Study on isolation and identification of acidic α-amylase producing strain and enzymatic properties
-
摘要: 用含有淀粉的选择培养基,通过透明圈法筛选到一株产酸性淀粉酶的菌株,其发酵液酶活力达到123U/mL。通过形态观察。生理生化实验和16S rDNA序列分析等方法初步鉴定该菌株为地衣芽孢杆菌,命名为Bacillus lincheniformis BW-2。对其产生的淀粉酶的酶学性质进行分析发现,该菌株产生的淀粉酶最适反应温度为70℃;最适反应pH为4.5,同时具有较好的热稳定性和pH稳定性;Ca2+和Mg2+对该酶有激活作用,Mn2+和Cu2+对该酶有抑制作用,抑制剂PMSF（苯甲基磺酰氟）及DTT（二硫苏糖醇）对该酶活力影响较小。通过PAGE分析发现BW-2产生的淀粉酶与已知热稳定淀粉酶分子量或结构可能存在差异。Bacillus lincheniformis BW-2是一株具有研究开发潜力的产酸性淀粉酶的菌株。Abstract: An acidic α-amylase producing strain was screened through screening medium supplemented with starch by the method of hydrolysis halo, and the amylase activity of the fermentation broth was 123U/mL. The stain was identified as Bacillus lincheniformis by morphology observation, physiological and biochemical tests and 16S rDNA sequence analysis, and was named as Bacillus lincheniformis BW-2. The enzymatic properties of the amylase were also investigated. The optimum pH and temperature of the amylase was 4.5 and 70℃, respectively. It also showed excellent thermostability and pH stability. The amylase activity was activated by Ca2+and Mg2+, whereas, it was inhibited by Mn2+and Cu2+. Meanwhile, these inhibitors PMSF and DTT had little effect on the enzyme activity. The molecular weight and structure of the amylase from BW-2 maybe was different from the known thermostable amylase by PAGE analysis. BW-2 was an acidic amylase producing strain which showed research and development potential.
-
Keywords:
- acidic amylase /
- screening /
- isolation and identification /
- enzymatic properties
-
[1] Rana N, Walia A, Gaur A.α-Amylases from microbialsources and its potential applications in various industries[J].National Academy Science Letters, 2013, 36 (1) :9-17.
[2] Maarel V D M, Bart V D V, Joost C M, et al.Properties and applications of starch converting enzymes of the a-amylase family[J].Journal of Biotechnology, 2002, 94:137-155.
[3] 刘旭东, 徐岩.一种新的中温酸性α-淀粉酶的分离纯化及酶学性质[J].应用与环境微生物学报, 2008, 14 (2) :235-239. [4] 王晓燕, 高润池, 廖昌珑, 等.Geobacillus sp.A27的分离筛选及其淀粉酶酶学性质研究[J].食品工业科技, 2011, 32 (1) :151-155. [5] Pandey A, Nigam P, Soccol C R, et al.Advences in microbial amylases[J].Biotechnology and Applied Biochemistry, 31 (Pt2) , 2000:135-152.
[6] Sivaramakrishnan S, Gangadharan D, Nampoothiri K M, et al.Alpha-amylases from microbial sources-an overview on recent developments[J].Food Technology and Biotechnology, 2006, 44 (2) :173-184.
[7] Sajedi R H, Naderi-Manesh H, Khajeh K, et al.A Caindependent alpha-amylase that is active and stable at low pH from the Bacillus sp.KR-8104[J].Enzyme and Microbial Technology, 2005, 36 (5-6) :666-671.
[8] Goyal N, Gupta J K, Soni S K.A novel raw starch digesting thermostableα-amylase from Bacillus sp.I-3 and its use in the direct hydrolysis of raw potato starch[J].Enzyme and Microbial Technology, 2005, 37:723-734.
[9] Gupta R, Gigras P, Mohapatra H, et al.Microbialα-amylase:a biotechnological perspective[J].Process Biochemistry, 2003, 38:1599-1616.
[10] Crab W, Mitchinson C.Enzymes involved in the processing of starch to sugars[J].Trends Biotechnology, 1997, 15:349-352.
[11] Shoji H, Toshikazu S, Kenji H, et al.Simultaneous production of glucoamylase and acid-stableα-amylase using novel submerged culture of Aspergillus kawachii NBRC4308[J].Jounal of Bioscience and Bioengineering, 2007, 103:203-205.
[12] 谢建华, 师永生, 杜丽琴, 等.一株产酸性α-淀粉酶菌株的筛选、纯化及酶学性质[J].应用与环境生物学报, 2011, 17 (1) :95-99. [13] 钱萍.酸性淀粉酶菌株的诱变选育及酶学性质研究[J].食品工业科技, 2011, 33 (11) :183-186. [14] 冯旭明, 迟乃玉, 张庆芳.低温淀粉酶菌株C2的分离鉴定及其产低温淀粉酶的酶学性质[J].微生物学通报, 2011, 38 (12) :1762-1767. [15] 刘洋, 唐芳荣, 相宏宇, 等.小体系高通量比色测定糖化酶活力新方法[J].中国酿造, 2012, 31 (7) :140-143. [16] Hot G J, Krieg R N, Sneath HAP, et al.Bergey’s Manual of Determinative Bacteriology[M].9th ed.Baltimore:Williams and Wilkins, 1994.
[17] 张保国, 唐玲李祖明, 等.阿维菌素杀虫剂对甘蓝叶际微生物群落结构的影响[J].环境科学, 2009, 30 (5) :1291-1297. [18] 秦臻, 蔡素梅, 黄钧, 等.一株产生淀粉分解酶犁头霉的分离鉴定及其酶学性质[J].微生物学通报, 2011, 38 (5) :729-735. [19] Burhan Arikan.Highly thermostable, thermophilic, alkaline, SDS and chelator resistant amylase from a thermophilic Bacillus sp.isolate A3-15[J].Bioresource Technology, 2008, 99:3071-3076.
[20] Xu D L, Yan X.A novel raw starch digestingα-amylase from a newly isolated Bacillus sp.YX-1:Purification and characterization[J].Bioresource Technology, 2008, 99:4315-4320.
计量
- 文章访问数: 128
- HTML全文浏览量: 8
- PDF下载量: 409
下载:
