TY - JOUR
T1 - Role of lipid modification on a starch‐debranching enzyme, Klebsieila pullulanase
T2 - comparison of properties of lipid‐modified and unmodified pullulanases
AU - Yamashita, M.
AU - Nakagawa, A.
AU - Katsuragi, N.
AU - Murooka, Y.
PY - 1992/2
Y1 - 1992/2
N2 - Klebsiella pullulanase is a lipoprotein synthesized as a precursor with a signal peptide, which is processed by lipoprotein signal peptidase. To clarify the role of lipid modification of pullulanase, we purified lipid‐modified wild‐type and the unmodified (mutant) pullulanases and compared their properties. The Km and Vmax values of both pullulanases for pullulan were the same. The optimal pH and temperature, the stabilities over pH and temperature ranges, the specificity of substrates, and the patterns of inhibition of the lipid‐modified and unmodified pullulanases were also the same. However, we found that the wild‐type pullulanase formed trimers whereas the unmodified enzyme did not, and that the migrations of the two enzymes on sodium dodecyl sulphate/electrophoresis were different when the samples were applied on the gel without heating. The results presented in this paper and in previous work show that the correct processing and translocation of pullulanase in K. aerogenes require modification of lipid. However, the enzymatic properties and physical stabilities of pullulanase were not affected by the lipid modification.
AB - Klebsiella pullulanase is a lipoprotein synthesized as a precursor with a signal peptide, which is processed by lipoprotein signal peptidase. To clarify the role of lipid modification of pullulanase, we purified lipid‐modified wild‐type and the unmodified (mutant) pullulanases and compared their properties. The Km and Vmax values of both pullulanases for pullulan were the same. The optimal pH and temperature, the stabilities over pH and temperature ranges, the specificity of substrates, and the patterns of inhibition of the lipid‐modified and unmodified pullulanases were also the same. However, we found that the wild‐type pullulanase formed trimers whereas the unmodified enzyme did not, and that the migrations of the two enzymes on sodium dodecyl sulphate/electrophoresis were different when the samples were applied on the gel without heating. The results presented in this paper and in previous work show that the correct processing and translocation of pullulanase in K. aerogenes require modification of lipid. However, the enzymatic properties and physical stabilities of pullulanase were not affected by the lipid modification.
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U2 - 10.1111/j.1365-2958.1992.tb01482.x
DO - 10.1111/j.1365-2958.1992.tb01482.x
M3 - Article
C2 - 1552852
AN - SCOPUS:0026529109
VL - 6
SP - 389
EP - 394
JO - Molecular Microbiology
JF - Molecular Microbiology
SN - 0950-382X
IS - 3
ER -