定点突变提高食品新型L-天冬酰胺酶的活力及热稳定性

doi:10.3969/j.issn.1673-1689.2020.09.014

首页 > 过刊浏览>2020年第39卷第9期 >99-104. DOI:10.3969/j.issn.1673-1689.2020.09.014

定点突变提高食品新型L-天冬酰胺酶的活力及热稳定性
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                        10.3969/j.issn.1673-1689.2020.09.014
                    
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Improving the Activity and Thermal Stability of Pyrococcus yayanosii CH1 L-Asparaginase by Site-Directed Mutagenesis

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摘要:

L-天冬酰胺酶（L-asparaginase II，EC 3.5.1.1）可将L-天冬酰胺转化为L-天冬氨酸，减少高温加工食品中丙烯酰胺的形成，因而受到人们的广泛关注。该酶在食品加工及预处理阶段的使用已受到人们广泛的关注，但是由于食品加工及预处理过程中环境的复杂性，对L-天冬酰胺酶的性质、热稳定性和酶活等方面有较高的要求。通过序列对比和同源模拟对嗜热菌Pyrococcus yayanosii CH1来源的编码L-天冬酰胺酶的基因PyAsnase进行了3个位点的突变，并在Bacilus subtilis 168 中进行表达，提高了该酶的热稳定性及比酶活。其中突变株E22K较原始菌株相比所得突变体比酶活提高了约37.3%，突变株R111L较原始菌株相比所得突变体的比酶活提高了约31.1%，突变株M92A较原始突变菌株相比所得突变体在85 ℃时的半衰期延长了约30 min。本研究结果为探索L-天冬酰胺酶结构和功能的相互关系提供了借鉴，提高了其在食品工业中的应用前景。

Abstract:

L-asparaginase II (EC 3.5.1.1), which can convert L-asparagine, the precursor of carcinogenic acrylamide, to L-aspartic acid, is attracting much attention in food industry. The use of L-asparaginase II can reduce the formation of acrylamide during the pretreatment of foods. However, the application of this enzyme is limited due to variant requirements of commercial use, especially for its activity and thermostability. In this study, by sequence aligning and homology modeling,three mutants of PyAsnase gene from the thermophilus Pyrococcus yayanosii CH1was constructed and over-expressed in Bacillus subtilis. According to the results, the specific activities of the mutants E22K and R111L were increased by 37.3% and 31.1%, respectively, and the half-life of the mutant obtained by the mutant M92A was extended by 30 min at 85 ℃. This work provides references for studying the interrelationship between function and structure of L-asparaginase, and also develops the application of L-asparaginase in food industry.

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徐书琴,胡静怡,张恒维,赵浩东,刘鹏,席毓淋,李谞,张显,饶志明.定点突变提高食品新型L-天冬酰胺酶的活力及热稳定性[J].食品与生物技术学报,2020,39(9):99-104.

XU Shuqin, HU Jingyi, ZHANG Hengwei, ZHAO Haodong, LIU Peng, XI Yulin, LI Xu, ZHANG Xian, RAO Zhiming. Improving the Activity and Thermal Stability of Pyrococcus yayanosii CH1 L-Asparaginase by Site-Directed Mutagenesis[J]. Journal of Food Science and Biotechnology,2020,39(9):99-104.

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在线发布日期: 2020-11-26
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