Journal Article Supplementary studies on an extracellular proteinase of Prevotella intermedia: formation and some enzymatic roperties
Prevotella intermedia の細胞外タンパク分解酵素の補充的研究:産生と酵素性状

KISO, AKIKO  ,  MATSUO, KAZUKO  ,  SHIBATA, YUKINAGA  ,  HASEGAWA, HIROMASA  ,  YOSHIDA, AKIHIRO  ,  FUJIMURA, SETSUO  ,  木曽, 有紀子  ,  松尾, 和子  ,  柴田, 幸永  ,  長谷川, 博雅  ,  吉田, 明弘  ,  藤村, 節夫

41 ( 1 )  , pp.1 - 6 , 2015-06-30 , 松本歯科大学学会
ISSN:2188-7233
NCID:AA12684554
Description
Formation and some enzymatic properties of a proteinase of Prevotella intermedia ATCC 2₅611 were examined using Azocoll, a solid chromogenic non–specific proteinase substrate. Anaerobic globe box supported better growth of cell and production of proteinase than AnaeroPak. Nearly all proteinase activity was found in the extracellular culture fluid, but not in the cell extract and the envelope. Production of proteinase started immediately after inoculation of the bacteria to the medium and continued until the middle stage of the stationary phase of the growth. Proteinase production was strongly inhibited by addition of glucose or fructose to the medium, moderately by sucrose and weakly by galactose. This proteinase was inactivated entirely by heating at 60℃ for 20 min. Ethyleneglycol bis (β –aminoethyl ether)–N,N,N,N ʼ–tetraacetate (EGTA), Zn2+ and Cu2+ completely inhibited the proteolysis. No effect on the activity was observed by Antipain, SDS, N–tosyl–L–phenylalanyl oromethyl ketone (TPCK), Ca2+ and Mg2+.
Prevotella intermedia ATCC 2₅611のタンパク分解酵素の産生および性状についてアゾコルを酵素基質として調べた.嫌気ボックスとアネロパックで培養したところ,前者の方が増殖も酵素の産生は優れていた.酵素活性のほとんどは菌体外の培養上清に認められ,無細胞抽出液とエンベロープには検出されなかった.酵素産生は培養開始直後から開始され,定常期中ごろまで続いた.また,この酵素の産生は培地にグルコースやフルクトースを加えると強く抑制され,シュクロースでは中等度の,ガラクトースでは弱い抑制が観察された.温度感受性テストにより,60℃,20分加熱で失活し,EGTA, Zn2+,Cu2+ で活性は阻害されるが,アンチパイン,SDS,TPCK,Ca2+,Mg2+ による影響はなかった.
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