caa a22 4500 465824498 CHVBK 20180323112145.0 cr unu---uuuuu 170327e19900801xx s 000 0 eng 10.1007/BF00172543 doi (NATIONALLICENCE)springer-10.1007/BF00172543 Purification and properties of a thermostable pullulanase from Clostridium thermosulfurogenes EM1 which hydrolyses both α-1,6 and α-1,4-glycosidic linkages [Elektronische Daten] [A. Spreinat, G. Antranikian] Summary: A novel thermostable pullulanase, secreted by the thermophilic anaerobic bacterium Clostridium thermosulfurogenes EM1, was purified and characterized. Applying anion exchange chromatography and gel filtration the enzyme was purified 47-fold and had a specific activity of 200 units/mg. The molecular mass of this thermostable enzyme was determined to be 102 000 daltons and consisted of a single subunit. The enzyme was able to attack specifically the α-1,6-glycosidic linkages in pullulan and caused its complete hydrolysis to maltotriose. Surprisingly and unlike the enzyme from Klebsiella pneumoniae, the purified enzyme from this anaerobic thermophile exhibited, in addition to its debranching and pullulanase activity, an α-1,4 hydrolysing activity as well. By the action of this single polypeptide chain various branched and linear polysaccharides were completely converted to two major products, namely maltose and maltotriose. The K m values of this enzyme for pullulan and amylose were determined to be 1.33 mg/ml and 0.38 mg/ml, respectively. This debranching enzyme displays a temperature optimum at 60°-65° C and a pH optimum at 5.5-6.0. The application of this new class of pullulanase (pullulanase type II) in industry will significantly enhance the starch saccharification process. Springer-Verlag, 1990 Spreinat A. Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut Antranikian G. Arbeitsbereich Biotechnologie I, Technische Universität Hamburg-Harburg, Harburger Schlossstrasse 37-39, D-2100, Hamburg 90, Federal Republic of Germany aut Applied Microbiology and Biotechnology Springer-Verlag 33/5(1990-08-01), 511-518 0175-7598 33:5<511 1990 33 253 https://doi.org/10.1007/BF00172543 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00172543 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Spreinat A. Institut für Mikrobiologie der Georg-August-Universität Göttingen, Grisebachstrasse 8, D-3400, Göttingen, Federal Republic of Germany aut NATIONALLICENCE 700 1- Antranikian G. Arbeitsbereich Biotechnologie I, Technische Universität Hamburg-Harburg, Harburger Schlossstrasse 37-39, D-2100, Hamburg 90, Federal Republic of Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 33/5(1990-08-01), 511-518 0175-7598 33:5<511 1990 33 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer