Beta-glucoside permeases and phospho beta-glucosidases in Aerobacter aerogenes: relationship with cryptic phospho beta-glucosidases in Enterobacteriaceae.
Beta-glucoside permeases and phospho beta-glucosidases in Aerobacter aerogenes: relationship with cryptic phospho beta-glucosidases in Enterobacteriaceae.
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产气杆菌中的 β-葡萄糖苷渗透酶和磷酸 β-葡萄糖苷酶：与肠杆菌科中隐秘的磷酸 β-葡萄糖苷酶的关系。
DOI：
10.1073/pnas.59.1.285
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发表时间：
1968
影响因子：
11.1
通讯作者：
I. Schenkein
中科院分区：
文献类型：
--
作者：
S. Schaefler;I. Schenkein
材料和方法。-细菌菌株：菌株A. aerogenes A1 是我们部门的收集菌株。 Tanaka 和 Lin 描述的菌株 A. aerogenes 2002（gua-、arg-）及其衍生物 2050 和 2070 7 是从 E. Lin 博士获得的。菌株 K12 j#-gl- 及其 j3-gl+ 突变体先前已被描述。 5 菌株 E. coli Hu（纤维二糖+、水杨苷-、熊果苷-）来自 Dr. F.0rskov。柠檬酸杆菌菌株 B 购自 F. Kauffmann 博士，菌株 532 购自 E. Ewing 博士。鼠伤寒沙门氏菌菌株 489（ile-、pur-、met-）由 G. Dubnau 博士提供；从该菌株中获得了两个自发纤维二糖发酵突变体（C1+和C2+）。 S,+，C1+ i 的自发突变体，此外也发酵水杨苷和熊果苷，并且全细胞组成型分裂PNP-glu。培养基：对于酶和渗透酶测定，细菌在补充有0.2％酵母提取物、0.5％琥珀酸钠和所需生长因子的培养基A、8中生长，培养基AY。 5 对于无细胞提取物，细胞通过在含有 LB-1% 酵母提取物、2% 胰蛋白胨 (Difco)、0.1% NaCl 的培养基中通气培养过夜，并用 NaOH 调节至 pH 7.0。 β3-葡萄糖苷的主动摄取：C'-噻吩 B-葡萄糖苷 (TPG) 的主动摄取是在之前针对大肠杆菌 5 描述的条件下测定的，但不存在氯霉素。使用 2X 10-4 M 标记化合物和 10-2 M 冷化合物进行“追逐”实验。酶测定：如前所述，通过测量糖苷配基的释放来测定对硝基苯基、B-葡萄糖苷 (PHP-glu)、邻硝基苯基、-葡萄糖苷 (ONP-glu)、苯基、B-葡萄糖苷和水杨苷的裂解 5。一单位酶对应于1 毫摩尔苷元/分钟的磷酸酶测定是在 0.15 毫升体积中进行的，其中含有： 5 毫摩尔的测试磷酸盐。
Materials and Methods.-Bacterial strains: The strain A. aerogenes A1 is a collection strain from our Department. The strain A. aerogenes 2002 (gua-, arg-) and its derivatives 2050 and 2070, described by Tanaka and Lin, 7 were obtained from Dr. E. Lin. The strain E. coli K12 j#-gl-and its j3-gl+ mutants have been described previously. 5 The strain E. coli Hu (cellobiose+, salicin-, arbutin-) was received fromDr. F. 0rskov. The Citrobacter strain B was received from Dr. F. Kauffmann and strain 532 from Dr. E. Ewing. The S. typhimurium strain 489 (ile-, pur-, met-) was received from Dr. G. Dubnau; from this strain two spontaneous cellobiose-fermenting mutants were obtained (C1+ and C2+). S,+, a spontaneous mutant of C1+ i in addition also ferments salicin and arbutin, and whole cells split PNP-glu constitutively. Culture media: For enzyme and permease determinations the bacteria were grown in medium A, 8 supplemented with 0.2% yeast extract, 0.5% Na succinate, and the required growth factors-medium AY. 5 For cell-free extracts the cells were grown overnight by aeration in medium LB-1% yeast extract, 2% tryptone (Difco), 0.1% NaCl, adjusted to pH 7.0 with NaOH. Active uptake of j3-glucosides: The active uptake of C"-thiophenyl B-glucoside (TPG) was determined under the conditionspreviously described forE. coli, 5 but in the absence of chloramphenicol." Chasing" experiments were performed with 2X 10-4 M of the labeled compound and 10-2 M of the cold compound. Enzyme determinations: The determination of the splitting of p-nitrophenyl, B-glucoside (PHP-glu), o-nitrophenyl,-glucoside (ONP-glu), phenyl, B-glucoside, and salicin was made, as previously described, 5 by measuring the liberation of the aglycone. One unit of enzyme corresponds to the liberation of 1 mnumole aglycone/min. Phosphatase determinations were made in a volume of 0.15 ml containing: 5 jmoles of the tested phosphate