BIOPROCESS DEVELOPMENT TO IMPROVE FOREIGN PROTEIN-PRODUCTION FROM RECOMBINANT STREPTOMYCES

被引:15
作者
DELACRUZ, N
PAYNE, GF
SMITH, JM
COPPELLA, SJ
机构
[1] UNIV MARYLAND,CTR AGR BIOTECHNOL,DEPT CHEM & BIOCHEM ENGN,BALTIMORE,MD 21229
[2] UNIV MARYLAND,CTR MED BIOTECHNOL,BALTIMORE,MD 21229
关键词
D O I
10.1021/bp00016a007
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
Bioprocessing strategies to improve production of the heterologous protein Parathion hydrolase from recombinant Streptomyces lividans were investigated. Initial limitations to increased production were overcome by using large amounts of nutrients and feeding these nutrients throughout the fermentation. Batch addition of such large amounts of nutrients resulted in byproduct acid accumulation. Our data suggest that byproducts resulted from incomplete utilization of peptide medium ingredients and not from an overflow of glucose catabolism. Over extended fed-batch operation, oxygen transfer became limiting and these limitations were overcome by sparging oxygen-enriched gas. When cultivation was continued past about 90 h, we observed that despite nutrient feeding and oxygen enrichment enzyme activities no longer increased. Our results show that during such late cultivation periods the rates of enzyme synthesis and deactivation became balanced. If synthesis is prevented, either by a nutritional limitation or by the addition of the protein synthesis inhibitor chloramphenicol, enzyme activities were observed to decrease. Since deactivation rate constants in these experiments were similar to those observed in cell-free studies, and because extracellular protease activities were not detected in our fermentation, it appears that deactivation results from the inherent instability of the parathion hydrolase enzyme.
引用
收藏
页码:307 / 315
页数:9
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共 55 条
  • [11] THE EFFECT OF DISSOLVED-OXYGEN AND AERATION RATE ON ANTIBIOTIC PRODUCTION OF STREPTOMYCES-FRADIAE
    CHEN, HC
    WILDE, F
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 1991, 37 (06) : 591 - 595
  • [12] GENETIC-ENGINEERING APPROACH TO TOXIC-WASTE MANAGEMENT - CASE-STUDY FOR ORGANOPHOSPHATE WASTE TREATMENT
    COPPELLA, SJ
    DELACRUZ, N
    PAYNE, GF
    POGELL, BM
    SPEEDIE, MK
    KARNS, JS
    SYBERT, EM
    CONNOR, MA
    [J]. BIOTECHNOLOGY PROGRESS, 1990, 6 (01) : 76 - 81
  • [13] CLONING AND CHARACTERIZATION OF A GENE OF STREPTOMYCES-GRISEUS THAT INCREASES PRODUCTION OF EXTRACELLULAR ENZYMES IN SEVERAL SPECIES OF STREPTOMYCES
    DAZA, A
    GIL, JA
    VIGAL, T
    MARTIN, JF
    [J]. MOLECULAR & GENERAL GENETICS, 1990, 222 (2-3): : 384 - 392
  • [14] GLUCOSE-STIMULATED ACIDOGENESIS BY STREPTOMYCES-PEUCETIUS
    DEKLEVA, ML
    STROHL, WR
    [J]. CANADIAN JOURNAL OF MICROBIOLOGY, 1987, 33 (12) : 1129 - 1132
  • [15] DEVELOPMENT OF STREPTOMYCES-AUREOFACIENS IN SUBMERGED CULTURE
    DOSKOCIL, J
    HOSTALEK, Z
    KASPAROVA, J
    ZAJICEK, J
    HEROLD, M
    [J]. JOURNAL OF BIOCHEMICAL AND MICROBIOLOGICAL TECHNOLOGY AND ENGINEERING, 1959, 1 (03): : 261 - 271
  • [16] ENZYME-PRODUCTION BY GENETICALLY ENGINEERED STREPTOMYCES STRAINS - INFLUENCE OF CULTURE CONDITIONS
    ERPICUM, T
    GRANIER, B
    DELCOUR, M
    LENZINI, VM
    NGUYENDISTECHE, M
    DUSART, J
    FRERE, JM
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 1990, 35 (07) : 719 - 726
  • [17] PRODUCT DEACTIVATION IN RECOMBINANT STREPTOMYCES
    GARDNER, AR
    CADMAN, TW
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 1990, 36 (03) : 243 - 251
  • [18] OPTIMIZING THE PRODUCTION OF RECOMBINANT PROTEINS IN MICROORGANISMS
    GEORGIOU, G
    [J]. AICHE JOURNAL, 1988, 34 (08) : 1233 - 1248
  • [19] EXPRESSION OF A THERMOMONOSPORA-FUSCA CELLULASE GENE IN STREPTOMYCES-LIVIDANS AND BACILLUS-SUBTILIS
    GHANGAS, GS
    WILSON, DB
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1987, 53 (07) : 1470 - 1475
  • [20] SYNTHESIS OF BOVINE GROWTH-HORMONE BY STREPTOMYCES-LIVIDANS
    GRAY, G
    SELZER, G
    BUELL, G
    SHAW, P
    ESCANEZ, S
    HOFER, S
    VOEGELI, P
    THOMPSON, CJ
    [J]. GENE, 1984, 32 (1-2) : 21 - 30