Lanz W W, Williams P P
J Bacteriol. 1973 Mar;113(3):1170-6. doi: 10.1128/jb.113.3.1170-1176.1973.
An obligately anaerobic ruminal bacterial isolate was selected from 18 tributyrin-degrading isolates and identified as Butyrivibrio fibrisolvens strain 53. The culture in late exponential phase contained enzymes which could be released by sonic disruption. These enzymes degraded substrates at a rate in the order 1-naphthyl acetate (NA) > 1-naphthyl butyrate > 1-naphthyl propionate but did not degrade 1-naphthyl palmitate or 1-naphthyl phosphate. The enzymes on NA were neither stimulated nor inhibited by CoCl(2), MgCl(2), and MnCl (each varied from 10(-6) to 10(-4) M). CaCl at 10(-3) M stimulated esterase activity by 16%. Aliphatic substrates were hydrolyzed at a rate in the order triacetin > tributyrin > tripropionin, and ethyl acetate > ethyl formate. Similarly, aromatic fluorescein diesters were degraded at a rate in the order acetyl > propionyl > caproyl > butyryl > capryl > lauryl. Polyacrylamide gel electrophoretic zymograms indicated that the enzyme composite contained cathodally migrating bands. By column chromatography, these enzymes were separated into six NA-degrading fractions. Fraction V contained an esterase which had an optimal temperature of 39 C, a K(m) of 7.6 x 10(-4) on NA, and a molecular weight of about 66,000. This enzyme was inhibited by paraoxon (41%, 10(-4) M), eserine (17%, 10(-2) M), NaF (17%, 10(-2) M), and diisopropyl fluorophosphate (62%, 10(-4) M) but not by 1-naphthyl N-methyl carbamate at 8.4 x 10(-4) M.
从18株能降解三丁酸甘油酯的分离菌株中挑选出一株严格厌氧的瘤胃细菌分离株，并鉴定为溶纤维丁酸弧菌53株。对数生长后期的培养物中含有可通过超声破碎释放的酶。这些酶降解底物的速率顺序为：1-萘基乙酸酯（NA）＞1-萘基丁酸酯＞1-萘基丙酸酯，但不降解1-萘基棕榈酸酯或1-萘基磷酸酯。NA上的酶不受CoCl₂、MgCl₂和MnCl₂（浓度均在10⁻⁶至10⁻⁴M之间变化）的刺激或抑制。10⁻³M的CaCl₂使酯酶活性提高了16%。脂肪族底物的水解速率顺序为：三醋精＞三丁酸甘油酯＞三丙酸甘油酯，以及乙酸乙酯＞甲酸乙酯。同样，芳香族荧光素二酯的降解速率顺序为：乙酰基＞丙酰基＞己酰基＞丁酰基＞辛酰基＞月桂酰基。聚丙烯酰胺凝胶电泳酶谱表明，酶复合物含有向阴极迁移的条带。通过柱色谱法，这些酶被分离成六个降解NA的组分。组分V含有一种酯酶，其最适温度为39℃，对NA的Kₘ为7.6×10⁻⁴，分子量约为66,000。该酶受到对氧磷（41%，10⁻⁴M）、毒扁豆碱（17%，10⁻²M）、NaF（17%，10⁻²M）和二异丙基氟磷酸酯（62%，10⁻⁴M）的抑制，但不受8.4×10⁻⁴M的1-萘基N-甲基氨基甲酸酯的抑制。
