Screening, identification and growth characteristics of a low-temperature proteinase producing strain
-
摘要: 以新疆阿勒泰地区的冻土为原材料,采用平板分离法初筛,测蛋白酶活性法复筛,筛选出9株低温蛋白酶产生菌株,其中菌株A29-2蛋白酶活性最高,为31.88U/mL,且该菌在初始pH 7.0,NaCl质量分数为1.00%,22℃条件下繁殖量最大。通过形态学观察、生理生化鉴定及16S rDNA序列鉴定,鉴定为:该菌属于沙雷氏菌(Serratia sp.),该菌株所产的蛋白酶在4℃条件下保存48h,酶活力基本保持不变,表明该低温耐冷菌所产的蛋白酶为低温酶。Abstract: Through the plate separation and the proteinase activity determination, nine low- temperature proteinase producing strains were obtained from the permafrost in the region of Altai, Xinjiang, in which the strain A29-2 had the highest proteinase activity, reaching to 31.88U /mL.The growth of A29-2 reached the maximum level under the condition of initial pH7.0, mass fraction of NaCl 1.00%, temperature 22℃.Based on the morphological observation, physiological and biochemical identification and 16S rDNA sequence analysis, the strain A29- 2 was identified as Serratia sp..The activity of proteinase produced by A29-2 remained mostly unchanged after 48h of stored at 4℃.The results indicated that proteinase produced by cold-adapted strains was low-temperature enzyme.
-
[1] Morita R.Y.Psychrophilic bacteria[J].Bacterial Rev, 1975, 39:144-167.
[2] Gerday C, Aittaleb M, Bentahir M, et al.Cold-adapted enzymes:from fundamentals to biotechnology[J].Trends in Biotechnol, 2000, 18 (3) :103-107.
[3] 王长远, 罗梦晓.高产蛋白酶地衣芽孢杆菌株的紫外诱变[J].中国生物制品学杂志, 2012, 25 (5) :620-622. [4] 祁丹丹, 杨瑞金, 华霄, 等.天山冻土中产低温蛋白酶菌株的筛选、鉴定及酶学性质[J].食品与发酵工业, 2012, 38 (7) :32-37. [5] Vazquez SC, Coria SH, Cormack WPM.Extracellular proteases from eight psychrotolerant antarctic strains[J].Microbiol Res, 2004, 159 (2) :157-166.
[6] 陈秀兰, 张玉忠, 高培基.低温蛋白酶的研究进展与应用前景[J].工业微生物, 2001, 31 (1) :52-57. [7] Margesin R, Dieplinger H, Hofmann J, et al.A cold-active extracellular metalloprotease from Pedobacter cryoconitis:production and properties[J].Research in Microbiology, 2005, 156 (4) :499-505.
[8] 黄秀梨, 辛明秀.微生物学实验指导[M].北京:高等教育出版社, 2008:19-23. [9] 魏群.分子生物学实验指导[M].北京:高等教育出版社, 2007:25-26. [10] 周德庆.微生物学实验教程[M].北京:高等教育出版社, 2006:49-51, 220-225. [11] 东秀珠, 蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社, 2001:66-95. [12] Buchanan R E, Gibbens N E.伯杰细菌鉴定手册 (第八版) [M].北京:科学出版社, 1984:450-454. [13] 李远, 巴吐尔, 张小燕, 等.新疆哈萨克族传统发酵驼乳中乳酸菌的分离鉴定[J].中国食物与营养, 2011, 17 (1) :54-58. [14] 马桂珍, 暴增海, 王淑芳, 等.高产蛋白酶细菌的分离筛选及其种类鉴定[J].食品科学, 2011, 32 (21) :183-187. [15] 迟乃玉, 张庆芳, 王晓辉, 等.海洋低温蛋白酶菌株选育及发酵培养基的研究 (I) [J].微生物学通报, 2006, 33 (1) :114-117. [16] 刘静, 闵航, 章骥, 等.一株产低温蛋白酶菌株的筛选鉴定及纯酶研究[J].浙江大学学报, 2006, 32 (3) :251-256. [17] 全桂静, 尹黎献.低温碱性蛋白酶高产菌株的选育与发酵条件研究[J].沈阳化工大学学报, 2011, 25 (2) :117-120.
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量: