Novel extremophilic proteases from Pseudomonas aeruginosa M211 and their application in the hydrolysis of dried distiller's grain with solubles
Novel extremophilic proteases from Pseudomonas aeruginosa M211 and their application in the hydrolysis of dried distiller's grain with solubles
复制标题
铜绿假单胞菌 M211 新型嗜极蛋白酶及其在干酒糟可溶物水解中的应用
DOI：
10.1002/btpr.2728
复制
发表时间：
2018
影响因子：
2.9
通讯作者：
Flores-Fernández C
中科院分区：
文献类型：
--
作者：
Flores-Fernández C
蛋白水解酶是最重要的一类工业酶，可用于多种领域，包括用于工业副产品价态的生物精炼。在这项研究中，我们纯化并鉴定了从秘鲁亚马逊沼泽土壤样品中分离到的一株铜绿假单胞菌产生的新型嗜极端蛋白酶。此外，我们还测试了它们对酒糟可溶干蛋白(DDGS)的水解力。纯化了三种碱性和嗜热性丝氨酸蛋白酶EI、EII和EIII，它们的相对分子质量分别为35、40和55 kDa。EI和EIII被EDTA和Pefabloc强烈抑制，被归类为丝氨酸金属蛋白酶，而EII仅被Pefabloc完全抑制，被归类为丝氨酸蛋白酶。此外，EI和EII在pH=8时酶活最高，而EIII酶活性在pH 11时几乎保持100%，在pH 12时酶活几乎保持在100%。在60℃时，所有酶的活性都以60℃最高。用纯化的铜绿假单胞菌M211酶对DDGS蛋白进行了水解，结果表明，基于释放的甘氨酸，EIII对DDGS具有最高的蛋白分解活性。该酶可释放小麦DDGS蛋白中总甘氨酸含量的63%，是使用商用Pronase®时的2.2倍。综上所述，我们的结果表明这种新型的酶在DDGS的水解中有很大的应用潜力。2018年美国化学工程师学会生物技术。程序，35：E2728,2019年
Proteases are the most important group of industrial enzymes and they can be used in several fields including biorefineries for the valorization of industrial byproducts. In this study, we purified and characterized novel extremophilic proteases produced by aPseudomonas aeruginosastrain isolated fromMauritia flexuosapalm swamps soil samples in Peruvian Amazon. In addition, we tested their ability to hydrolyze distillers dried grains with solubles (DDGS) protein. Three alkaline and thermophilic serine proteases named EI, EII, and EIII with molecular weight of 35, 40, and 55 kDa, respectively, were purified. EI and EIII were strongly inhibited by EDTA and Pefabloc being classified as serine‐metalloproteases, while EII was completely inhibited only by Pefabloc being classified as a serine protease. In addition, EI and EII exhibited highest enzymatic activity at pH 8, while EIII at pH 11 maintaining almost 100% of it at pH 12. All the enzymes demonstrated optimum activity at 60°C. Enzymatic activity of EI was strongly stimulated in presence of Mn2+(6.9‐fold), EII was stimulated by Mn2+(3.7‐fold), while EIII was slightly stimulated by Zn2+, Ca2+, and Mg2+. DDGS protein hydrolysis using purifiedPseudomonas aeruginosaM211 proteases demonstrated that, based on glycine released, EIII presented the highest proteolytic activity toward DDGS. This enzyme enabled the release 63% of the total glycine content in wheat DDGS protein, 2.2‐fold higher that when using the commercial Pronase®. Overall, our results indicate that this novel extremopreoteases have a great potential to be applied in DDGS hydrolysis. © 2018 American Institute of Chemical EngineersBiotechnol. Prog., 35: e2728, 2019
登录
查看更多内容
影响因子：
2.8
作者：
Karadzic, I;Masui, A;Fujiwara, N
通讯作者：
Fujiwara, N
东加尔各答湿地铜绿假单胞菌胞外蛋白酶的纯化和表征
DOI：
--
发表时间：
2010
期刊：
影响因子：
--
作者：
J. S. Yadav;Sanhita Chowdhury;S. Chaudhuri
通讯作者：
S. Chaudhuri
影响因子：
2.8
作者：
Ibrahim, Kalibulla Syed;Muniyandi, Jeyaraj;Pandian, Shunmugiah Karutha
通讯作者：
Pandian, Shunmugiah Karutha
来自假单胞菌属的胞外蛋白酶。
影响因子：
4.4
作者：
S. Shastry;M. Prasad
通讯作者：
M. Prasad
皮革和洗涤剂行业中的新型微生物蛋白酶。
DOI：
--
发表时间：
2013
期刊：
影响因子：
--
作者：
F. Khan
通讯作者：
F. Khan