食品科学 ›› 2007, Vol. 28 ›› Issue (5): 241-244.

• 生物工程 • 上一篇    下一篇

转糖基β-半乳糖苷酶生产含低聚半乳糖的低乳糖牛奶

 李正义, 肖敏, 卢丽丽, 李玉梅   

  1. 山东大学微生物技术国家重点实验室; 山东大学微生物技术国家重点实验室 山东济南250100; 山东济南250100;
  • 出版日期:2007-05-15 发布日期:2011-12-31

Production of Low-lactose Milk Containing Galactooligosaccharides withβ-Galactosidase-catalyzed Transgalactosylation from Enterobacter sp.B5

 LI  Zheng-Yi, XIAO  Min, LU  Li-Li, LI  Yu-Mei   

  1. State Key Laboratory of Microbial Technology,Shandong University,Jinan 250100,China
  • Online:2007-05-15 Published:2011-12-31

摘要: 研究利用具有转糖基活性的β-半乳糖苷酶生产含低聚半乳糖的低乳糖牛奶。含低聚半乳糖的低乳糖牛奶既能解决乳糖不耐症问题,同时还在牛奶中增添了低聚半乳糖双歧因子。从Enterobacter sp.B5的无细胞抽提液中,利用硫酸铵沉淀制备具有转糖基活性的β-半乳糖苷酶,作用鲜牛奶生成含低聚半乳糖的低乳糖牛奶。利用薄层层析、高压液相色谱技术和软件NIH ImageJ 1.36分析了反应产物中的糖组分(葡萄糖、半乳糖、乳糖和低聚糖(包括转移二糖和三糖))。结果显示酶作用后的鲜牛奶中大约70%的乳糖转化为葡萄糖和半乳糖,至少10%的乳糖通过转糖基反应生成低聚半乳糖。在酶浓度为3U/ml,50℃酶解4h的反应条件下,获得含低聚半乳糖的低乳糖牛奶的糖组成为0.59%低聚半乳糖(含0.37%转移二糖和0.22%三糖),1.33%乳糖,2.83%单糖。本研究利用软件NIH ImageJ 1.36定量分析样品糖组分的TLC结果,可以避免乳糖的同分异构体-转移二糖在高压液相色谱结果中因无法分离而造成的漏检。

关键词: &beta, -半乳糖苷酶, 转糖基作用, 低聚半乳糖, 低乳糖牛奶

Abstract: Production of low-lactose milk containing galactooligosaccharides was studied withβ-Galactosidase by transgalactosylation activity from Enterobacter sp.B5.In this way it can ease lactose intolerance.Meanwhile,the galactooligosaccharides as bifidus factor can promote the growth of bifidobacterium.The fresh cow milk was then directly treated withβ-galactosidase of transgalactosylation activity derived from Enterobacter sp.B5.The types and amounts of total sugars of processed milk were analyzed by thin layer chromatography(TLC),high pressure liquid chromatography(HPLC)and the software NIH ImageJ 1.36.About 70% of lactose contained in the cow milk was hydrolyzed to glucose and galactose,while at least 10% of lactose in the cow milk was changed into galactoofigosaccharides by the transgalactosidafion reaction.When the cow milk was treated withβ-galactosidase at an enzyme concentration of 3 U/ml at 50℃in 4 hours,a processed milk was obtained containing 0.59% of galactooligosaccharides including 0.37% of transgalactosylated disaccharides and 0.22% trisaccharide, 1.33% of lactose,and 2.83% of monosaccharide.The use of public-domain NIH ImageJ software for the quantitation of scanned images of thin-layer chromatography can help examine the transgalactosylated disaccharide(the isomer of lactose),which would otherwise not be detected in HPLC chromatogram comparatively.

Key words: &beta, -galactosidase; transgalactosidation; galactooligosaccharides; low-lactose milk;