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产β-半乳糖苷酶乳酸菌的筛选及酶法合成低聚半乳糖的GC-MS分析
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作者简介:张宏志(1985-),男,博士,助理研究员,主要从事功能性低聚糖和多糖的研究; 通讯作者:马艳弘(1972-),女,博士,副研究员,主要从事生物技术与功能食品研究

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江苏省农业科技自主创新资金自由探索项目(CX(14)5059);江苏省科技支撑计划(农业)(BE2014347)


Screening of -Galactosidase-producing Lactic Acid Bacteria and Analysis of the Enzymatic Synthesis of Galactooligosaccharides by GC-MS
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    摘要:

    从分离自13种中国传统发酵食品的148株乳酸菌中筛选产高转糖基活性β-半乳糖苷酶的菌株。实验采用X-Gal平板初筛、薄层层析(TLC)复筛、气相色谱(GC)定量的方法,得到一株产β-半乳糖苷酶转糖基活性最高的菌株,并以乳糖为单底物,利用该高转糖基活性β-半乳糖苷酶酶法合成低聚半乳糖(GOS),并采用GC-MS的方法对其各个组分进行鉴定。研究结果表明,菌株70810所产β-半乳糖苷酶转糖基活性最高,其合成GOS产率达到39.23%(m/m);该菌株为实验室已鉴定乳酸菌,为植物乳杆菌(HQ259238) (Lactobacillus plantarum);酶法合成的GOS产物鉴定为9种低聚二糖和3种低聚三糖,主要结构多为β(1→6)和β(1→3)糖苷键。菌株70810来源于泡菜,安全性好,所产β-半乳糖苷酶转糖基活性最高,且可不经纯化直接利用,在食品与乳品加工等方面将具有很好的应用前景。?

    Abstract:

    Lactic acid bacteria (LAB) strains producing ?-galactosidases with high transgalactosylation activity were screened from 148 LAB strains found in 13 kinds of traditional Chinese fermented foods. The strain 70810 produced ?-galactosidases with the highest transgalactosylation activity and was isolated using a 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-Gal) plate for the primary screening, thin layer chromatography (TLC) for the secondary screening, and gas chromatography (GC) for quantitation. Lactose was used as a single substrate, the synthesis of galactooligosaccharides (GOS) was catalyzed by ?-galactosidases with high transgalactosylation activity, and gas chromatography-mass spectrometry (GC-MS) was used to identify each component. The results showed that the ?-galactosidases produced by strain 70810 had the highest transgalactosylation activity, and the yield of GOS was 39.23 % (m/m). This LAB strain was identified as Lactobacillus plantarum (HQ259238). Among the GOS products derived from enzymatic synthesis, there were nine disaccharides and three trisaccharides and the main structures contained ?-1,6 and ?-1,3 glycosidic bonds. Because strain 70810 was isolated from Chinese Paocai, it is safe. The resulting ?-galactosidases have the highest transgalactosylation activity and can be directly used without separation and purification. Therefore, this strain has potential applications in food and dairy industries.

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张宏志,马艳弘,黄开红,李亚辉.产β-半乳糖苷酶乳酸菌的筛选及酶法合成低聚半乳糖的GC-MS分析[J].现代食品科技,2015,31(12):355-361.

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  • 收稿日期:2015-06-04
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  • 在线发布日期: 2015-12-30
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