传统奶酪中产转糖基活性β-半乳糖苷酶乳酸菌的筛选鉴定及其合成低聚半乳糖条件的研究

摘要/Abstract

摘要： （目的）分离筛选产转糖基活性β-半乳糖苷酶的乳酸菌新菌株，为酶法高效合成低聚半乳糖（galacto-oligosaccharides，GOS）提供新的酶源。（方法）以乳糖为唯一碳源，碳酸钙溶钙圈和添加 5-溴-4-氯-3-吲哚-β-D-半乳糖苷（X-Gal）的乳酸细菌培养基（MRS）筛选平板进行初筛，以产酶菌株粗酶液催化乳糖反应产物的TLC分析复筛，获得产转糖基活性β-半乳糖苷酶的乳酸菌。结合其形态学、生理生化特征及16S rRNA序列同源性分析对产转糖基活性β-半乳糖苷酶菌株进行鉴定。单因素实验确定最佳产酶条件和转糖基反应条件，TLC结合HPLC分析转糖基反应产物各组分含量。（结果）筛选获得产转糖基活性β-半乳糖苷酶的菌株6株，其中Lactobacillus plantarum YLBGNL-S7所产β-半乳糖苷酶的转糖基活性最强。单因素实验表明，在温度为50 °C、pH6.0、乳糖浓度30% (w/v)条件下，反应4 h，GOS的得率可达43.40% (w/w)，其中转移二糖和转移三糖含量分别为18.29% (w/w)和12.95% (w/w)。

关键词: β-半乳糖苷酶, 转糖基活性, 低聚半乳糖（GOS）, 植物乳杆菌, 奶酪

Abstract: (Purpose) This study is aiming to isolate and screen lactic acid bacteria strain producing β-galactosidase with high transglycosylation activity in order to provide novel source of β-galactosidase for efficient synthesis of galacto-oligosaccharides (GOS). (Method) Lactic acid bacteria strains were isolated from traditional cheese samples made by the local minority collected in Yili, Xinjiang Province, and screened using lactose as the sole carbon source and CaCO3 dissolving ring as an indicator. Then MRS plate supplemented with X-Gal was used to screen lactic acid bacteria strains producing β-galactosidase. Finally, strains producing β-galactosidases with transglycosylation activity were screened by TLC analysis. The isolated lactic acid bacteria strains were identified by physiological and biochemical characteristics, together with 16S rRNA gene sequence analysis. The optimal temperature, pH and lactose concentration for the synthesis of GOS using lactose as substrate were investigated, and the amount of oligosaccharides in the sample under optimal reaction condition were quantified by TLC together with HPLC using Hi-Plex Na Column and RID-10A detector. (Results) Six lactic acid bacteria strains producing β-galactosidases with transglycosylation activity were isolated, among which the β-galactosidase produced by Lactobacillus plantarum YLBGNL-S7 showed the highest transglycosylation activity. An initial lactose concentration of 30% (w/v), a reaction condition of 50 °C, pH6.0 and a reaction time of 4 h were the optimum conditions for GOS synthesis of the β-galactosidase produced by L. plantarum YLBGNL-S7, which resulted in a GOS yield of 43.40% (w/w). The GOS mixture was composed of 18.29% (w/w) of transglyco-diaccharides and 12.95% (w/w) of tri-saccharides. These results demonstrated that L. plantarum YLBGNL-S7 screened in this study was a novel promising source of β-galactosidase with good potential for prebiotic GOS synthesis.

Key words: β-galactosidase, transglycosylation activity, galacto-oligosaccharides (GOS), Lactobacillus plantarum, cheese

中图分类号:

Q815

王丽军牟元珍关波万丽云张艳胡有贞倪永清. 传统奶酪中产转糖基活性β-半乳糖苷酶乳酸菌的筛选鉴定及其合成低聚半乳糖条件的研究[J]. 食品科学, 0, (): 0-0.

Li-Jun WANG Bo Guan Youzhen Hu. Screening and identification of lactic acid bacteria strain producing β-galactosidases with transglycosylation activity from traditional cheese, and the condition optimization for galacto-oligosaccharides synthesis[J]. FOOD SCIENCE, 0, (): 0-0.

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