Characterization of a Novel L-Asparaginase from Mycobacterium gordonae with Acrylamide Mitigation Potential.
Characterization of a Novel L-Asparaginase from Mycobacterium gordonae with Acrylamide Mitigation Potential.
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具有丙烯酰胺缓解潜力的戈登分枝杆菌新型 L-天冬酰胺酶的表征
DOI：
10.3390/foods10112819
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发表时间：
2021-11-16
期刊：
影响因子：
--
通讯作者：
Lu F
中科院分区：
文献类型：
--
作者：
Chi H;Chen M;Jiao L;Lu Z;Bie X;Zhao H;Lu F
L-天冬酰胺酶（E.C.3.5.1.1）是一种众所周知的药物，可在食品工业中防止丙烯酰胺的形成，并在临床环境中预防儿童急性淋巴细胞白血病。L-天冬酰胺酶的pH稳定范围窄、热稳定性差等缺点限制了其工业应用。本研究从戈登分枝杆菌中克隆了一种新的L-天冬酰胺酶（GmASNase），并在大肠杆菌BL 21（DE 3）中表达。GmASNase是一个单体大小为32 kDa的四聚体，与蛋白质数据库中的幽门螺杆菌L-天冬酰胺酶只有32%的结构相同。纯化的GmASNase在pH 9.0和50 °C下具有最高的比活性，为486.65 IU mg−1。此外，GmASNase在5.0-11.0的宽pH范围内的稳定性和在低于40 °C的温度下的活性方面具有优越的上级性质。此外，GmASNase对L-天冬酰胺显示出高底物特异性，Km、kcat和kcat/Km值分别为6.025 mM、11，864.71 min−1和1969.25 mM− 1 min −1。为了评估其减轻丙烯酰胺的能力，GmASNase用于在油炸之前处理薯片，其中与未处理的对照相比，丙烯酰胺含量降低了65.09%。这些结果表明，GmASNase是食品工业中应用的潜在候选者。
L-asparaginase (E.C.3.5.1.1) is a well-known agent that prevents the formation of acrylamide both in the food industry and against childhood acute lymphoblastic leukemia in clinical settings. The disadvantages of L-asparaginase, which restrict its industrial application, include its narrow range of pH stability and low thermostability. In this study, a novel L-asparaginase from Mycobacterium gordonae (GmASNase) was cloned and expressed in Escherichia coli BL21 (DE3). GmASNase was found to be a tetramer with a monomeric size of 32 kDa, sharing only 32% structural identity with Helicobacter pylori L-asparaginases in the Protein Data Bank database. The purified GmASNase had the highest specific activity of 486.65 IU mg−1 at pH 9.0 and 50 °C. In addition, GmASNase possessed superior properties in terms of stability at a wide pH range of 5.0–11.0 and activity at temperatures below 40 °C. Moreover, GmASNase displayed high substrate specificity towards L-asparagine with Km, kcat, and kcat/Km values of 6.025 mM, 11,864.71 min−1 and 1969.25 mM−1min−1, respectively. To evaluate its ability to mitigate acrylamide, GmASNase was used to treat potato chips prior to frying, where the acrylamide content decreased by 65.09% compared with the untreated control. These results suggest that GmASNase is a potential candidate for applications in the food industry.
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DOI：
10.1016/j.ijbiomac.2020.01.012
发表时间：
2020-03-15
影响因子：
8.2
作者：
Chohan, Shahid Mahmood;Sajed, Muhammad;Rashid, Naeem
通讯作者：
Rashid, Naeem
影响因子：
0.8
作者：
Hassan, Sahar W. M.;Farag, Aida M.;Beltagy, Ehab A.
通讯作者：
Beltagy, Ehab A.
影响因子：
2.8
作者：
Onishi Y;Prihanto AA;Yano S;Takagi K;Umekawa M;Wakayama M
通讯作者：
Wakayama M
影响因子：
4.4
作者：
Kumar, Sanjay;Dasu, Veeranki Venkata;Pakshirajan, Kannan
通讯作者：
Pakshirajan, Kannan
影响因子：
2.9
作者：
Dash, Chitrangada;Mohapatra, Sukanti Bala;Maiti, Prasanta Kumar
通讯作者：
Maiti, Prasanta Kumar