Engineering of Yersinia Phytases to Improve Pepsin and Trypsin Resistance and Thermostability and Application Potential in the Food and Feed Industry

Niu Canfang; Yang Peilong; Luo Huiying; Huang Huoqing; Wang Yaru; Yao Bin

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Engineering of Yersinia Phytases to Improve Pepsin and Trypsin Resistance and Thermostability and Application Potential in the Food and Feed Industry

机译：益苯浮糖酶的工程改善食品和饲料工业中的胃蛋白和胰蛋白酶抗性和胰蛋白酶抵抗力和应用潜力

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Susceptibility to proteases usually limits the application of phytase. We sought to improve the pepsin and trypsin resistance of YeAPPA from Yersinia enterocolitica and YkAPPA from Y. kristensenii by optimizing amino acid polarity and charge. The predicted pepsin/trypsin cleavage sites F89/K226 in pepsin/trypsin-sensitive YeAPPA and the corresponding sites (F89/E226) in pepsin-sensitive but trypsin-resistant YkAPPA were substituted with S and H, respectively. Six variants were produced in Pichia pastoris for catalytic and biochemical characterization. F89S, E226H, and F89S/E226H elevated pepsin resistance and thermostability and K226H and F89S/K226H improved pepsin and trypsin resistance and stability at 60 degrees C and low pH. All the variants increased the ability of the proteins to hydrolyze phytate in corn meal by 2.6-14.9-fold in the presence of pepsin at 37 degrees C and low pH. This study developed a genetic manipulation strategy specific for pepsin/trypsin-sensitive phytases that can improve enzyme tolerance against proteases and heat and benefit the food and feed industry in a cost-effective way.

机译：对蛋白酶的易感性通常限制植酸酶的应用。我们试图通过优化氨基酸极性和电荷，改善来自Yersinia Enterocolitica和Ykappa的YeAppa的胃蛋白酶和胰蛋白酶抵抗力。百事可纲/胰蛋白酶敏感性Yeappa中预测的百草蛋白/胰蛋白酶切割位点F89 / K226和胃蛋白酶敏感性但胰蛋白酶抗性Ykappa的相应部位（F89 / E226）分别被S和H取代。在Pichia Pastoris中产生了六种变体，用于催化和生化表征。 F89S，E226H和F89S / E226H高胃素抗性和热稳定性，K226H和F89S / K226H改善胃蛋白酶和胰蛋白酶抗性，稳定性在60℃和低pH下。所有变体都会增加蛋白质在玉米粉中水解植物水解的能力2.6-14.9倍，在胃蛋白蛋白在37℃和低pH下。本研究开发了针对胃蛋白酶/胰蛋白酶敏感植物酶特异性的遗传操作策略，可提高蛋白酶和热量的酶耐受，并以成本效益的方式使食品和饲料行业受益。

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来源
《Journal of Agricultural and Food Chemistry》 |2017年第34期|共8页
作者
Niu Canfang; Yang Peilong; Luo Huiying; Huang Huoqing; Wang Yaru; Yao Bin;
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作者单位

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

Chinese Acad Agr Sci Feed Res Inst Minist Agr Natl Engn Res Ctr Biol Feed Key Lab Feed Biotechn 12 Zhongguancun South St Beijing 100081 Peoples R China;

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原文格式 PDF
正文语种 eng
中图分类营养卫生、食品卫生;农业科学;
关键词
phytases; pepsin resistance; trypsin resistance; thermostability; stability at low pH; hydrolysis of corn meal phytate; optimizing the polarity and charge of the amino acids;

机译：Phytase;胃蛋白酶抗性;胰蛋白酶抗性;热稳定性;低pH值稳定性;水解玉米粉植物;优化氨基酸的极性和电荷;

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专利

1. Engineering of Yersinia Phytases to Improve Pepsin and Trypsin Resistance and Thermostability and Application Potential in the Food and Feed Industry [J] . Niu Canfang, Yang Peilong, Luo Huiying, Journal of Agricultural and Food Chemistry . 2017,第34期

机译：益苯浮糖酶的工程改善食品和饲料工业中的胃蛋白和胰蛋白酶抗性和胰蛋白酶抵抗力和应用潜力
2. N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites [J] . Canfang Niu, Huiying Luo, Pengjun Shi, Applied and Environmental Microbiology . 2016,第4期

机译：N-糖基化可增强组氨酸酸性磷酸酶磷酸酶的胃蛋白酶抵抗力，方法是增强其在酸性pH值下的稳定性，并降低胃蛋白酶对其裂解位点的可及性。
3. N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites [J] . Canfang Niu, Huiying Luo, Pengjun Shi, Applied Microbiology . 2016,第4期

机译：N-糖基化可增强组氨酸酸性磷酸酶磷酸酶的胃蛋白酶抵抗力，方法是增强其在酸性pH值下的稳定性，并降低胃蛋白酶对其裂解位点的可及性。
4. Thermostable xylanases and their potential application in paper and pulp industries [C] . M K Bhat, S Kalogiannis, N A Bennett, Cellucon conference . 2000

机译：热稳定的木聚糖酶及其在纸张和纸浆行业的潜在应用
5. Rapid Quantification of Trypsin Inhibitors in Food and Feed Formulations by Electrospray Ionization Mass Spectrometry. [D] . Panta, Radheshyam. 2017

机译：通过电喷雾电离质谱快速定量食品和饲料配方中的胰蛋白酶抑制剂。
6. Engineering the residual side chains of HAP phytases to improve their pepsin resistance and catalytic efficiency [O] . Canfang Niu, Peilong Yang, Huiying Luo, -1

机译：工程化HAP植酸酶的残留侧链以提高其胃蛋白酶抵抗力和催化效率
7. N -Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites [O] . Canfang Niu, Huiying Luo, Pengjun Shi, 2016

机译：N-Glycosylation通过提高酸性pHS的稳定性并降低胃蛋白酶对其切割位点的可接受来改善组氨酸酸性磷酸酶植物酶的胃蛋白酶抗性
8. Hazard Evaluation Division, Standard Evaluation Procedure: Specialty Applications. 1. Classification of Seed Treatments and Treatment of Crops Grown for Seed Use Only as Nonfood or Food Uses. 2. Magnitude of the Residue: Postharvest Fumigation of Crops and Processed Foods and Feeds. 3. Magnitude of the Residue: Postharvest Treatment (Except Fumigation) of Crops and Processed Foods and Feeds [R] . Loftus, M. L. , Metzger, M. S. 1988

机译：危害评估部门，标准评估程序：专业应用。 1.种子处理的分类和种子作为非食品或食品用途种植的作物的处理。 2.残留物的大小：作物和加工食品和饲料的采后熏蒸。 3.残留物的大小：作物和加工食品和饲料的采后处理（熏蒸除外）

Engineering of Yersinia Phytases to Improve Pepsin and Trypsin Resistance and Thermostability and Application Potential in the Food and Feed Industry

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