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益生菌生物药物是指通过口服表达药用多肽(蛋白)的重组益生菌活细胞达到治疗疾病的新型口服给药系统。为了构建一种能有效防治2型糖尿病的酵母生物药物,文章首先构建了酿酒酵母(S.cerevisiae)整合型表达载体p NK1-PGK,并且通过绿色荧光蛋白(GFP)证明其表达功能正常,利用该载体将10?GLP-1(Glucagon-like peptide-1)基因转化到酿酒酵母INVSc1中,通过营养缺陷型和Western blotting成功筛选出表达10?GLP-1的长效促胰岛素降糖酵母(Long-acting GLP-1 hypoglycemic yeast,LHY)。该酵母生长迅速,外源基因10?GLP-1表达稳定,表达量达到1.56 mg/g细胞湿重。通过链脲佐菌素和高脂高糖饮食联合诱导的方法构建了2型糖尿病小鼠模型,用LHY对其进行口服灌胃治疗,证明LHY具有较好疗效,明显降低血糖水平。
Probiotics Biopharmaceuticals refers to a novel oral delivery system for the treatment of diseases by the administration of recombinant probiotic live cells orally expressing a pharmaceutical polypeptide (protein). In order to construct a yeast biopharmaceutical that can effectively prevent and treat type 2 diabetes, we first constructed an integrated expression vector p NK1-PGK of S. cerevisiae and confirmed its expression by green fluorescence protein (GFP) The vector was used to transform 10? GLP-1 (peptide-1) gene into Saccharomyces cerevisiae INVSc1, and the long-acting insulinotropic hypoglycemic yeast expressing 10? GLP-1 was successfully selected by auxotrophic and Western blotting -acting GLP-1 hypoglycemic yeast, LHY). The yeast grew rapidly and the expression of exogenous gene 10? GLP-1 was stable, reaching 1.56 mg / g cell wet weight. The model of type 2 diabetes mice was established by the combination of streptozotocin and high-fat and high-sugar diet, and oral administration of LHY to LHY had good curative effect and significantly decreased blood glucose level.