Heterologous production of streptokinase as a secretory form in Streptomyces lividans and nonsecretory form in Escherichia coli. 2010

Mi-Ran Kim, and Yong-Hoon Choeng, and Won-Jae Chi, and Dae-Kyung Kang, and Soon Kwang Hong
Department of Biological Science, Myongji University, Yongin 449-728, Korea.

The skc gene encoding streptokinase (SK), with a molecular weight of approximately 47.4 kDa, was cloned from Streptococcus eouisimilis ATCC9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the sprT promoter was used in the S. lividans TK24 host, the SK protein corresponding to 47.4 kDa was detected with a smaller hydrolyzed protein (44 kDa), implying posttranslational hydrolysis occurred as reported in other expression systems. Casein/plasminogen plate assay revealed that plasmid construct with the signal peptide of SK was superior to that with the signal peptide of sprT in SK production. The maximum productivity of SK was calculated as less than 0.25 unit/ml of the culture broth, which was similar level to those from other expression systems hiring ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(DE3)pLys under control of T7 promoter, relatively large amount of SK was expressed in soluble form without hydrolyzed protein. The SK activity produced by E. coli/pET28a-T7pSKm was more than 2 units/ml of culture even though about half of the expressed protein formed inactive inclusion body.

UI MeSH Term Description Entries
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D001426 Bacterial Proteins Proteins found in any species of bacterium. Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D013291 Streptococcus A genus of gram-positive, coccoid bacteria whose organisms occur in pairs or chains. No endospores are produced. Many species exist as commensals or parasites on man or animals with some being highly pathogenic. A few species are saprophytes and occur in the natural environment.
D013300 Streptokinase Streptococcal fibrinolysin . An enzyme produced by hemolytic streptococci. It hydrolyzes amide linkages and serves as an activator of plasminogen. It is used in thrombolytic therapy and is used also in mixtures with streptodornase (STREPTODORNASE AND STREPTOKINASE). EC 3.4.-. Avelizin,Awelysin,Celiase,Distreptase,Kabikinase,Kabivitrum,Streptase,Streptodecase
D015870 Gene Expression The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION. Expression, Gene,Expressions, Gene,Gene Expressions
D048170 Streptomyces lividans An actinomycete used for production of commercial ANTIBIOTICS and as a host for gene cloning.
D021381 Protein Transport The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport. Cellular Protein Targeting,Gated Protein Transport,Protein Targeting, Cellular,Protein Translocation,Transmembrane Protein Transport,Vesicular Protein Transport,Protein Localization Processes, Cellular,Protein Sorting,Protein Trafficking,Protein Transport, Gated,Protein Transport, Transmembrane,Protein Transport, Vesicular,Traffickings, Protein
D021382 Protein Sorting Signals Amino acid sequences found in transported proteins that selectively guide the distribution of the proteins to specific cellular compartments. Leader Signal Peptides,Leader Peptide,Leader Sequences, Peptide,Peptide Leader Sequences,Peptide Signal Sequences,Signal Peptide,Signal Peptides,Signal Sequence, Peptide,Signal Sequences,Signal Sequences, Peptide,Leader Peptides,Leader Sequence, Peptide,Leader Signal Peptide,Peptide Leader Sequence,Peptide Signal Sequence,Peptide, Leader,Peptide, Leader Signal,Peptide, Signal,Peptides, Leader,Peptides, Leader Signal,Peptides, Signal,Protein Sorting Signal,Sequence, Peptide Leader,Sequence, Peptide Signal,Sequence, Signal,Sequences, Peptide Leader,Sequences, Peptide Signal,Sequences, Signal,Signal Peptide, Leader,Signal Peptides, Leader,Signal Sequence,Signal, Protein Sorting,Signals, Protein Sorting,Sorting Signal, Protein,Sorting Signals, Protein

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