maeda S., McCandliss R., Sloma A., Tabor J.M., Gross M., Familetti P.C., Pestka S., Nature, 278, pp. 411-416, (1980)

[3]

Remaut E., Stanssens P., Fiers W., Gene, 15, pp. 81-93, (1981)

[4]

DeBoer H.A., Comstock L.J., Vasser M., The tac promoter: a functional hybrid derived from the trp and lac promoters., Proceedings of the National Academy of Sciences, 80, pp. 21-25, (1983)

[5]

Fieschko J., Ritch T., PRODUCTION OF HUMAN ALPHA CONSENSUS INTERFERON IN RECOMBINANT ESCHERICHIA COLI, Chemical Engineering Communications, 45, pp. 229-240, (1986)

[6]

Siegel R., Ryu D.Y., Biotechnol. Bioeng., 27, pp. 28-33, (1985)

[7]

Enberg B., Nordstrom K., J. Bacteriol, 123, pp. 179-186, (1975)

[8]

Wu W.T., Chen K.C., Chiou H.W., Biotechnol. Bioeng., 27, pp. 756-760, (1985)

[9]

Uhlin B.E., Nordstrom K., Mol. Gen. Genet., 165, pp. 176-179, (1978)

[10]

Demain A.L., Solomon N.A., Manual of Industrial Microbiology and Biotechnology, pp. 137-151, (1986)

← 1 2 →