1株酸性α-淀粉酶产生菌的鉴定及发酵条件优化  

Identification of an Acidic Bacterial Strain Producing α-amylase and the Optimization of Its Fermentation Condition

作　　者：安弋[1] 刘新育[1] 芦鹏[1] 刘亮伟[1] 陈红歌[1] 

机构地区：[1]河南农业大学生命科学学院,河南郑州450002

出　　处：《安徽农业科学》2010年第4期1709-1711,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]研究酸性α-淀粉酶高产菌株B-5的发酵特性并对B-5菌株进行细菌鉴定,为进一步挖掘该菌株的应用价值奠定基础。[方法]以B-5菌株为研究对象,通过菌体形态、菌落特征、生理生化特性以及16Sr DNA同源性分析对其进行菌种鉴定。以单因素法和正交试验法对培养基进行优化,确定B-5菌株的最适发酵条件。[结果]通过传统细菌鉴定方法和分子生物学方法将B-5菌株初步鉴定为芽孢杆菌(Bacillus sp.)。以可溶性淀粉、玉米粉、小麦粉作为碳源时,都有利于B-5菌株产酶,从工业化生产降低成本的角度考虑,选用玉米粉作为发酵产酶的碳源。而以胰蛋白胨作为氮源时产酶活力最高,故选用胰蛋白胨作为发酵产酶的氮源。正交试验结果表明,2%玉米粉和2%胰蛋白胨的培养基组成最有利于发酵产酶,酶活为74.6U/ml。B-5菌株在60h时,发酵液中酶活力达到最高,随着时间的延长,酶活力逐渐降低。[结论]B-5菌株虽然产酶活力还有限,尚不能满足工业化生产的需要,但该菌株的酸性α-淀粉酶性质优异,通过对其菌种的鉴定则可以方便地获取该酸性α-淀粉酶的基因以尝试酸性α-淀粉酶的工程菌生产。[Objective]The basis of the value of an acidic bacteria strain-B-5 of α-amylase-producing was further laid through the study on its high-yielding fermentation characteristics and its identification.[Method]The strain-B-5 being taken as the research object,its identification was done through the morphology,colony characteristics,physiological and biochemical characteristics and the analysis of 16S rDNA homology.The best fermentation condition of the strain was determined based on the optimum culture medium through the method of single-factor method and orthogonal test.[Results]The bacteria strain was identification as bacillus（Bacillus sp.） with the traditional method and molecular biological method.With soluble starch,corn flour or wheat flour as a carbon source,the strain-B-5 contributed to enzyme-producing.From the viewpoint of the cost reduction in industrial production,the corn flour could be adopted as a carbon source in its fermentation.And the tryptone was selected as nitrogen source because the highest activity of enzyme in its fermentation was produced by the nitrogen source.The orthogonal experiment results showed that it was most conducive to fermentation when the component of corn flour and tryptone in medium was 2%,respectively,with the activity of enzyme of 74.6 U/ml.The highest activity of enzyme in the B-5 fermentation appeared at the treatment time of 60th hour and the activity of enzyme was gradually decreased with time-lasting.[Conclusion]The activity of the enzyme of strain-B-5 was still limited,which yet could not meet the needs of industrial production,although nature of the acidic α-amylase produced by the strain was excellent.It was beneficial to the production of the bacteria strain of acidic α-amylase-producing for its industrialization through the identification of the gene in the strain.

关 键 词：酸性Α-淀粉酶 菌种鉴定 芽孢杆菌 发酵条件 

分 类 号：S182[农业科学—农业基础科学]