Production ofα,α-Trehalose by a Bacterium Isolated from Soil
Production ofα,α-Trehalose by a Bacterium Isolated from Soil
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从土壤中分离的细菌生产α,α-海藻糖
DOI：
10.1080/00021369.1988.10868754
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Satoshi Mikj
中科院分区：
文献类型：
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作者：
S. Iwahara;Satoshi Mikj
在研究由细菌50 B菌株1.2）产生糖苷酶的过程中，我们发现当细菌在含有甘油作为碳源的培养基上生长时，其在培养滤液中积累二糖。本文报道了双糖的分离和鉴定。使细菌在含有酵母提取物Ig、甘油10 g、（NH 4）2S 〇 4 5g、K2 HP 〇 4 Ig、KCl 0.5g、MgS 〇 4·7 H2 〇 0.5g、FeS 〇 4·7 H2 〇 10 mg、MnCl 2·4 H2 〇 5 mg、CaCl 2·2 H2 〇 20 mg和CuSO 4的培养基上生长。5 H2O 1 mg溶于1000 ml去离子水中，pH7.0，在28 ℃下反复振荡5天。培养后，通过离心除去细胞。将上清液（约500 ml）在50 ℃减压下浓缩至最小体积。将浓缩物施加到Bio-Gel P-2柱（3cm × 100cm）上，然后用去离子水洗脱。将二糖部分合并，然后用去离子水依次通过Dowex I X 2（Cl-，I cm X 5 cm）和Dowex 50（H+，1.8 cm X 10 cm）柱。将水浓缩至约Iml。将浓缩物在Bio-Gel P-2上再层析。将滤液减压浓缩至干。因此，获得约150 mg的干燥材料。该化合物在TLC上产生单一斑点，在酸水解时仅产生葡萄糖（表I）。Fab-质谱显示该化合物的分子量为342，为二糖（数据未显示）。该化合物不还原费林试剂，表明它是两个D-葡萄糖残基通过其还原碳原子连接的二糖。如表I所示，该化合物的Rf值和保留时间与真正的α，α-海藻糖相同。该化合物的比旋光度[al~ O=+ 1800（2%水溶液）]与真正的α，α，-海藻糖相同。对该化合物进行1H-NMR光谱分析，光谱数据见图I。HI和HI'的J值分别为3.6和3.7Hz，而其他质子的J值为9.4~ 10.0Hz。H-I和H-I '的低J值表明该化合物具有α，α-键。从COSY光谱，主要质子的归属如图所示。
During the course of a study on the production of glycosidases by a bacterium, the 50 b strain, 1.2) we found that the bacterium accumulated a disaccharide in the culture filtrate when it was grown on a medium containing glycerol as a carbon source. This paper describes the isolation and identification of the disaccharide. The bacterium was grown on a medium containing yeast extract I g, glycerol 10 g,(NH4) 2S04 5 g, K2HP04 I g, KCI 0.5 g, MgS04'7H20 0.5 g, FeS04· 7H20 10 mg, MnCI2· 4H20 5 mg, CaCI2· 2H20 20 mg and CuSO.· 5H20 I mg in 1000mi of deionized water, pH 7.0, for 5 days at 28 C with reciprocal shaking. After the culture, the cells were removed by centrifugation. The supernatant (about 500 ml) was concentrated to a minimum volume under reduced pressure at 50 e. The concentrate was applied to a column of Bio-Gel P-2 (3 cm x 100 cm) and then eluted with deionized water. The disaccharide portions were combined and then passed through columns of Dowex I x 2 (CI-, I cm x 5 cm) and Dowex 50 (H+, 1.8 cmx 10cm) with deionized water successively. The eluate was concentrated to about I ml. The concentrate was rechromatographed on Bio-Gel P-2. The eluate was concentrated to dryness under reduced pressure. Thus, about 150 mg of a dried material was obtained. This compound gave a single spot on TLC and only glucose on acid hydrolysis (Table I). Fab-mass spectrometry showed that the molecular weight of this compound was 342 as a disaccharide (data not shown). This compound did not reduce Fehling's reagent suggesting that it is a disaccharide of two D-glucose residu~ s linked through their reducing carbon atoms. As shown in Table I, the Rf values and retention times of this compound were identical with those of authentic a, a-trehalose. The specific rotation of this compound,[al~ O=+ 1800 (2% in water), was found to be identical with that of authentic a, a,-trehalose. 1 H-NMR spectrometry was performed on this compound and the spectral data are shown in Fig. I. The J values of HI and HI'were found to be 3.6 and 3.7 Hz, respectively, while those of the other protons were 9.4~ 10.0 Hz. The low J values of H-I and H-I'indicate that this compound has an a, a-linkage. From the COSY spectrum, the main protons were assigned as shown in Fig.