geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Extracellular secretion of pullulanase is unaffected by minor sequence changes but is usually prevented by adding reporter proteins to its N- or C-terminal end


, : Extracellular secretion of pullulanase is unaffected by minor sequence changes but is usually prevented by adding reporter proteins to its N- or C-terminal end. Journal of Bacteriology 177(18): 5238-5246

Linker insertions in the pullulanase structural gene (pulA) were examined for their effects on pullulanase activity and cell surface localization in Escherichia coli carrying the cognate secretion genes from Klebsiella oxytoca. Of the 23 insertions, 11 abolished pullulanase activity but none were found to prevent secretion. To see whether more drastic changes affected secretion, we fused up to five reporter proteins (E. coli periplasmic alkaline phosphatase, E. coli periplasmic maltose-binding protein, periplasmic TEM beta-lactamase, Erwinia chrysanthemi extracellular endoglucanase Z, and Bacillus subtilis extracellular levansucrase) to three different positions in the pullulanase polypeptide: close to the N terminus of the mature protein, at the C terminus of the protein, or at the C terminus of a truncated pullulanase variant lacking the last 256 amino acids. Only 3 of the 13 different hybrids were efficiently secreted: 2 in which beta-lactamase was fused to the C terminus of full-length or truncated pullulanase and 1 in which maltose-binding protein was fused close to the N terminus of pullulanase. Affinity-purified endoglucanase-pullulanase and pullulanase-endoglucanase hybrids exhibited apparently normal levels of pullulanase activity, indicating that the conformation of the pullulanase segment of the hybrid had not been dramatically altered by the presence of the reporter. However, pullulanase-endoglucanase hybrids were secreted efficiently if the endoglucanase component comprised only the 60-amino-acid, C-terminal cellulose-binding domain, suggesting that at least one factor limiting hybrid protein secretion might be the size of the reporter.

US$29.90

PMID: 7665512


Other references

Eveleigh E.S.; Chant D.A., 1982: Experimental studies on acarine predator prey interactions the effects of predator density on immature survival adult fecundity and emigration rates and the numerical response to prey density acarina phytoseiidae. Laboratory experiments revealed that increasing the density of the immature instars of Phytoseiulus persimilis had little effect on their survival rate. Due to the high prey requirements of the immature instars of Amblyseius degenerans and the ten...

Ancrenaz, M.; Calaque, R.; Lackman Ancrenaz, I., 2004: Orangutan nesting behavior in disturbed forest of Sabah, Malaysia: implications for nest census. High concentrations of orangutans remain in the multiple-use forests of the Lower Kinabatangan, Sabah, Malaysia. Compared to primary forest, the habitat is highly fragmented, characterized by a low tree density (332 stems/ha), small tree size (83....

Mehta, A.B.; Biswas, A.K.; Pinto, I.J.; Sheth, U.K., 1967: Clinical assessment of the new antihypertensive Compound--1029 (Vatensol)--Guanoclor. Indian Heart Journal 19(4): 322-328

V.B.Brin, 1977: Changes in the systemic hemodynamics produced by vasopressin in dogs during individual development. Indices of the central hemodynamics were studied in puppies aged 1822 days and 23 months and in dogs aged 35 years after intravenous injection of synthetic vasopressin (0.8 unit/kg body weight). The pressor effect was strongest in the adult dogs a...

Schmidegg Oskar, 1954: Achsen- und Flaechengefuege beiderseits des Silltalbruches zwischen Innsbruck und Matrei. Description of the fabric of quartz phyllites on the eastern side and gneisses on the western side of the Sill valley between Innsbruck and Matrei, Austria.

Hutchison, R., 2007: Opioids: the role in headache pharmacotherapy. Optimal acute opioid management involves a continual awareness of the potential for development of MOH. Both the clinician and patient should be aware of the IHS MOH diagnostic criteria. Prophylactic medications should be initiated for patients ha...

Schey S.A.; Cooper J.; Summerhayes M., 1992: The handfoot syndrome occurring with chronic administration of etoposide. European Journal of Haematology 48(2): 118-119

Gendre, J.P.; Hardouin, J.P.; Marche, C.; Debray, C., 1978: Ulcero-proliferative or granulomatous colitis of amoebic origin. Presentation of three cases. Three cases of amoeboma, in the right colon, the transverse colon and the rectum, are reported. 2 patients had had dysentery while in Indo-China many years previously. Diagnosis is complicated by the similarity of this form of amoebiasis to other...

Leonardi, A.; Briggs, R.M.; Bloch, K.J.; Allansmith, M.R., 1990: Clinical patterns of ocular anaphylaxis in guinea pigs passively sensitized with IgG1 antibody. A model of topically induced ocular anaphylaxis in guinea pigs was developed. A guinea pig anti-dinitrophenyl (DNP) bovine .gamma.-globulin antiserum rich in IgG1 antibodies was injected subconjunctivally; various serum dilutions and latent period...

Thompson, A R., 2002: Habitat-use patterns of the fierce shrimpgoby (Ctenogobiops feroculus) at multiple spatilal scales: a geospatial approach. Unknown