Cis-Element Engineering Promotes the Expression of Bacillus subtilis Type I L-Asparaginase and Its Application in Food
Cis-Element Engineering Promotes the Expression of Bacillus subtilis Type I L-Asparaginase and Its Application in Food
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顺式元件工程促进枯草芽孢杆菌I型L-天冬酰胺酶的表达及其在食品中的应用
DOI：
10.3390/ijms23126588
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发表时间：
2022-06-13
影响因子：
5.6
通讯作者：
Lu, Fengxia
中科院分区：
文献类型：
--
作者：
Niu, Jiafeng;Yan, Ruxue;Shen, Juan;Zhu, Xiaoyu;Meng, Fanqiang;Lu, Zhaoxin;Lu, Fengxia
关键词：
地衣芽孢杆菌（Bacilluslicheniformis）Z-1 Ⅰ型L-天冬酰胺酶（BlAase）在枯草芽孢杆菌（Bacillussubtilis）RIK 1285中高效分泌，但产量较低，不适合工业化生产。因此，在本研究中使用组合方法来促进B中的BlAase合成。枯草杆菌。首先，选择15个单一强启动子来替换原始启动子P43，其中PyvyD实现最大BlAase活性（436.28U/mL）。第二，使用具有相对高的BlAase表达水平的四个启动子（PyvyD、P43、PaprE和PspoVG）构建双启动子系统以提高BlAase产量，其中启动子PaprE-PyvyD的引擎达到502.11 U/mL。通过修饰PaprE-PyvyD启动子的关键部分，BlAase的活性也增加（568.59U/mL）。第三，当启动子PyvyD的核糖体结合位点（RBS）序列被替换时，BlAase活性达到790.1U/mL，是原始启动子P43菌株的2.27倍。在10 L发酵罐中培养36 h后，BlAase的表达量达到2163.09 U/mL，是使用启动子P43的出发菌株的6.2倍。此外，还对BlAase在马铃薯片中丙烯酰胺迁移方面的应用潜力进行了评价。结果表明，采用热烫和BlAase联合处理，油炸薯片中丙烯酰胺的去除率可达89.50%。这些研究结果表明，结合转录和翻译技术是提高重组蛋白产量的有效策略，BlAase可以作为控制食品加工中丙烯酰胺的一个很好的候选者。
Type I L-asparaginase from Bacillus licheniformis Z-1 (BlAase) was efficiently produced and secreted in Bacillus subtilis RIK 1285, but its low yield made it unsuitable for industrial use. Thus, a combined method was used in this study to boost BlAase synthesis in B. subtilis. First, fifteen single strong promoters were chosen to replace the original promoter P43, with PyvyD achieving the greatest BlAase activity (436.28 U/mL). Second, dual-promoter systems were built using four promoters (PyvyD, P43, PaprE, and PspoVG) with relatively high BlAase expression levels to boost BlAase output, with the engine of promoter PaprE-PyvyD reaching 502.11 U/mL. The activity of BlAase was also increased (568.59 U/mL) by modifying key portions of the PaprE-PyvyD promoter. Third, when the ribosome binding site (RBS) sequence of promoter PyvyD was replaced, BlAase activity reached 790.1 U/mL, which was 2.27 times greater than the original promoter P43 strain. After 36 h of cultivation, the BlAase expression level in a 10 L fermenter reached 2163.09 U/mL, which was 6.2 times greater than the initial strain using promoter P43. Moreover, the application potential of BlAase on acrylamide migration in potato chips was evaluated. Results showed that 89.50% of acrylamide in fried potato chips could be removed when combined with blanching and BlAase treatment. These findings revealed that combining transcription and translation techniques are effective strategies to boost recombinant protein output, and BlAase can be a great candidate for controlling acrylamide in food processing.
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影响因子：
6.1
作者：
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通讯作者：
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影响因子：
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