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膜生物反应器中从脱脂豆粕生产低敏抗菌肽:一项具有全面产物表征的生物过程工程研究

Production of Hypoallergenic Antibacterial Peptides from Defatted Soybean Meal in Membrane Bioreactor: A Bioprocess Engineering Study with Comprehensive Product Characterization.

作者信息

Nath Arijit, Szécsi Gábor, Csehi Barbara, Mednyánszky Zsuzsa, Kiskó Gabriella, Bányai Éva, Dernovics Mihály, Koris András

机构信息

Department of Food Engineering, Faculty of Food Science, Szent István University, Ménesi st 44, 
HU-1118 Budapest, Hungary.

Department of Refrigeration and Livestock Product Technology, Faculty of Food Science, 
Szent István University, Ménesi st 43-45, HU-1118 Budapest, Hungary.

出版信息

Food Technol Biotechnol. 2017 Sep;55(3):308-324. doi: 10.17113/ftb.55.03.17.5040.

Abstract

Hypoallergenic antibacterial low-molecular-mass peptides were produced from defatted soybean meal in a membrane bioreactor. In the first step, soybean meal proteins were digested with trypsin in the bioreactor, operated in batch mode. For the tryptic digestion of soybean meal protein, optimum initial soybean meal concentration of 75 g/L, temperature of 40 °C and pH=9.0 were determined. After enzymatic digestion, low-molecular-mass peptides were purified with cross-flow flat sheet membrane (pore size 100 µm) and then with tubular ceramic ultrafiltration membrane (molecular mass cut-off 5 kDa). Effects of transmembrane pressure and the use of a static turbulence promoter to reduce the concentration polarization near the ultrafiltration membrane surface were examined and their positive effects were proven. For the filtration with ultrafiltration membrane, transmembrane pressure of 3·10 Pa with 3-stage discontinuous diafiltration was found optimal. The molecular mass distribution of purified peptides using ultrafiltration membrane was determined by a liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry setup. More than 96% of the peptides (calculated as relative frequency) from the ultrafiltration membrane permeate had the molecular mass ≤1.7 kDa and the highest molecular mass was found to be 3.1 kDa. The decrease of allergenic property due to the tryptic digestion and membrane filtration was determined by an enzyme-linked immunosorbent assay and it was found to exceed 99.9%. It was also found that the peptides purified in the ultrafiltration membrane promoted the growth of HA6111-2 and they possessed antibacterial activity against .

摘要

在膜生物反应器中由脱脂豆粕制备低变应原性抗菌低分子量肽。第一步,在以分批模式运行的生物反应器中用胰蛋白酶消化豆粕蛋白。对于豆粕蛋白的胰蛋白酶消化,确定了最佳初始豆粕浓度为75 g/L、温度为40°C和pH = 9.0。酶消化后,先用错流平板膜(孔径100 µm)纯化低分子量肽,然后用管状陶瓷超滤膜(截留分子量5 kDa)纯化。研究了跨膜压力和使用静态湍流促进剂以降低超滤膜表面附近浓差极化的影响,并证明了它们的积极作用。对于超滤膜过滤,发现3×10 Pa的跨膜压力和三级间断渗滤是最佳的。使用液相色谱 - 电喷雾电离四极杆飞行时间质谱装置测定了用超滤膜纯化的肽的分子量分布。超滤膜渗透物中超过96%的肽(以相对频率计算)分子量≤1.7 kDa,发现最高分子量为3.1 kDa。通过酶联免疫吸附测定法确定了由于胰蛋白酶消化和膜过滤导致的变应原性降低,发现其超过99.9%。还发现超滤膜中纯化的肽促进了HA6111 - 2的生长,并且它们对……具有抗菌活性。

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本文引用的文献

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Role of lipids in the interaction of antimicrobial peptides with membranes.脂质在抗菌肽与膜相互作用中的作用。
Prog Lipid Res. 2012 Apr;51(2):149-77. doi: 10.1016/j.plipres.2011.12.005. Epub 2012 Jan 8.

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